Progress in Neurobiology 83 (2007) 110–130

www.elsevier.com/locate/pneurobio

The discovery of dendritic spines by Cajal in 1888 and its relevance

in the present neuroscience
Pablo Garcı́a-López, Virginia Garcı́a-Marı́n *, Miguel Freire
Museo Cajal, Instituto Cajal, CSIC, Avda. Doctor Arce 37, 28002 Madrid, Spain
Received 27 October 2006; received in revised form 17 February 2007; accepted 3 April 2007

Abstract
The year 2006 marks the centenary of the Nobel Prize for Physiology or Medicine awarded to Santiago Ramón y Cajal and Camilo Golgi, ‘‘in
recognition of their work on the structure of the nervous system’’. Their discoveries are keys to understanding the present neuroscience, for
instance, the discovery of dendritic spines. Cajal discovered dendritic spines in 1888 with the Golgi method, although other contemporary scientists
thought that they were silver precipitates. Dendritic spines were demonstrated definitively as real structures by Cajal with the Methylene Blue in
1896. Many of the observations of Cajal and other contemporary scientists about dendritic spines are active fields of research of present
neuroscience, for instance, their morphology, distribution, density, development and function. This article will deal with the main contributions of
Cajal and other contemporary scientists about dendritic spines. We will analyse their contributions from the historical and present point of view. In
addition, we will show high quality images of Cajal’s original preparations and drawings related with this discovery.
# 2007 Elsevier Ltd. All rights reserved.

Keywords: Cajal; Dendritic spines; Filopodia; Golgi method; Methylene Blue method

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
2. The historical context of the discovery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
3. Principles of Cajal’s scientific reasoning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111
4. Principal data contributed by Cajal to the research of the dendritic spines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
4.1. Morphological data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
4.2. Distribution of spines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115
4.3. Development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 117
4.4. Physiological role of the dendritic spines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
4.5. Dendritic spines in pathological and poisoning states . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
5. The dendritic spines from Cajal to present-day neuroscience. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5.1. The concept of Sherrington’s synapse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
5.2. Morphology of dendritic spines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.3. Dendritic spines and pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
5.4. Plasticity of dendritic spines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124
5.5. Dendritic spines and development . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 125
6. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127

Abbreviations: s.s., sensu strictus; CA1, cornu ammonis 1; CA3, cornu ammonis 3; EM, electron microscopy; ER, endoplasmic reticulum; LTP, long-term
potentiation; HIV, human immunodeficiency virus; AMPA, a-amino-5-hydroxy-3-methyl-4-isoxazole propionic acid
* Corresponding author. Tel.: +34 91 585 47 43; fax: +34 91 585 47 53.
E-mail addresses: vgmarin@cajal.csic.es (V. Garcı́a-Marı́n), mfreire@cajal.csic.es (M. Freire).

0301-0082/$ – see front matter # 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.pneurobio.2007.06.002
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
1. Introduction
Santiago Ramón y Cajal (1852–1934) was one of the most
outstanding neuroscientists of all time. In 1906 he shared the
Nobel Prize for Physiology or Medicine with Camillo Golgi
(1834–1926) ‘‘in recognition of their work on the structure of
the nervous system’’. We think that the recent centenary of this
Nobel Prize offers a great opportunity to analyze one of Cajal’s
most important discoveries, the dendritic spines.
As proof of his enormous amount of work, 4529 histological
preparations that were personally made by Cajal are preserved
in the Museum Cajal. Of these preparations, 809 are stained
with the Golgi method and 108 with the Ehrlich method; these
methods allow seeing dendritic spines with the light micro-
scope. Apart from his preparations, more than 2500 original
drawings and papers written by Cajal – most of them in Spanish
or French – are conserved almost exclusively in the Cajal
Institute.1 In these funds there is a lot of information for
reinterpreting the discovery of the dendritic spines from a
historical and present-day point of view.
Dendritic spines are one of the most active fields of research
in modern neuroscience, but it took great effort before they
were considered as real structures. This review will deal in
depth with the scientific observations, reasoning, and ideas of
Cajal about dendritic spines, the historical context of the
discovery, and the meaning of Cajal‘s concept of dendritic
spines in current neuroscience.
2. The historical context of the discovery
‘‘. . . the surface . . . appears bristling with points or short
spines . . .’’2 (Cajal, 1888). This quotation, in Spanish, refers to
the dendritic spines of a Purkinje cell (Fig. 1), and marks the
beginning of the research into dendritic spines in the history of
neuroscience. In a footnote Cajal explained:
‘‘At first we believed that these protuberances were the result
of a tumultuous silver precipitation; but the constancy of
their existence . . . inclines us to consider them as normal
structures’’3 (Cajal, 1888).
The Golgi staining (Golgi, 1873) was the method used by
Cajal for visualizing and describing the fine structure of the
nervous system. Cajal was introduced to the Golgi method in
1887 by Luis Simarro La Cabra, the founder of clinical
psychology in Spain. The deep impression caused by this
staining made Cajal focus on the research of the nervous
system. One of the most important reasons for Cajal’s success
in these studies was that he applied the ontogenic method to his
1 (1) Constancy of the experimental results: Spines are always
Many of the works of Cajal have been tranlated to the English by DeFelipe
and Jones, in their books: Cajal on the Cerebral Cortex (1988) and Cajal’s
Degeneration and Regeneration of the Nervous System (1991).
2
‘‘. . . La superficie . . . aparece erizada de puntas o espinas cortas . . .’’.
3 4
‘‘Al principio creı́amos que estas eminencias eran resultado de una pre-
cipitación tumultuosa de la plata; pero la constancia de su existencia . . . nos
indica a estimarlas como disposición normal’’.
111
research reducing the complexity of the brain by applying the
Golgi staining to embryos and young animals.
But why were the dendritic spines discovered 15 years after
the development of the Golgi method? As we said before, the
Golgi method was developed by Golgi in 1873. Nevertheless,
this method had little repercussion in the scientific community,
probably because of its limited diffusion, the irreproducible and
apparently random impregnation, and something Cajal called
‘‘scholastic discipline’’:
‘‘out of respect for their teachers, students tend to use only
those research methods that have been developed by their
teachers themselves. As far as the great investigators are
concerned, they would feel dishonored working with other
people’s methods’’4 (Cajal, 1899a).
It was Cajal who noticed the value of this technique and
improved it. But the real genius of Cajal was to look with
different eyes than the rest of researchers, and to interpret the
histological observations in the correct manner. In the Golgi
preparations made by Golgi himself it is also possible to
observe dendritic spines (De Felipe, oral communication), but
he made no reference to them until the Nobel lecture in which
he mentioned the dendritic spines without ascribing them any
physiological significance. In addition, we have found only
three drawings of dendritic spines in Golgi’s work (Fig. 1H).
We may conclude from these data that Golgi also saw these
structures but that he did not give any importance to them,
maybe because he thought at first, as other researchers did, that
they were silver artifacts or because he did not think that they
had any physiological meaning.
The discovery of dendritic spines caused some controversy
in the scientific community. Some scientists supported Cajal’s
discovery with new data like Retzius (1891), Schaffer (1892),
Edinger (1893), Berkley (1896), and Monti (1895a,b), while
others denied the reality of these structures like von Kölliker
(1896), Meyer (1896a,b, 1897), and Dogiel (1896). They
argued that what Cajal called ‘‘collateral spines’’ were in reality
a silver precipitate, adhering to the widespread notion that the
Golgi method was not very reliable. It is interesting to note that
this idea was at first also considered by Cajal, but that he
quickly discarded it.
3. Principles of Cajal’s scientific reasoning
The discovery of the dendritic spines by Cajal provides a good
opportunity to analyze the principles of his reasoning in the
discovery of these structures, and his work trying to convince the
other scientists of the reality of the so-called collateral spines.
The following key points summarize the main ideas in
Cajal’s scientific reasoning:
absent from the soma and the origin of thick dendrites, and
‘‘por respeto al maestro, ningún discı́pulo suele emplear métodos de
investigación que no se deban a aquél. En cuanto a los grandes investigadores,
creerı́anse deshonrados trabajando con métodos ajenos’’.
112 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

Fig. 1. Dendritic spines of Purkinje cells: (A) first drawing of Ramón y Cajal showing the dendritic spines of a Purkinje cell of the hen (Cajal, 1888); (B) insert-box
showing the dendritic spines in real size; (C) Purkinje cell of an adult bird (P84186); (D) Purkinje cell of an adult human (P81750); (E) different types of dendritic
spines found on human Purkinje cells of human. From top to bottom: thin, mushroom, sessile, and ramified; (F, G) different segments of dendritic branches of Purkinje
cells of adult bird and human, respectively, showing different densities of dendritic spines; (H) drawing of a Purkinje cell by Golgi (1906), showing dendritic spines.
Golgi proposed that the dendritic tree directs its branches to the blood vessels, according to the nourishing role of the dendrites.

their distribution is not homogeneous along the dendritic
tree. If it had been a non-specific silver precipitate, it should
have been present on every part of the neuron with the same
intensity.
(2) Presence of dendritic spines in many species and many cell
types: Spines are observed on pyramidal cells of the
cerebral cortex, Purkinje cells, hippocampal pyramidal
neurons, dentate gyrus granule cells, etc., and in different
species (cat, dog, chicken, pigeon, human, etc.). After his
discovery in chicken Purkinje cells in 1888, Cajal extended
the observation to mammals: 15-day-old cat Purkinje cells
(Cajal, 1889) and rat olfactory granule cells (Cajal, 1890a).
One month later, in November of that same year, he
described dendritic spines in the cerebral cortex of lower
mammals. The first scientific drawing of human dendritic
spines appeared in ‘‘Nuevo concepto de la histologı́a de los
centros nerviosos’’ (Cajal, 1892). Cajal concluded that
dendritic spines are a common structure in many species
and many different types of cells, and that they might play
an important role in the functioning of the nervous system.
‘‘it is important to recognize certain morphologic details
on studying the protoplasmic expansions with the Golgi
method, because it is possible that in time they will be
understood to have physiological importance’’5 (Cajal,
1899b).
(3) Obtaining the same results using different staining methods:
Apart from the Golgi method, Cajal used the Golgi–Cox
method, Turnbull Blue method (Cajal, 1890b), and
Methylene Blue method (Cajal, 1896b) to visualize the
collateral spines of Purkinje cells. In the Golgi–Cox method
(Cox, 1891), silver nitrate is replaced by mercury chloride
and the tissue is next exposed to ammonia to darken the
resulting mercury precipitate (Zhang et al., 2003).
Visualization with Methylene Blue was considered the
definitive demonstration (Cajal, 1896b).
(4) Improvement and variations of the staining methods:
Because they were not able to stain them with Methylene
Blue, some researchers like Dogiel and Meyer denied the
reality of dendritic spines, arguing as done originally by von
Kölliker that they were silver artifacts. In fact, Dogiel
stained only just the beginning of the primary Purkinje cell
branches, where no dendritic spines are located, as can be
seen in his lithographic plate (Fig. 2A). Although Meyer
was able to stain the entire dendritic arbor, the color of the
drawing was pale blue (Fig. 2B), which led Cajal to think
that the staining obtained by Meyer was not strong enough.
Cajal had no doubt about the possibility of Methylene Blue
to stain the dendritic spines, because he had observed them
before in ganglion cells of the retina of the frog (Cajal,
1896a). This led Cajal to improve the method of Ehrlich,
and he published a monograph about dendritic spines
stained by Methylene Blue (1896b) (Fig. 2D and E) and
another important and more extended article later that same
year (Cajal, 1896b), in which he applied this staining to
many nerve centers (Fig. 5). The different results achieved
by these three scientists may be due to the different
Methylene Blue staining techniques they employed.
- Semi Meyer employed subcutaneous injection of Methy-
lene Blue into the living animal. Cajal tried to stain the
cortex and the cerebellum with this method, but the color
of the cells was too pale, and only the soma could be
distinguished. In Fig. 2C, a cell is shown from an original
preparation of Cajal impregnated with Methylene Blue,
done with the Meyer method and lubricated afterwards.
On the preparation Cajal wrote: ‘‘Rabbit . . . injection . . .
lubrify . . . 1 h . . . ganglia . . . ’’.6 No dendritic spines can
be seen because the staining is too pale. This cell is
comparable to an original drawing of Meyer (Fig. 2B).

5
‘‘Conviene conocer . . . porque acaso andando el tiempo alcancen tras-
6
cendencia fisiológica’’. ‘‘Conejo . . . inyeccion . . . lubrifica . . . 1hora . . . ganglios’’.
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130 113

Fig. 2. Different drawings and photos from preparations stained with Methylene Blue: (A) drawing of Dogiel (1896) of the cerebellum, showing the Purkinje cells
with only the soma and the primary branches stained; (B) drawing of Meyer (1896a) of a pyramidal cell without dendritic spines; (C) Z-projection (five sections) of a
pyramidal cell of a rabbit preparation by Cajal (P81392), stained with the Meyer method followed by lubrication of the tissue; (D) published drawing of Cajal (1896b)
showing pyramidal cells of cortex of adult rabbit stained with Methylene Blue; (E) Z-projection (10 sections) of a fragment of a dendrite with dendritic spines of a
granule cell of the dentate gyrus of rabbit (P81469), made using the Meyer method. On the label of the preparation is written (‘‘1 hora solo de aireación’’: only 1 h of
aeration); (F) different types of dendritic spines and filopodia seen in drawing (D). From top to bottom: sessile, thin, mushroom, branched, spine with spinule,
dendritic filopodia, and dendritic growth cone filopodia. The image is 1.25 times the real size of the drawing. On the right the same spine shapes found in a Golgi
preparation. (G) Dendritic branch (1.25 times real size) of the drawing compared with a dendritic branch in a Golgi preparation.

- Dogiel employed the technique of lubrication, the
Ehrlich–Dogiel method. After leaving pieces of nervous
tissue to the action of air, he applied a diluted solution of
Methylene Blue (Fig. 2A).
- Cajal used the propagation method created by himself.
The pieces were not exposed to the action of air, and the
color, as a pigment or in saturated solution, was applied
directly to the surface of the nerve tissue.
In the Museum Cajal there are 108 histological preparations
impregnated with Methylene Blue. In these preparations it is
very difficult to see the dendritic spines, probably because the
staining gets paler with time and because of varicose
degeneration. However, there are some preparations where it
is possible to find some dendrites with dendritic spines still
stained (Fig. 2E).
In contrast to von Kölliker, Dogiel, or Meyer there were
other scientists who did not deny the existence of these
structures, but who made a wrong interpretation of them. For
instance, Bethe did recognize their existence, but mistakenly
considered them as insertion points of an enigmatic interstitial
network of grey matter called nervöses Grau by Nissl. Held
also accepted them, but wrongfully assumed that they were the
endings of pericellular nerve fibers, calling them Endfüse
(Endfeet). But, as Cajal said, spines are never stained with
neurofibrillar methods like reduced silver nitrate (Cajal, 1909).
(5) Positive science: Negative results cannot invalidate positive
results. Cajal wrote:
‘‘Moreover, in good scientific logic, negative facts of
observation do not invalidate the positive ones. When a
method does not permit us to confirm a fact that can be
easily demonstrated with other methods, the only thing
we can state is that the method employed is not adequate
for the verification, as long as nobody proves in an
irrefutable way (and this has not been proven until now)
that the procedures for revealing the morphological
detail looked for cause artificial alterations in the cell’’7
(Cajal, 1896b).
7
‘‘Además en buena lógica cientı́fica, los hechos negativos de observación
no invalidan las observaciones positivas. Cuando un método no nos permite
confirmar un hecho, fácilmente demostrable con otros métodos, lo único que
legı́timamente podemos afirmar es la insuficiencia del recurso empleado para
hacer la verificación, á menos que no se pruebe de manera irrecusable (y esto
nadie lo ha probado hasta hoy) que los procedimientos reveladores del detalle
morfológico buscado, provocan en la célula alteraciones artificiales’’.
114 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

(6) Use of the best technical instruments: Cajal had a

microscope with the best optics of the time. He used very
powerful objectives. Using the Zeiss E 1.30 apochromatic
objective, he realized that the spines did not have the
appearance of crystals or irregular deposits. He also used a
Zeiss camera lucida, as he wrote in the footnotes of some of
his lithographic plates. The Camera Lucida gave Cajal the
opportunity to draw in a very realistic way, although he took
some artistic freedom by grouping cells of different
histological sections. However, contemporaries of Ramón
y Cajal who saw him drawing said that he used to draw by
looking at the histological preparation through the
microscope, while drawing on a piece of paper on his
right-hand side, where the exact reproduction of the nerve
cell appeared, without using the camera lucida that Cajal
considered troublesome (De la Villa, 1952). One possible
explanation of this way of drawing is that Ramón y Cajal
managed to blend the microscope image with the drawing
he was making on the paper seen with his right eye, so that
he did not need the camera lucida that specifically carries
out the function of mixing both images together (Freire, Fig. 3. Varicose degeneration: (A) original drawing of Cajal (1896a) of the
2003). fascia dentata of a 1-month-old rabbit, based on a preparation stained with
(7) The concept of scientific drawing in the work of Cajal: Methylene Blue. (B) Z-projection (five sections) of a granule cell of the dentate
gyrus of a rabbit with varicosities on its dendrites (P84169); (C) Z-projection
‘‘Good drawings, like good microscopic preparations, (11 sections) of dendritic branches with varicose degeneration.
are pieces of reality, scientific documents that preserve
their value indefinitely, and their reexamination will
always be profitable no matter the interpretations they
of prejudices and other people’s images, and to have the firm
may have elicited’’.8 (Cajal, 1899b).
intention to see and judge for ourselves’’9 (Cajal, 1899a).
In the act of drawing it is very important to maintain this
With this sentence he expressed very well his concept of
independent and open-minded way of seeing.
scientific drawing. Cajal tried not to act as a filter between what
he observed on his histological preparation and what the
4. Principal data contributed by Cajal to the research
spectator could observe on his scientific drawings. He did not
of the dendritic spines
try to interpret the structures he was watching, but to draw all
structures as realistically as possible, although there are other
The references and data contributed by Cajal about spines
schematic drawings where he tried to express a concept or give
are scattered throughout his different articles and books.
an explanation of what he was observing. One proof of this way
Moreover, the drawings related to dendritic spines help us to
of thinking is the drawings of the dendritic spines compared to
interpret the thinking of Cajal about these structures.
the drawings of the dendritic varicosities in the monoliforme
stage (Fig. 3). Cajal drew the varicosities and dendritic spines,
4.1. Morphological data
although he thought that the former were a post-mortem
phenomenon. In contrast to the scientific drawings of Cajal,
Cajal noticed the different morphologies of dendritic spines.
there are others from contemporary scientists that are more
This is in contrast to other scientists, who drew the dendritic
interpretations by the scientists than a representation of reality.
spines in a very cloned and symmetrical way like Golgi in his
There is another sentence that could help to understand this
Nobel lecture (mere sticks without a head). Instead, it is
idea.
possible to observe in the drawings of Cajal the three mayor
‘‘. . . to see things for first time, in other words, to types of spines described by Peters and Kaiserman-Abramof
contemplate them, putting aside what we learned from (1969): sessile, mushroom, and thin (Fig. 2F), and other
books, false descriptions and common platitudes, has great important features like ramified spines or spinules, structures
importance in scientific research. We have to free our minds

8
‘‘El buen dibujo, como la buena preparación microscópica, son pedazos de
realidad, documentos cientı́ficos que conservan indefinidamente su valor y cuya
revision será siempre provechosa, cualesquiera que sean las interpretaciones á
que hayan dado origen.’’
9
‘‘. . . ver las cosas por primera vez, es decir, readmirarlas, descartando
reminiscencias librescas, descripciones postizas y frases y tópicos comunes,
tienen en la investigación cientı́fica muy señalada aplicación. Hay que limpiar
la mente de prejuicios y de imágenes ajenas, hacer el firme propósito de ver y
juzgar por nosotros mismos . . .’’.
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
Fig. 4. Dendritic spines in the hippocampus: (A) original drawing of Cajal (1933) showing the mossy fibers that connect the granule cells of the fascia dentata with the
thorny excrescences of CA3; (B) three pyramidal cells of rabbit CA3 (P83474); (C) thorny excrescences of the apical shaft of a pyramidal cell; (D) different segments
(distal: up, with dendritic spines; and proximal: down, without dendritic spines) of a granule cell of a 6 days old cat hippocampus (P80714).
that may be implied in transendocytosis (Spacek and Harris,
2004). The morphology of the spine has great physiological and
functional importance, as we will see in the last section.
Cajal was the first to describe the thorny excrescences
(Cajal, 1893) (Fig. 4). These are clusters of dendritic spines
from CA3 and hilar cells. They are situated along the soma and
the apical dendrites of these pyramidal cells, although they can
be also present on the basal dendrites. They are the postsynaptic
structures of the mossy fibers of the granule cells.
Cajal also was the first to notice the differences in size of
dendritic spines among different areas like cortex and
cerebellum (Cajal, 1896a) (Fig. 5C).
‘‘In A, we show the appendages of a protoplasmic process of
a pyramidal cell of the mouse. It is striking to see the
slenderness and length of the pedicles that support the end
bulbs, and the many directions of the pedicles that sprout out
of the entire cylindric surface of the protoplamic branch. In
B we show the spined branches of the Purkinje cell of the
mouse. Note that the appendages are short and relatively
thick, and that they are close together and regularly spaced,
giving the process a hairy appareance’’.10 (Cajal, 1896a).
‘‘Their wealth, length and width vary among different cell
types: while they are thin and long in pyramidal cells, in
Purkinje cells they are short, thick and numerous’’.11 (Cajal,
1899b).
The differences in size of dendritic spines are not limited to
the cerebellum and the cerebral cortex. There are also
10
‘‘En A, figuramos los apéndices de una rama protoplásmica de una
pirámide cerebral del ratón. Llama la atención lo fino y largo de los pedı́culos
que sostienen los granos terminales y la diversa dirección de los mismos, los
cuales brotan de toda la superficie cilı́ndrica de la rama protoplásmica. En B,
mostramos las ramas espinosas de las células de Purkinje del ratón. Obsérvese
la cortedad y espesor relativamente considerable de los apéndices y su
proximidad y regularidad, que prestan a la expansión de que nacen un aspecto
festoneado’’.
11
‘‘Su riqueza, longitud y espesor, varı́an en los diversos tipos celulares; ası́
mientras en las células cerebrales, dichas espinas son finas y largas, en los
elementos de Purkinje, se muestran cortas, espesas y numerosas’’.
115
differences in size between distinct cortical areas suggesting
that the current injected by dendritic spines into the dendrite
changes depending on the cortical region and may have great
relevance in the functional specialization of the different areas
(Benavides-Piccione et al., 2002; Ballesteros-Yáñez et al.,
2006)
Cajal also made an observation of great value when he
compared the size of dendritic spines among species (Fig. 1E
and G). Comparing the dendritic spines of human and mouse,
he observed:
‘‘. . . the spines vary somewhat among different species; in
humans they are much longer and have thinner pedicles’’12
(Cajal, 1896a).
Recent studies have confirmed that the the head and the neck
in dendritic spines of human pyramidal cells are bigger and
longer respectively than the dendritic spines in the mouse
(Benavides-Piccione et al., 2002; Ballesteros-Yáñez et al.,
2006).
These data have great importance in modern neuroscience,
and form the basis of a great number of studies aimed at
establishing the differences of information processing between
different cortical areas and different species.
4.2. Distribution of spines
Spines are absent from soma and the origin of thick dendrites
(Fig. 4D).
‘‘The small terminal dendritic branches of the pyramids do
not have smooth contours, as the authors appear to represent
them; they bristle with teeth protruding more or less at right
angles (Figs. 5 and 7) and end in a round and slightly
thickened tip. These collateral spines are also found,
although in smaller numbers, on the large ascending shafts
of the medium and large pyramids, from the moment they
12
‘‘Por lo demás las espinas susodichas varı́an algo en las diversas especies;
ası́, en el hombre son mucho más largas y exhiben un pedı́culo más delgado’’.
116 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

Fig. 5. Different figures published in the article: ‘‘El azul de Metileno en los centros nerviosos’’ (Methylene Blue in nerve centers; Cajal, 1896a). (A) Fragment of the
Purkinje arborization of the mouse. According to Cajal: (a) opening for a blood vessel; (b) opening for climbing fibers. (B) Details of Purkinje dendritic spines. (C)
According to Cajal: varieties of spines of the cells of the molecular layer of the mouse cerebellum, stained with the Golgi Method: (A) spines of a pyramidal cell; (B)
spines of a Purkinje cell; (C) spines of a nest cell; (D) spines of a Golgi cell; (E) excrescences of a neuroglic cell of the molecular layer of the cerebellum; (a) large
openings for the astrocytes; (b) small openings for the parallel fibers.

begin emitting small branches in the deep layers of the
cortex’’.13 (Cajal, 1891).
This observation was a proof to Cajal of the reality of these
structures; if they were a non-specific silver precipitate, they
should appear randomly distributed over the entire neuron.
This absence of dendritic spines from the proximal region
of the dendrite has a great physiological importance. On
13
‘‘Les ramilles protoplasmiques terminales des pyramides nes sont pas lisses
de contour, comme les auteurs semblent les représenter; elles sont herissées de
dents naissant à angle droit, ou presque droit, Fig. 7 et 5, et terminées par un
bout rond et un peu éppaissi. On trouve aussi des épines collatérales, quoique
moins nombreuses, dans les grosses tiges ascendantes des moyennes et grandes
pyramides, depuis le moment oú elles commencent à émettre des ramilles dans
les couches profun, depuis le moment où elles commencent à émettre des
ramilles dans les couches profondes de l écorce’’.
these proximal regions only inhibitory synapses have been
found (Alonso-Nanclares et al., 2004). The distribution of
dendritic spines has been recently studied in detail by Elston
and DeFelipe (2002) in different cortical areas and species.
They found that: ‘‘. . . normalized cumulative spine distribu-
tion (as a function of relative distance from the soma to the
distal tips of the dendrites) is remarkably constant for all
cells’’ and ‘‘remarkably constant across species of different
orders’’. These authors propose two possible explanations for
the absence of dendritic spines at the origin of thick
dendrites:
- The mature membrane is nonpermissive to localization of
excitatory inputs.
- Characteristics of the backpropagation potential preclude
excitatory synapse localization on the soma and proximal
dendrites.
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
Another important observation of Cajal was the increased
density of dendritic spines in areas that are more innervated
(Fig. 5C).
‘‘A priori it is already conceived that, just like the form and
the number of nerve terminal fibers in each center of gray
matter vary, the disposition of the spined appendices
destined to make contacts will also vary in a correlative
way. To be certain that these accomodations are real, we
have made a comparative study of the spined appendices of a
great number of nerve- and neuroglic types’’.14 (Cajal,
1896a).
In this paper, he also made a comparison between different
types of cells and the connections made on these cell types
(Pyramidal cells, Purkinje cells, Nest cells, Golgi cells and
Neuroglic cells). Looking at the density of the spines, Cajal
reaches the following conclusion:
‘‘Such differences seem to indicate that the number of fibers
connected to Purkinje cells is much higher than the number
of fibers connected to nest cells and Golgi cells’’.15 (Cajal,
1896a).
This direct correlation between the number of spines and
the number of fibers is used in present-day neuroscience for the
comparative analysis of pyramidal cells and the analysis of the
complexity of cortical circuits and their evolution. Recent data
show that the cerebral cortex is characterized by regional and
species variations that support the different functions of distinct
cortical areas and the different cognitive capacity across
species. For instance the morphology of the pyramidal cell is
very complex and varies across different areas of the cerebral
cortex. The density of dendritic spines of pyramidal cells of the
prefrontal cortex (PFC) of humans and macaques is higher than
in sensory areas, for instance visual areas (Elston, 2000; Elston
et al., 2001; Elston and DeFelipe, 2002; Elston, 2003). The
increase in the complexity and the number of dendritic spines in
the pyramidal cells of the prefrontal cortex indicates a higher
capacity for the PFC cells to process and integrate information
of different sources that must be essential for the sustained tonic
activity characteristic of these neurons and their role in memory
and cognition. These studies still have a Cajal influence and
recall their statements about the pyramidal cell or psychic cell
(Cajal, 1892, 1894).
Other important data contributed by Cajal are about the
density of dendritic spines across species:
‘‘They vary also with animal species, and we may state
in general terms that a cell with spiny processes in
14
‘‘A priori se concibe ya que, variando como efectivamente varı́an en cada
foco de substancia gris la forma y el número de las fibrillas nerviosas
terminales, variarán también, de manera correlativa, la disposición de los
apéndices espinosos destinados a los contactos. Para asegurarnos de la
realidad de estas acomodaciones, hemos estudiado comparativamente los
apéndices espinosos de un gran número de especies nerviosas y neuróglicas’’.
15
‘‘Semejantes diferencias parecen indicar, si, como es muy probable, que el
numero de éstas relacionado con las células de Purkinje, es muy superior al de
las fibrillas conexionadas con los corpúsculos de cesta y los de Golgi’’.
117
homologous nuclei has more spines, the higher the level of
the subject in the animal serie. Thus, as an example, in
vertebrates the Purkinje cell of birds shows an arborization
less spinous than that of mammals’’.16 (Cajal, 1909)
(Fig. 1F and G).
The variation of the number of spines with the phylogenetic
scale is an issue of research in recent neuroscience. These
studies are important in order to establish how the cortical
microcircuity of the brain across species is conserved, and
which is the anatomical substrate that supports the different
cognitive and mental properties across species. Because the
number of spines represents the number of excitatory inputs
sampled by individual neurons, differences in spine density
represent also differences in patterns of neuronal connectivity:
the more spinous a neuron is the more capacity to process and
integrate information of different inputs. Elston et al. have
confirmed that the density of dendritic spines was consistently
higher in human compared with macaques, and higher in
macaques compared with marmosets for different areas of the
cerebral cortex indicating that the density of dendritic spines is
correlated with the brain size (Elston et al., 2001). Benavides
et al. have confirmed that the spine density of the human
temporal cortical neurons have on average a higher (30%)
spine density in the basal dendrite with the highest density of
spines than mouse temporal or occipital neurons (Benavides-
Piccione et al., 2002). However the pyramidal cells of the V1-
V2 area of the agouti (a large brain South American rodent
Drasypocta primnolopha) are 3–4-fold more spinous than those
of galagos, monkeys and baboons (Elston et al., 2006). These
differences in density across species and orders might be a clue
of the evolution of the brain in mammals. At least in a same
order the density appears to be higher the larger the brain is.
Across species of different orders the density of dendritic spines
in homologues areas may not be correlated with the size of the
brain.
In the case of the Purkinje cell filogenetic studies about
density in different species have not been done yet. Preliminary
studies based on the observation of the histological preparations
of Cajal of adult bird and human show that the human Purkinje
cells are more spinous than the bird ones as Cajal said (Fig. 1F
and G). However more quantitative data about spine density
within areas across species are essential for studying the
evolution and adaptation of the brain across species in different
orders.
4.3. Development
One of the most important methodological reasons for the
success of Cajal in his study of the nervous system was the
16
‘‘Elles varient meme avec ĺespèce animale, et nous pouvons, d’e une façon
générale, dire qu’une cellule à prolongements épineux est, pour des foyers gris
homologues, d’autant puls fournie d’épines, qu’elle appartient à un individu
puls´elevé dans la sèrie. Ainsi, pour ne prendre qúun exemple chez les vertebras,
la cellule de Purkinje des oiseaus a une ramure moins hérissée que celle des
mammifères’’.
118 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
Fig. 6. Dendritic spines and filopodia: (A) original drawing of Cajal (1933) showing filopodia and dendritic spines. According to Cajal: (A) shaft of a pyramid from
the visual region of a nearly mature rabbit; (B) pyramid of the visual region (2-month-old child); (C) shaft of a pyramid of a 1-month-old cat; (D) dendrite of a motor
cell of the spinal cord (1-month-old cat). (B–D) Photomicrographs of dendritic filopodia of the visual cortex of the cat (A, newborn; B, 9 days old; C, 12–15 days old;
E, 1-month-old).
application of the ontogenic method. Instead of studying the
complexity of the adult brain, he decided to study the nervous
system of lower animals and embryos, or of young higher
animals. Many of the Golgi impregnated preparations of the
Cajal Museum present filopodia instead of dendritic spines
(Fig. 6B–D). Dendritic filopodia are dendritic appendages that
may be implicated in spinogenesis, synaptogenesis, and
branching. They are usually present during development or
in pathological states.
This influence of age on the size of the dendritic appendages
was also noticed by Cajal:
‘‘Almost all their branches bear thin spines perpendicularly
inserted into their surfaces. These spines are stained in a
light coffee colour, and they are bigger than the terminal
branches of adult cells’’.17 (Cajal, 1889).
In this sentence, the principle of the shortening of the neck of
the spine during development can be recognized, which may
have functional consequences for the compartmentalization of
calcium by the spine.
There is another drawing that appeared in his book
Neuronismo ó Reticularismo? (Cajal, 1933) (Neuronism or
Reticularism?) (Fig. 6A), in which it is possible to recognize
spines with long necks that could be filopodia. In the text, Cajal
pointed out the difference in size of the dendritic appendages
depending on age:
‘‘I present in Figure 50 several drawings taken from the
shafts of young or adult cerebral pyramids. A represents the
shaft of a pyramid from the visual region of a nearly mature
rabbit. Note how short the spines are and how they begin
very thin and end in a terminal bulb. Very few of them are
bifurcated. In C the shaft of a pyramid of a one-month-old
17
‘‘Casi todas sus ramas poseen ligeras espinas perpendicularmente insertas
en su contorno. Estas espinas aparecen teñidas en café claro, y son más grandes
que las de las ramitas terminales de los corpusculos adultos’’.
cat is shown. Here the arrangement described in A is
confirmed; the spines appear a little bit longer and are
frequently curved. In B (two-month-old child) I have drawn
another shaft of a pyramid of the visual region. Here not only
the greater length of the appendages, but also the frequency
with which they divide and change the direction of their
secondary fibrils strike the attention. As a means of
comparison I have included the dendrite of a motor cell
of the spinal cord (D, one-month-old cat). Observe that the
surface is bristling with irregular projections that very
seldomly end in bulbs. It is almost certain that this
arrangement is transitory. Finally, I have drawn several
cruciform or oblique nerve collaterals of the cerebrum. It is
impossible to see any fusion of the latter with the spines’’.18
(Cajal, 1933).
It is possible to observe filopodia in the histological
preparations of Cajal (Fig. 6A). In the case of dendrite D of the
drawing, the appendages are surely filopodia: most of them do
not have a bulbous head and are highly ramified. Cajal thought
that these structures were transient.
In addition to these typical drawings, there are other
drawings (Fig. 7) of the histogenesis of the cerebellum
18
‘‘. . . mostramos en la figura 50 algunos dibujos tomados de tallos de
pirámides cerebrales adultas o jóvenes. En A presentamos el tallo de una
pirámide de la región visual del conejo casi adulto. Nótense cuán cortas son las
espinas y cómo empiezan delgadas y acaban por un bulbo final. Son pocas las
bifurcadas. En C copiamos un tallo de las pirámides del gato de un mes.
Confı́rmase la disposición mostrada en A; las espinas aparecen un poco más
largas y con frecuencia incurvadas. En B hemos dibujado otro tallo del niño de
dos meses (pirámide de la región visual). Llama la atención, no sólo la mayor
longitud de los apéndices, sino su frecuencia con que se dividen y los cambios
de dirección de sus ramillas secundarias. Como término de comparación hemos
dibujado en D una dendrita de una célula motriz de la médula (gato de un mes).
Adviértase que la superficie está erizada de proyecciones irregulares y rara vez
acabadas mediante bulbos. Es casi seguro que esta disposición es transitoria.
En fin, dibujamos también algunas colaterales nerviosas cruciales u oblicuas
(cerebro), sin que sea dable apreciar su fusión con las espinas’’.
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
Fig. 7. Dendritic filopodia and development in the cerebellum: original
drawing of Cajal (1904) of Purkinje cells at a very embryonic stage from
a newborn puppy. Golgi method. (A) superficial granules; (B) molecular
layer; (C) deep granules; (D) white matter; (a) Purkinje cell; (b) axon
collaterals of this cell; (g) embryonic granule. The Purkinje cells and granule
cells present multiple branches that could evolve from filopodia. In addition
around the soma and near the axon of the Purkinje cells there are other
appendages that we have confirmed like filopodia on the Cajal’s original
slides.
where it is possible to see some thin appendages that
may be filopodia transforming into branches. About the
development of the granule cells of the cerebellum, Cajal
comments:
‘‘Fine dendrites emerge now from the soma, which becomes
more and more roundish. These dendrites are initially long,
numerous, and poorly branched; finally some of these
appendages reabsorb, others become more regular and
produce the minute and specific arborizations at the tips,
characteristic of a fully developed granule’’.19 (Cajal,
1899a,b).
4.4. Physiological role of the dendritic spines
Ramón y Cajal made some hypothesis about the physio-
logical role of the dendritic spines. The clearest role attributed
by Cajal to dendritic spines was to increase the connective
surface:
‘‘. . . that by virtue of the aforesaid spines, dendritic branches
increase their receptive surface and establish more intimate
19
‘‘del soma, cada vez más espeso y redondeado, brotan ahora finas den-
dritas, las cuales son primitivamente largas, numerosas y apenas ramificadas; y
en fin, ulteriormente, algunos de tales apéndices se absorben, otros se reg-
ularizan, produciéndose en los extremos la diminuta y especı́fica arborización
del grano perfecto’’.
119
contacts with the axonal terminal arborizations’’.20 (Cajal,
1899a,b).
However, three-dimensional electron microscopic recon-
structions made by Harris and Stevens (1988) contradict this
view. By graphically removing all spines from their computer
reconstructions of spiny dendrites, they estimated that only 29–
45% of the dendritic membrane area of Purkinje cells would
have been covered by synapses if all spines were deleted and the
associated synapses moved to the dendrites. Applying the same
technique to five reconstructed CA1 pyramidal cell dendrites,
they found that only about 5–9% of the remaining dendritic
surface area would have been covered by the synapses from the
spines. Thus, it does not appear that dendritic spines are required
for lack of dendritic membrane area (Koch and Zador, 1993).
On the other hand, the idea of Cajal that dendritic spines
increase the connective surface allowing connections to axons
that are far way from the dendrite is very interesting:
‘‘In my feelings, the dendritic spines have the principal
function of increasing the surface of connection of the
protoplasmic arborization, going out to contact the nerve
fibres that are distant and cannot contact directly with the
outline of the dendritic process’’.21 (Cajal, 1896).
The dendritic spines may permit an axon to synapse with
dendrites of different neurons without a zig-zag trajectory, and
allow it to course through the neuropil in a relatively straight
path (Peters and Kaiserman-Abramof, 1969). It has been
corroborated that longer spines are occasionally found when
target axons are farther away into adjacent axon bundles in the
reticular nucleus of the thalamus and in the gelatinous
substance of the spinal dorsal horn (Fiala et al., 2002).
In his first papers Cajal thought about dendritic spines as
structures for canalizing the nerve fibers, which would press
down on the open spaces between spines:
‘‘The gulfs between such collateral spines receive impres-
sion from innumerable small fibers of the superficial layer.
Exactly the same disposition is possessed by the terminal
peripheral arborization of the large pyramids’’.22 (Cajal,
1890c).
But there are other statements of Cajal extracted from
‘‘Nuevo concepto de la histologı́a de los centros nerviosos’’ that
prove that Cajal saw the contacts between the nerve fibres and
the dendritic spines and thought they might be implicated in
transmission:
20
‘‘Que por virtud de las susodichas espinas, la ramificación protoplásmica
aumenta su superficie colectora y se establecen contactos más ı́ntimos entre
aquélla y las arborizaciones nerviosas terminales’’.
21
‘‘En mi sentir, las espinas tienen por principal oficio aumentar las super-
ficies de conexión de la arborización protoplásmica, saliendo al encuentro de
las fibras nerviosas que, por hallarse á cierta distancia, no pueden establecer
contacto directo con el contorno de las prolongaciones dendrı́ticas’’.
22
‘‘Los golfos que median entre tales espinas colaterales reciben la impresión
de las innumerables fibrillas de la zona superficial. Exactamente igual dis-
posición posee la arborización periférica final de las grandes pirámides.’’
120 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

‘‘. . . The existence of collateral spines on the dendritic

process; and the connection of all [the spines],23 at the level
of the molecular layer, with a tight plexus of little terminal
nerve fibers’’24 (Cajal, 1892, 1894).
Later on in the Textura (Cajal, 1899a,b), Cajal accepted the
function of connection of dendritic spines proposed by Berkley,
because it reconciles well with his hypothesis exposed in other
works:
‘‘Do they represent the lines of charges or absorption of
nerve impulses as stated by Berkley? The latter opinion
appears plausible to us. It reconciles well with our idea
expressed in another publication, namely that by virtue of
the aforesaid spines, dendritic branches increase their
receptive surface and establish more intimate contacts with
the axon terminal arborizations’’.25 (Cajal, 1899a,b).
As Cajal said, Berkley clearly defines dendritic spines as
receptors of currents:
‘‘The function of the gemmule is in all likelihood to receive Fig. 8. Similar drawings of (A) Berkley (1896) and (B) Cajal (1933) showing
nerve impulses from the endings of the numerous terminal connections of axons with dendritic spines.
nerve fibers that seem almost to touch them, and carry these
impressions to the dendrite and by its medium on the cell
body. Differences in the function of the gemmule of the receptors and it was his merit to localize the reception of the
pyramidal and Purkinje cell are probable’’ (Berkley, 1895) nervous current on the bulbous head of the spines:
(Fig. 8A).
‘‘. . . these spherical apparatus (terminal buttons) are closely
In his final work Neuronismo o Reticularismo? Cajal did not adjusted against the bulbous tips of the gemmules, at times
clearly point out the possibility of dendritic spines as receptors the application being so close as to give the impression of
of nerve impulses, and he only stated that nerve fibers of the actual contact . . . the axonal discharges of stimuli overleap
cerebral cortex rest on the spines. the infinitesimal distance between bulb and gemmule . . .’’
(Berkley, 1897)
‘‘Note how these collaterals cross and enter into transversal
or oblique contact with a great number of the dendritic Cajal did not specify where these contacts were made (head
shafts. It is probable that collaterals rest on the spines wich or neck). Today it is known that the majority of excitatory
cover the protoplasmic surface like down’’.26 (Cajal, 1933) synapses are made on the head of the spine, although sometimes
(Fig. 8B). there are synapses on the neck of the head that usually are
inhibitory (Jones and Powell, 1969). In contrast to Berkley,
We should take into account the contribution of Berkley
Cajal’s statements suggest that the contacts could be made also
clarifying the role of dendritic spines as nervous current
on dendritic shafts. Today it is known that the majority of
excitatory connections in the cerebral cortex are made on the
dendritic spines (Peters and Kaiserman-Abramof, 1969, 1970),
23
‘‘These words (the spines) do not appear in the spanish edition ‘‘Nuevo although excitatory inputs can be made also on dendritic shafts
concepto . . .’’, but appear in the french version Les Nouvelles Idées sur la (Hersch and White, 1981; White and Hersch, 1981), especially
Structure du Système Nerveux chez l’Homme et chez les Vertébrés: ‘‘. . . on Layer VI pyramidal cells (Mc Guire et al., 1984). In addition
l’existence d’épines collatérales sur les branches protoplasmiques et la con-
nexion de toutes ces épines au niveau de la zona moléculaire, avec un plexus
Berkley thought that nervous conduction was only longitudinal
serré de fibrilles nerveuses terminales’’ (Cajal, 1894) (see also De Felipe and and occurred between the bulbous terminations of the nerve
Jones, 1988). fibers that he had described with the silver phosphomolibdate
24
‘‘La existencia de espinas colaterales en las ramas protoplásmicas, y la method and the bulbous heads of the gemmule of the pyramidal
conexión de todas ellas, al nivel de la zona molecular, con un plexo tupido de cells (Berkley, 1896). Cajal did not agree with the hypothesis of
fibrillas nerviosas terminales’’.
25
‘‘Representan lı́neas de carga ó de absorción de corrientes nerviosas, como dendritic spines as the only place for receiving nerve impulses
declara Berkley? Plausible nos parece esta última opinión, que por otra parte, because it could not explain the connections, for instance,
se concilia bien con la idea expuesta por nosotros en otro trabajo, á saber: que between the pericellular axon arborization and the soma or the
por virtud de las susodichas espinas, la ramificación protoplásmica aumenta su origin of axons of the Purkinje cells that lack dendritic spines,
superficie colectora y se establecen contactos más intimos entre aquella y las or between the climbing fibers and the primary and secondary
arborizaciones nerviosas terminales’’.
26
‘‘Nótese cómo dichas colaterales cruzan y entran en contacto transversal y branches of the Purkinje cell. Cajal thought that in these
oblicuo con gran número de tallos de dendritas, apoyándose quizás en las articulations there might be a horizontal transmission between
espinas que revisten, como un vello, las superficies protoplásmicas.’’ the nerve fibers and the soma (Cajal, 1899a,b). Afterwards,
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130 121

apart from these end-bulbous terminals described by Berkley,

en passant varicosities were also described (Colonnier, 1968).
Stefanowska, another contemporary scientist of Cajal,
proposed the idea that what Stefanowska called the pyriform
appendages could modify their form and dimension during life,
loosening their contacts with nerve fibers to a variable degree
and explaining the sleep state by the disappearance of the
pyriform appendages and the interruption of the transmission
(Stefanowska, 1897a,b). This hypothesis was very much
influenced by the theory of neuronal ameboidism of Duval
(Duval, 1895), based on Rabl-Rückhard’s previous studies
(Rabl-Rückhard, 1890). This hypothetical retraction of the
spines is the first proposal that the dendritic spines are motile
until the theoretical work of Crick (1982) and the final
definitive observation made by Fischer et al. (1998).
The function and physiological role of dendritic spines Fig. 9. (A) Original drawing of Cajal (1904) of a section of the olfactory
continues being a subject of research. The definitive bulb of a several days old kitten, (I, J, granule cells). (B) Olfactory bulb of 1-
month-old mouse (granule cell and mitral cell). The insert box does not
demonstration of dendritic spines as postsynaptic units came
correspond to this preparation. It corresponds to a granule cell of newborn
from the electron microscopic studies of Gray (1959a,b). Many dog.
other functions have been proposed because excitatory inputs
can be made on dendritic shafts without dendritic spines
(Hersch and White, 1981; White and Hersch, 1981). A great list
of functions have been attributed to these important structures: In explaining the current flow in these circuits, he explained:
synaptic plasticity, biochemical compartmentalization (Ca2+
‘‘The peripheral process would represent, if not morpho-
and other metabolites), neuroprotection, increasing of the
logically at least dynamically, a functional process since
dendritic membrane capacitance, intersynaptic isolation,
nerve impulses circulate through it in the cellulifugal sense,
attenuation of passive synaptic potential, linear summation
as they do in true axons’’.28
of EPSPs, generation of rapid local EPSPs (for a review see
Shepherd, 1996). Reciprocal synapses between granule cell spines and mitral
A special case of dendritic spines are those of the olfactory cell dendrites are implicated in mediating feedback and lateral
bulb granule cells where dendritic spines also behave as inhibition of the mitral and tufted cells (Rall et al., 1966; Rall
presynaptic elements. The dendritic spines of the olfactory and Shepherd, 1968). The lateral inhibition through the granule
granule cells establish reciprocal synapses with the dendrites of cell spines enhances the tuning specificity of odor responses
the mitral cells (Rall et al., 1966; Rall and Shepherd, 1968; contributing to discrimination of olfactory information (Yokoi
Prince and Powell, 1970; Shepherd and Brayton, 1979). These et al., 1995).
spines (gemmules) receive type I excitatory synapses from
mitral cells and have reciprocal type 2 inhibitory synapses onto 4.5. Dendritic spines in pathological and poisoning states
those same dendrites. They are considered as the smallest
neuronal compartment capable of performing a complete After the verification of dendritic spines as real structures,
input–output operation of a single synapse (Shepherd and many scientists investigated their morphology in different
Greer, 1988). It should come as no surprise that this also began pathological states. The most typical alteration of dendritic
with Cajal. Cajal described the dendritic spines of the olfactory spines is the monoliforme state or varicose degeneration. It
bulb granule cells in 1890 (Cajal, 1890a). Later on, in the consists in the formation of swellings along the dendrites and
Textura (1899) Cajal confirms the absence of axon in granule the absorption of the spines (Fig. 3). Surprisingly, Golgi was the
cells as was first described by Golgi (1875). Describing the first to describe varicose degeneration in a case of Chorea
dendrite of the granule cells Cajal wrote: (Golgi, 1877).
This alteration was described in general paralysis (Colella,
‘‘. . . the peripheral process has a constant orientation and
1892), melancholia (Azoulay and Klippel, 1894), epileptic
connection . . . it terminates in a tuft of very spiny branches
dementia (Colella, 1892), septicemia, haemorrhage (Tirelli,
in contact with the secondary dendrites of mitral cells.’’27
1895), alcoholism, and serum and ricin poisoning (Berkley,
(Cajal, 1899a,b) (Fig. 9).
1895) (Fig. 10C); exposure to chloroform, ether, poison gas,
and electrocution were also found to lead to these alterations

27
‘‘la expansión periférica de los granos posee una orientación y conexión
invariables. . ..se termina á favor de un penacho de ramas fuertemente espi-
nosas en contacto con las dendritas secundarias nacidas de las células
mitrales’’.
28
‘‘El apéndice periférico representarı́a, si no morfológica, dinámicamente,
una expansión funcional, puesto que la corriente nerviosa circula en él en
sentido celulı́fugo, como en los axones legı́timos’’.
122 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

Fig. 10. (A) Drawing of Monti (1895b) showing varicose degeneration of a pyramidal cell of the dog. The degenerated processes are those directed towards the blood
vessels. Axons never present this varicose degeneration. (B) Drawing of Stefanowska (1897a) showing varicose degeneration because of the action of ether. (C)
Drawing of Berkley (1895) of a Purkinje arborization of a control cell (left) and a cell subject to chronic alcohol poisoning (right). (D) Camera lucida drawings (A, B:
Marı́n-Padilla, 1972; C, D: Purpura, 1974) of apical dendrites of pyramidal cells from human cerebral cortex. (A) apical dendrite of a newborn girl with D1(13–15)
trysomy (Patau syndrome); (B) apical dendrite of an 18-month-old mentally retarded with 21 trysomy (Down syndrome); (C) a dendrite from normal 6-month-old
infant with no history of neurological disorder with a large number of normal spines; (D) analogous dendrite from a retarded 10-month-old child, showing long,
tortuous spines; (E) schematic of different spine abnormalities in different pathologies (Fiala et al., 2002).

(Stefanowska, 1897a) (Fig. 10B). Very interesting are the works
of Monti (Fig. 10A), describing varicose degeneration in
embolism and starvation (Monti, 1895a,b). He used these data
to support the hypothesis of Golgi about the nourishing function
of dendrites and the conductive role of the axons, opposing the
theories of Cajal.
For Cajal, varicose degeneration represented a post-
mortem alteration that occurs because the tissue pieces are
fixed too late. This disorganization of the dendrites is
eliminated if the tissue is fixed rapidly, but it is more difficult
to avoid in the Ehrlich staining because it needs the
participation of air in the staining process (Fig. 3). Another
important piece of data is that this degeneration preferen-
tially occurs in thick pieces of nervous tissue in the deep
layers, the areas that are stained later (Cajal, 1899b). Cajal
did not deny the possibility of these alterations under
pathological states, although he was very cautious about it
because he discovered dendritic spines in animals anesthe-
tized by chloroform.
The swelling and loss of spines is nowadays accepted to
occur in epilepsy, hypoxia/ischemia, traumatic injury, edema,
and acute excitotoxicity (for a review, see Fiala et al., 2002)
(Fig. 10E). In addition, loss of spines has also been described in
poisoning, alcohol abuse, and epilepsy. It was difficult to decide
if this varicose atrophy was real or a degenerative post-mortem
process, not related with the disease.
5. The dendritic spines from Cajal to present-day
neuroscience
5.1. The concept of Sherrington’s synapse
It was Sherrington who introduced a concept that would
change neuroscience for years to come, the concept of synapsis.
‘‘So far as our present knowledge goes, we are led to think
that the tip of a twig of the arborescence is not continuous
with but merely in contact with the substance of the dendrite
or cell body on which it impinges. Such a special connection
of one nerve cell with another might be called a synapse’’
(Foster and SherrinGton, 1897; see also Shepherd, 1997).
This proposal united neuroanatomical and physiological
evidence into a single term. Cajal accepted this term and he
used it in his last book Neuronismo o Reticularismo? (Cajal,
1933).
The definitive identification of the synapse was done using
electron microscopy (Palay, 1956). In 1959, Gray identified the
dendritic spines as postsynaptic structures. He also classified
synaptic contacts into type I (made on dendritic trunks and
dendritic spines) and type II (made on dendritic trunks and
soma). The type I synaptic contacts have a thicker postsynaptic
density. These types of synapses correspond to the asymmetric
and symmetric types of Colonnier (1968). The importance of
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
this classification resides in its functional implications:
asymmetric spines are excitatory and glutamatergic, while
symmetric synapses are inhibitory and gabaergic.
5.2. Morphology of dendritic spines
The observation of dendritic spines by electron microscopy
showed that they contain flocculent material and endoplasmic
reticulum (ER). When the ER cisternae are located in groups
longitudinally oriented away from each other, they form the so-
called spine apparatus first described by Gray (1959a,b); the
spine apparatus has many important functions like Ca2+
regulation. In addition some other elements may be present:
sparse Golgi elements (Horton et al., 2005), mitochondria
(Adams and Jones, 1982), endosomes, (Cooney et al., 2002)
proteosome (Ehlers, 2003), and filaments of actin (Fifkova and
Delay, 1982).
After Gray, ultrastructural studies were continued by Jones
and Powell (1969) and by Peters and Kaiserman-Abramof
(1969), who classified the different shapes of the dendritic
spines into thin (more abundant), sessile, and mushroom spines
(Fig. 2F). This classification is still useful for investigating the
structure-function relationships of dendritic spines, although it
has been hypothesized that there may be a continuum of shapes.
In addition, it is difficult to assign some spines to any of these
groups because they have intermediate shapes. Indeed, there
appears to be a clear relationship between the morphology and
the function of the spine, particularly with relation to the size of
the spine head and the length of the neck. For instance, the
volume of the head is directly proportional to the size of the
postsynaptic density (Freire, 1978), to the number of
postsynaptic receptors, to the size of the presynaptic terminal
(Špaček and Hartmann, 1983; Peters, 1987), to the presynaptic
number of docked synaptic vesicles, and to the readily
releasable pool of neurotransmitters (Špaček and Hartmann,
1983; Harris and Stevens, 1988; Nusser et al., 1998; Schikorski
and Stevens, 2001). Recent studies have shown that the number
of AMPA receptors in the postsynaptic density is directly
correlated with the size of the head (Matsuzaki et al., 2001;
Kasai et al., 2003), and that this influences the capacity of being
potentiated by long-term potentiation (LTP).
Important data regarding the structure of the spine and its
function have come from living image studies of the cerebral
cortex of mice (Trachtenberg et al., 2002; Grutzendler et al.,
2002; Matsuzaki et al., 2001, 2004; Kasai et al., 2003). These
studies have shown that big dendritic spines are more stable
than small ones, and that they can persist for months. These
data, combined with the data on AMPA receptor density and
LTP research, have led Kasai to propose the hypothesis of
learning and memory spines. Big spines are stable and
represent the physical trace of long-term memory by forming
stable synaptic connections (memory spines), while thin
spines are motile and instable and form weak connections
implied in learning. It has also been shown that the number of
memory spines or big stable spines is higher on the dendrites
of old animals than on those of young animals (Holtmaat
et al., 2005).
123
Another important issue is the length and width of the neck.
It has been postulated that the neck of the spine could reduce
the weight of the synapse (Chang, 1952): ‘‘If the end bulbs of
the gemmule (spines) are the receptive apparatus for the
presynaptic impulses, the process of postsynaptic excitation
initiated there must be greatly attenuated during its passage
through the items of the gemmules which probably offers
considerable ohmic resistance because of their extreme
slenderness’’. Especially interesting are the theoretical works
based on computational models made by Rall and Rinzell (Rall,
1964; Rall and Rinzel, 1971a,b; Rall, 1974). They introduced
the idea that the spine might be a site for neuronal plasticity.
‘‘. . . fine adjustments of the stem resistances of many spines . . .
could provide an organism with a way to adjust the relative
weights of the many synaptic inputs . . .; this could contribute to
plasticity and learning of a nervous system’’ (Rall and Rinzel,
1971b). Crick also thought that the contraction and extension of
the neck could change the weight of the synapses (Crick, 1982).
However, this role of the neck in the reduction of the weight
of the synapses was questioned by Koch and Zador (1993), who
postulated that the spine neck conductance appears to be too
large relative to the synaptic conductance change to provide
effective modulation of the amplitude of the synaptic current
generated at the spine head. Instead they proposed that spines
create an isolated biochemical microenvironment around the
synapse, so that the spines function as biochemical compart-
ments. Because of its narrowness, the neck can be considered a
barrier for the diffusion of metabolites isolating the dendritic
spines biochemically from the dendrite (Svoboda et al., 1996;
Majewska et al., 2000a; Nimchinsky et al., 2002; Segal, 2005).
One of the more important metabolites in glutamatergic neuronal
transmission is Ca2+ that can act as a second messenger in
different signaling pathways implicated in plasticity functions
such as memory and learning (Yuste and Denk, 1995; Segal,
1995; Korkotian and Segal, 1999; Majewska et al., 2000a,b;
Yuste et al., 1999; Yuste and Holthoff, 2000; reviewed in Yuste
and Bonhoeffer, 2001; Yuste and Bonhoeffer, 2004; Hayashi and
Majewska, 2005; Segal, 2005; Konur and Ghosh, 2005; Oertner
and Matus, 2005; Ethell and Pasquale, 2005). In addition, the
concentration of Ca2+ in the spines can be considered to be under
the control of different buffer systems (Calmodulin; spine
apparatus). It has been observed that spine apparatuses are
present preferentially in the base of the neck of mushroom spines
while they are absent from many of the thin spines, possibly
regulating in this manner the influx of calcium into the spines.
5.3. Dendritic spines and pathology
Data about the possible functions of dendritic spines have
also come from studies of pathological states of the brain. In
these pathological states, as was first proposed by contemporary
scientists of Cajal, there is a significant loss of dendritic spines.
Changes of dendritic spines in pathological states have been
described in many alterations such as malnutrition, brain
edema, traumatic lesions or tumors, alcoholism, drug con-
sumption, poisoning, epilepsy, transmissible diseases (HIV,
Creutzfeldt-Jakob, Kuru, etc.), dementias (Alzheimer’s, Pick’s,
124 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
Huntington’s and Parkinson’s diseases), and mental pathologies
(schizophrenia and major depression, etc.) (for a review, see
Fiala et al., 2002). We will focus on only two topics: mental
retardation and hypoxia.
Marı́n Padilla used the Golgi method to study the motor
cortex of a newborn girl with D1 (13–15) trisomy (Patau
syndrome) and the motor cortex of an 18-month-old girl with
21 trisomy (mongolism or Down syndrome) (Marı́n-Padilla,
1972) (Fig. 10D). He described loss and distortion of dendritic
spines in the Patau syndrome. Dendritic spines were irregular,
long, and tortuous and appeared disoriented with swellings
along their surfaces that could make immature synaptic
contacts. Thus, they are very similar to immature appendages.
He proposed two explanations for this immaturity: appendages
fail to convert to dendritic spines, or there are too few axons to
mature the appendages (Marı́n-Padilla, 1974). In Down
syndrome, Marı́n-Padilla (1976) found three different patterns
of spine pathology in pyramidal neurons of the motor cortex, in
addition to a few essentially normal neurons. Some neurons
were densely covered with long, tortuous spines. Other neurons
were uniformly covered with short, thin spines that were
abnormally small in volume. A third type of neuron exhibited
significant spine loss, with the few remaining spines having
very large heads and thin necks. A pattern of small spines in
neonates and abnormally long spines in older infants has been
observed in more extensive studies of Down’s syndrome
(Takashima et al., 1994). Purpura observed a reduction of the
number of spines and the presence of spines with very long
necks and big heads in children with a normal karyotype but
with deep mental deficiency of unknown etiology (Purpura,
1974) (Fig. 10D). Unusually long, tortuous spines occur in
many other forms of mental retardation, including Fragile X-
Syndrome (Rudelli et al., 1985; Segal et al., 2003), Maple
Syrup Urine Disease (Kamei et al., 1992), and fetal alcohol
syndrome (Ferrer et al., 1987).
The similarity of spine pathologies in different conditions
associated with mental retardation is striking, and led to the
recent suggestion that the different genetic deficits associated
with mental retardation disrupt a common signaling pathway
related to the development of the dendritic cytoskeleton
(Martone et al., 2000).
Hypoxia or ischemia leads to the formation of dendritic
varicosities, with a corresponding loss of spines. This damage is
principally mediated by the high release into the extracellular
space of glutamate, which permanently activates NMDA
receptors causing a massive entrance of Ca2+ into the
intracellular space. However, this phenomenon may be
reversible. Amazingly, spines absorbed in this manner can
recover their original shape after termination of the insult and
elimination of dendritic swelling, suggesting that the synapses
are not lost during this process; this may involve a mechanism
where dendritic spines are not formed from pre-existing
filopodia, but from pre-existing synapses contacted by axons
(Hasbani et al., 2001). However, ischemic lesions due to stroke
in the mature CNS can produce more permanent spine loss (De
Ruiter and Uylings, 1987). As we have seen before, Cajal noted
that it is possible to observe the swelling of dendrites and
resorption of spines during the first half-hour after exitus (Cajal,
1899a,b). Biopsy material, fixed quickly, is surely much better
preserved. In any case, the time interval between withdrawal
and fixation of post-mortem brain tissue is very important to
avoid artifactual swelling.
5.4. Plasticity of dendritic spines
Other interesting lines of research have come from
experiments on plasticity associated with changes in spine
density, initiated by Demoor (1896) and Stefanowska (1897a,b)
and recuperated later by Globus and Scheibel (1967): they
found a loss of dendritic spines in the apical shafts of the
neurons after eye enucleation and lateral geniculate lesions.
Valverde (1967, 1968, 1971) observed using the Golgi method
that light-deprived mice had a reduced number of spines in the
visual cortex that was higher in young animals than in older
animals, although the exponential increase of the dendritic
spines in normal animals was maintained in the light-deprived
animals. Moreover, these changes were completely reversible
in some neurons while other neurons remained damaged
(Valverde, 1971). A possible explanation came from the
realization that there are populations of spines with dimensions
too small to be seen with the light microscope, and that can be
seen only with electron microscopy (Freire, 1978). The
formation and maintenance of dendritic spines depends on
synaptic activity, and these changes may be modulated by
sensory experience (Valverde, 1967, 1968, 1971; Globus and
Scheibel, 1967; reviewed by Lippman and Dunaevsky, 2005;
Matus, 2005; Segal, 2005). Moreover, it has been shown that
early exposure to a rich foreground produces an increase in
dendritic arborization and the number of dendritic spines
(Rosenzweig et al., 1972; Volkmar and Greenough, 1972;
Globus et al., 1973; Greenough et al., 1973).
These conclusions clearly bring to mind Cajal’s concept of
plasticity and mental exercise:
‘‘The remarkable intellectual increase that is observed in
men dedicated to deep and continuous mental exercise; and
the coexistence of a remarkable talent or even of true genius
with medium-sized brains or brains of smaller than normal
dimensions and weights. In the first case, since the brain
cannot produce new cells (nervous cells do not multiply like
muscle cells do), it can be assumed that brain gymnastics
take the unfolding of the protoplasmatic expansions and the
nervous collaterals a little beyond what is common, forcing
the establishment of new and more extensive intercortical
connections [. . .].’’29 (Cajal, 1892).
29
‘‘El notable acrecentamiento intelectual que se observa en los hombres
consagrados á un ejercicio mental profundo y continuado; y la coexistencia de
un talento notable y aun del verdadero genio con cerebros de tamaño medio ó
inferiores á la dimensión y pesos normales. En el primer caso, podrı́a suponerse
que la gimnasia cerebral, ya que no puede producir células nuevas (las células
nerviosas no se multiplican como las musculares) lleva un poco más allá de lo
corriente el desenvolvimiento de las expansiones protoplasmáticas y colater-
ales nerviosas, forzando el establecimiento de nuevas y más extensas conex-
iones intercorticales [. . .].’’
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
The most important data about the plasticity of these
structures and its possible implication in functions as learning
and memory come from studies on LTP, inspired in the works of
Rall and Rinzel (Rall, 1974; Rall and Rinzel, 1971a,b). During
LTP there is an increase in the volume of the spine head (Van
Harrefeld and Fifkova, 1975; Fifkova and Van Harreveld, 1977)
and a variation of the neck length (Fifkova and Anderson,
1981). Engert and Bonhoeffer (1999) combining a local
superfusion technique with two photon imaging have shown
that new spines can appear on the postsynaptic dendrite after
LTP. In adition during LTP, the number of AMPA type
glutamate receptors at the spine head membrane increases due
to enhanced transport from recycling endosomes (Park et al.,
2004), acting as a mechanism for the potentiation of
transmission. In recent elegant experiments, Park et al.
(2006), using a combination of serial electron microscopy
and live cell fluorescence microscopy, have confirmed that
these recycling endosomes also provide membrane for activity
dependent spine growth and remodeling. Korkotian and Segal
(2007) have shown that chemical induction of LTP increases the
concentration of GluR1 in the head of the spines, preferently
those with short neck, which is F-actin and protein synthesis
dependent. In addition to the intracellular routes, glutamate
receptors can be transported to the spine head by lateral
diffusion along the membrane (Triller and Choquet, 2005;
Ashby et al., 2006). The molecular mechanisms that regulate
the vesicular trafficking to the spine membrane are unknown.
Endosomal transport and vesicular fusion are Ca2+ sensitive
(Khvotchev et al., 2003). CamKII another central intracellular
signal for LTP may regulate other effectors of endosomal traffic
(Karcher et al., 2001). Another signaling molecule for LTP,
cAMP dependent protein kinase (PKA) regulates endosomal
traffic and activity-induced insertion of AMPA receptors in the
spines membrane (Ehlers, 2000). Future research should focus
on the molecular mechanisms on endosomes and endosmal
traffic effectors that sense and respond to NMDA receptor-
mediated Ca2+ influx during LTP.
Another proof of the plasticity of dendritic spines is the fact
that they move, as first demonstrated by Fischer et al. (1998),
using cultures of neurons expressing GFP. Motility of spines
was first proposed by Demoor (1896) and Stefanowska
(1897a,b). In 1982, Crick hypothesized about possible move-
ments of the spines, and its implication for memory in an article
entitled: ‘‘Do spines really twitch?’’ He wondered: ‘‘Suppose
actin was discovered in dendritic spines?’’ This discovery was
in fact made 5 years earlier by Blomberg et al. (1977), who
identified actin in the postsynaptic density fraction isolated
from dog cerebral cortex. The definitive observation was made
by Fifkova and Delay (1982). Recent advances using two
photon microscopy have shown that the movement of spines
also ocurs in brain slices (Dunaevsky et al., 1999) and in intact
brains of anesthetized animals (Lendvai et al., 2000).
The actin cytoeskeleton is essential for the movement of the
spine, its shape and for synaptic plasticity (Matus et al., 2000).
Okamoto et al. (2004) have demonstrated that LTP induces
remodelation of spine actin cytoeskeleton. Tethanic stimulation
causes a rapid, persistent shift of actin equilibrium toward
125
filamentous actin (F-actin) in the dendritic spines of rat
hippocampus. This enlarges the spines and increases post-
synaptic binding capacity. In contrast, prolonged low frequency
stimulation shifts the equilibrium towards globular actin (G-
actin) resulting in a loss of postsynaptic actin and of structure
(Okamoto et al., 2004). These experiments indicate that F-actin
acts as a support for a larger spine. However, for the increase of
volume observed in LTP apart from the actin cytoeskeleton new
sources of membrane are required. This new membrane could
be provided by the recycling endosomes (Park et al., 2006).
Furthermore, the actin cytoeskeleton could be an ideal
mechanism for the traffic of the postsynaptic proteins and
lipids to their targets. In support of this hypothesis is the
abundant existence of actin-based myosin motors and myosin
regulatory proteins in the dendritic spines, (Osterweil et al.,
2005; Ryu et al., 2006).
The assembly-disassembly of actin filament is regulated by
actin-binding proteins (Arp2/3 complex, Cortactin, ADF/
Cofilin, Drebrin, Profilin II, Gelsolin, Spinophilin, etc.) (for
a review see Ethell and Pasquale, 2005; Tada and Sheng, 2006).
Many signaling pathways that regulate dendritic spine shape
and motility originate at the cell surface and converge on these
actin regulatory proteins like for instance the Rho/Rac
GTPases, Ras GTPases/MAPKinase, Ca2+ signaling (Ethell
and Pasquale, 2005; Tada and Sheng, 2006)
The process of molecular plasticity on dendritic spines is
very complex and dependent of many factors. Furthermore,
local protein syntheis can also contribute to the synaptic
plasticity. Poliribosomes redistribute from dendritic shafts into
spines during LTP (Ostroff et al., 2002; Bourne et al., 2007). In
contrast to the changes in the postsynaptic density, presynaptic
mechanisms (e.g. increase transmitter release) could mediate
the initial phase of potentiation in LTP.
5.5. Dendritic spines and development
As we have seen in the Cajal’s drawing (Fig. 6A), dendritic
appendages often curve and divide. This characteristic is easier
to see during development, when there is more space in the
neuropil for the movement of the appendages. The motility of
the spines decreases in adulthood. In contrast, filopodia are
highly motile structures that appear and disappear in minutes
(Dailey and Smith, 1996), and that are present during
development or in pathological states.
According to Dailey and Smith (1996) the filopodia lack a
differentiated head. In contrast, Fiala et al. (2002) propose that
filopodia can exhibit a bulbous head, where a synapse is
probably localized. Such a filopodium is still distinguishable
from a spine because it is longer than 2 mm and it is filled with a
dense matrix of actin (Fiala et al., 2002). In addition, filopodia
frequently ramify and curve (Morest, 1969; Fiala et al., 1998;
Dailey and Smith, 1996), like the appendages of Cajal’s
drawing (Fig. 6A).
Analyzing other Cajal drawings of developing cerebellum
(Fig. 7A and B), the two principal roles attributed to
filopodia can be seen (for a review, see Yuste and Bonhoeffer,
2004):
126 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130

Fig. 11. Different models for spinogenesis: (A) spines emerge independently of the afferent axon; (B) first synapses are formed on the dendritic shaft induced by the
axon terminal. At the beginning most of the spines are ‘‘stubbies’’. In the last stage many spines are mushroom-shaped or lollipop-shaped. (C) A dendritic filopodium
captures an axonal terminal and becomes a spine. The diagram has been created with fragments of 3D-reconstructions of neurons from Cajal’s histological
preparations. The design has been made following the diagram of Yuste and Bonhoeffer (2004).

- spinogenesis (Dailey and Smith, 1996) and synaptogenesis
(Harris et al., 1992);
- dendritic tree branching (Morest, 1969).
Two different populations of filopodia probably exist
(Portera-Caillau et al., 2003): those of the tip of the dendritic
branch, implicated in branching and modeling of the
dendritic tree; and those distributed along the dendritic
branch, implicated in synaptogenesis (Yuste and Bonhoeffer,
2004).
Although the work of Dailey and Smith (1996) suggests a
role of dendritic filopodia in spinogenesis, it is still not clear
how dendritic spines are formed. Three models have been
proposed: The Sotelo model (1978), the Miller/Peters model
(1981), and the filopodia model (Yuste and Bonhoeffer, 2004)
(Fig. 11).
Konur and Yuste (2004) have confirmed that the presynaptic
terminal is also motile and that the filopodia of the dendrite can
make transient interactions with them indicating that the
filopodia are implicated in sampling the surrounding neuropil
looking for a presynaptic terminal. In contrast, spines and hand-
like spines are less motile and their movement (head morphing)
may be implicated in synaptic competition between two
presynaptic terminals.
As we have seen, the genius of Cajal was not only to describe
the structures he was observing, but also to correctly interpret
them. Cajal also applied his neurotropic hypothesis to
development, especially that of the cerebellum, but moreover
he pointed out the importance of nervous activity in the
maturation of the dendritic tree (Cajal, 1899a,b). In the ideas of
Cajal the basis for the synaptotropic theory of Vaughn (1989)
can be found in the words of Cajal:
‘‘It appears, therefore, that the innumerable processes and
intercellular connections offered by the adult nervous
system can be interpreted as the morphologic expression
of the innumerable routes drawn up in the space by currents
of inducting or positive chemotropic substances during the
entire developmental period. Thus, the total arborization of a
neuron represents the graphic history of conflicts suffered
during its embryonic life’’.30 (Cajal, 1899a,b).
6. Conclusion
Dendritic spines are among the most outstanding topics of
research in present-day neuroscience. The concept of dendritic
spines has evolved in the past century since they were
discovered by Cajal. Technological advances, from electron
microscopy to two photon microscopy, have permitted a change
of the concept of these structures as key elements in
connectivity and synaptic plasticity. However, many of the
recent ideas about dendritic spines were first proposed by Cajal
and other scientists of his time.
Acknowledgement
The authors are supported by The Ramón Areces Founda-
tion.
30
‘‘Por donde se ve que el sinnúmero de expansiones y conexiones inter-
celulares ofrecidas por el sistema nervioso adulto, cabe concebirse como
expression morfológica de los innumerables caminos trazados en el espacio
y durante todo el perı́odo evolutivo, por las corrientes de las materias reclamo.
La arborización entera de una neurona representa, pues, la historia gráfica de
los conflictos sufridos durante la vida embrionaria’’.
 P. Garcı́a-López et al. / Progress in Neurobiology 83 (2007) 110–130
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