Biología de la Diferenciación Celular

Tema 4. Desarrollo de la especificación celular.

Concepto y mecanismo de determinación,

especificación y diferenciación celular

Desarrollo de la especificidad celular. Concepto y mecanismos
de determinación, especificación y diferenciación celular

1. Conceptos: totipotencia, pluripotencia, unipotencia

2. Compromiso celular

3. Determinación celular

4. Especificación celular. Tipos

Especificación autónoma

Especificación condicional

Especificación sincitial

5. Diferenciación celular

6. Morfogénesis
 Conceptos básicos

Totipotencia (totus: entero, completo)

Æ Oocito fertilizado o cigoto

Æ Potencial de generar todos los tejidos y células que forman el

embrión mediante procesos de división y especialización celular

Pluripotencia (plures: varios, muchos)

Æ Células que originan células derivadas de las tres capas germinales
embrionarias (ectodermo, mesodermo, endodermo)

Unipotencia (unus: uno)

Æ Células capaces de diferenciarse en un solo linaje
¿Cuándo pierden su totipotencia las células individuales del embrión en
desarrollo?

¿En qué momento del desarrollo las células individuales que formarán, por
ejemplo, el corazón, se distinguen del resto?

¿Cuándo adquieren el potencial restringido de desarrollo los descendientes

del oocito fertilizado?

Esta determinación: ¿es un proceso irreversible?

¿Las células especializadas que formarán los órganos provienen de una

única célula o de un grupo de ellas?

¿Existen “decisiones” que dirigen a las células embrionarias tempranas por

diferentes vías?

Los límites del crecimiento y la forma de las distintas partes del cuerpo:
¿cómo se establecen?
Diferenciación
Æ Desarrollo de tipos celulares especializados

Æ Proceso por el que los genes se expresan selectivamente y la acción de los

productos génicos da lugar a una célula con fenotipo especializado

Æ Cambios en la morfología, bioquímica, metabolismo y función celulares

Compromiso
Æ “Decisión” de una célula de entrar en el proceso de diferenciación a un cierto
destino

Æ Destino restringido en el desarrollo

Æ Fases:

Especificación: la célula es capaz de diferenciarse autónomamente en un

medio ambiente neutral

Determinación: la célula es capaz de diferenciarse autónomamente

incluso si se la coloca en otra región del embrión
 Algunos tipos celulares diferenciados

Tipo celular Producto Función especializada

Queratinocito Queratina Protección (abrasión, desecación)

Eritrocito Hemoglobina Transporte de oxígeno
C. del cristalino Cristalinas Transmisión de la luz
Linfocitos B Inmunoglobulinas Síntesis de anticuerpos
Linfocitos T Linfoquinas Destrucción células extrañas
Melanocitos Melanina Producción de pigmento
C. islotes pancreáticos Insulina Metabolismo carbohidratos
C. de Leydig Testosterona Caracteres sexuales masculino
Condrocito Condroitín-S, colágeno Tendones y ligamentos
Osteoblastos Matriz ósea Soporte esquelético
Miocitos Actina y miosina Contracción muscular
Hepatocitos Albúmina, enzimas Proteínas séricas y enzimas
Neuronas Neurotransmisores Transmisión impulso eléctrico
 Tipos de especificación celular

1. Especificación autónoma
Æ Desarrollo en mosaico
Æ Determinantes morfogenéticos

2. Especificación condicional
Æ Interacción con células vecinas
Æ Regulación: Desarrollo regulativo
Æ Regeneración: Gradientes de morfógenos
Æ Campos morfogenéticos

3. Especificación sincitial
Æ Interacciones entre distintas áreas de una célula
Modes of cell type specification and their characteristics

I. Autonomous specification
Characteristic of most invertebrates
Specification by differential acquisition of certain cytoplasmic molecules present in the egg
Invariant cleavages produce the same lineages in each embryo of the species
Blastomere fates are generally invariant
Cell type specification precedes any large-scale embryonic cell migration
Produces "mosaic" ("determinative") development: cells cannot change fate if a blastomere is lost

II. Conditional specification

Characteristic of all vertebrates and few invertebrates
Specification by interactions between cells
Relative positions are important
Variable cleavages produce no invariant fate assignments to cells
Massive cell rearrangements and migrations precede or accompany specification
Capacity for "regulative" development: allows cells to acquire different functions

III. Syncytial specification

Characteristic of most insect classes
Specification of body regions by interactions between cytoplasmic regions prior to cellularization of
the blastoderm
Variable cleavage produces no rigid cell fates for particular nuclei
After cellularization, conditional specification is most often seen
Gilbert, 2003
 Tipos de especificación celular

1. Especificación autónoma

Æ Desarrollo en mosaico

Æ Determinantes morfogenéticos: especificación del tipo

celular
 Especificación autónoma (desarrollo en mosaico)

Estadío de 16 Estadío de 48 Larva

células células

Figure 3.7. Autonomous specification (mosaic development). (A-C) Differentiation of trochoblast (ciliated) cells of the mollusc Patella. (A) 16-cell stage seen from the
side; the presumptive trochoblast cells are shaded. (B) 48-cell stage. (C) Ciliated larval stage, seen from the animal pole. (D-G) Differentiation of a Patella trochoblast cell
isolated from the 16-cell stage and cultured in vitro. (E, F) Results of the first and second divisions in culture. (G) Ciliated products of (F). Even in isolated culture, the cells
divide and become ciliated at the correct time. (Gilbert, 2003).
 Especificación
autónoma

Embriones de tunicado

Figure 3.8. Autonomous specification in

the early tunicate embryo. When the
four blastomere pairs of the 8-cell embryo
are dissociated, each forms structures that
it would have formed if it had remained in
the embryo. (The fate map of the tunicate
shows that the left and right sides produce
identical cell lineages.) (Gilbert, 2003).
 Especificación
autónoma

Expresión de acetilcolinesterasa
en la progenie de las blastómeras
de la línea muscular Acetilcolinesterasa Acetilcolinesterasa
(músculos de la cola) (progenie blastómera B4.1)

Embriones de tunicado

Figure 3.9. Acetylcholinesterase in the progeny

of the muscle lineage blastomeres (B4.1)
isolated from a tunicate embryo at the 8-cell
stage. (A) Diagram of the isolation procedure. (B)
Localization of acetylcholinesterase in the tail
muscles of an intact tunicate larva. The presence
of the enzyme is demonstrated by the dark
staining. The same dark staining is seen in the
progeny of the B4.1 blastomere pair (C), but not
in the remaining 6/8 of the embryo (D) when
incubated for the length of time it normally takes
to form a larva. (Gilbert, 2003).
Resto de la larva
 Tipos de especificación celular

2. Especificación condicional

Æ Interacción con células vecinas

Æ Regulación: Desarrollo regulativo

Æ Regeneración: Gradientes de morfógenos

Æ Campos morfogenéticos
 Especificación
condicional

Interacción con células vecinas

Regulación: Desarrollo regulativo

Figure 3.11. Conditional specification.

(A) What a cell becomes depends upon its
position in the embryo. Its fate is
determined by interactions with
neighboring cells. (B) If cells are removed
from the embryo, the remaining cells can
regulate and compensate for the missing
part. (Gilbert, 2003).
 Especificación condicional en el armadillo de nueve
bandas (Dasypus novemcinctus)

Figure 3.12. In the early developmental stages of many vertebrates, the separation of the embryonic cells into two parts can
create twins. This phenomenon occurs sporadically in humans. However, in the nine-banded armadillo, Dasypus
novemcinctus, the original embryo always splits into four separate groups of cells, each of which forms its own embryo.
(Gilbert, 2003).
Especificación Desarrollo regulativo
condicional

Erizo de mar

Figure 3.15. Driesch's demonstration of

regulative development. (A) An intact 4-
cell sea urchin embryo generates a normal
pluteus larva. (B) When one removes the
4-cell embryo from its fertilization
envelope and isolates each of the four
cells, each cell can form a smaller, but
normal, pluteus larva. (All larvae are
drawn to the same scale.) Note that the
four larvae derived in this way are not
identical, despite their ability to generate
all the necessary cell types. Such variations
are also seen in adult sea urchins formed in
this way. (Gilbert, 2003).
Gradientes de morfógenos: Regeneración de las planarias

Figure 3.18. Flatworm regeneration and its limits. (A) If a flatworm is cut in two, the
anterior portion of the bottom half regenerates a head, while the posterior of the upper half
regenerates a tail. The same tissue can generate a head (if it is at the anterior portion of the
tail piece) or a tail (if it at the posterior portion of the head piece). (B) If a flatworm is cut
into three pieces, the middle piece will regenerate a head from its anterior end and a tail
from its posterior end. (C) However, if the middle piece is too thin, there is no morphogen
gradient within it, and regeneration is abnormal. (D) If the second cut is delayed, an
equally thin middle section forms a normal worm, since the time lag has allowed an
anterior-posterior gradient to become established. (Gilbert, 2003).
 Función de la activina
Cambios en la expresión génica

Brachiury

Goosecoid

Figure 3.20. Activin (or a closely related compound such as

Nodal) is thought to be responsible for converting animal
hemisphere cells into mesoderm. When animal cap cells were
removed from Xenopus blastulae and placed in saline
solutions containing activin, the activin conferred different
fates on the cells at different concentrations. (Gilbert, 2003).
 Efectos del gradiente de activina

Figure 3.21. A gradient of activin causes different gene expression in Xenopus animal cap cells. The mRNAs from the Brachyury and
goosecoid genes can be monitored by hybridization techniques that will be discussed in the next chapter. The cells containing these mRNAs
appear darker than the cells not expressing them. Beads containing activin induce the expression (transcription of mRNA) of the Brachyury gene
at distances removed from the beads. (A) Beads containing no activin did not elicit Brachyury gene expression. (B) Beads containing 1 nM
activin elicited Brachyury expression near the beads. (C) Beads containing 4 nM activin elicited Brachyury gene expression several cell diameters
away from the beads. However, goosecoid expression is seen where the concentration of activin is higher, near the source. Thus, it appears that
Brachyury gene expression is induced at particular concentrations of activin, and that goosecoid is induced at higher concentrations of activin.
(Gilbert, 2003).
 Campos morfogenéticos
Campo de la extremidad de la salamandra
Ambystoma maculatum

Figure 3.23. Prospective

forelimb field of the
salamander Ambystoma
maculatum. The central area
contains cells destined to
form the limb per se (the
free limb). The cells
surrounding the free limb
give rise to the peribrachial
flank tissue and the shoulder
girdle. The ring of cells
outside these regions usually
is not included the limb, but
can form a limb if the more
central tissues are extirpated.
(Gilbert, 2003).
 Tipos de especificación celular

3. Especificación sincitial

Æ Interacciones entre distintas áreas de una célula

 Especificación sincitial

Interacciones entre distintas áreas

de una célula

D. melanogaster

Figure 3.25. Syncytial specification in the fruit fly

Drosophila melanogaster. Anterior-posterior
specification originates from gradients within the egg
cell. Bicoid mRNA is stabilized in the most anterior
portion of the egg, while Nanos mRNA is tethered to
the posterior end. (The anterior can be recognized by
the micropyle on the shell; this structure permits
sperm to enter.) When the egg is laid and fertilized,
these two mRNAs are translated into proteins. The
Bicoid protein forms a gradient that is highest at the
anterior end, and the Nanos protein forms a gradient
that is highest at the posterior end. These two proteins
form a coordinate system based on their ratios. Each
position along the axis is thus distinguished from any
other position. When nuclear division occurs, each
nucleus is given its positional information by the ratio
of these proteins. The proteins forming these gradients
will activate the transcription of the genes that specify
the segmental identities of the larva and the adult fly.
(Gilbert, 2003).
 Morfogénesis

- Procesos que inducen cambios en la forma del organismo que se está

desarrollando

- Mecanismos muy diversos pueden conducir a cambios morfogenéticos

(invaginación, compactación, cavitación, ramificación, elongación,
migración)

- Depende de varias propiedades celulares:

- Adhesión celular

- Motilidad celular

- Comunicación celular
 Moléculas de adhesión celular

Figure 22-2. Major families of cell-adhesion molecules (CAMs). (Lodish et al., 1999).
 Estructura y función de las cadherinas

Figure 19-24. The structure and function of cadherins. (A) A classical cadherin molecule. The protein is a homodimer, with the
extracellular part of each polypeptide folded into five cadherin repeats. There are Ca2+-binding sites between each pair of repeats. (B)
The crystal structure of a single cadherin repeat, which resembles an immunoglobulin (Ig) domain. (C) The influence of extracellular
Ca2+. As the amount of Ca2+ increases, the extracellular parts of the cadherin chains become more rigid. When enough Ca2+ is bound,
the cadherin dimer extends from the surface, where it can bind to a cadherin dimer on a neighboring cell. If Ca2+ is removed, the
extracellular part of the protein becomes floppy and is degraded by proteolytic enzymes. (Alberts et al., 2002).
Estructura y función de las cadherinas

Figure 19-7. The cadherin

superfamily. (A) A classical cadherin
molecule. The protein is a homodimer,
with the extracellular part of each
polypeptide folded into five cadherin
repeats. There are Ca2+-binding sites
between each pair of repeats. (B) The
crystal structure of a single cadherin
repeat, which resembles an
immunoglobulin (Ig) domain. (C) The
influence of extracellular Ca2+. As the
amount of Ca2+ increases, the
extracellular parts of the cadherin chains
become more rigid. When enough Ca2+
is bound, the cadherin dimer extends
from the surface, where it can bind to a
cadherin dimer on a neighboring cell. If
Ca2+ is removed, the extracellular part
of the protein becomes floppy and is
degraded by proteolytic enzymes.
(Alberts et al., 2002).
Estructura y función de las cadherinas

Figure 19-9. The structure and function of cadherins. (A) A classical cadherin
molecule. The protein is a homodimer, with the extracellular part of each
polypeptide folded into five cadherin repeats. There are Ca2+-binding sites
between each pair of repeats. (B) The crystal structure of a single cadherin repeat,
which resembles an immunoglobulin (Ig) domain. (C) The influence of
extracellular Ca2+. As the amount of Ca2+ increases, the extracellular parts of the
cadherin chains become more rigid. When enough Ca2+ is bound, the cadherin
dimer extends from the surface, where it can bind to a cadherin dimer on a
neighboring cell. If Ca2+ is removed, the extracellular part of the protein becomes
floppy and is degraded by proteolytic enzymes. (Alberts et al., 2008).
 Major Cadherin Molecules on Mammalian Cells

Molecule Predominant Cellular Distribution

E-cadherin Preimplantation embryos, non-neural epithelial

tissue

P-cadherin Trophoblast (placenta)

N-cadherin Nervous system, lens, cardiac and skeletal

muscle

SOURCE: M. Takeichi, 1988, Development 102:639; M. Takeichi, 1991, Science 251:1451; H. Inuzuka et al., 1991, Neuron 7:69; and M. Donalies et al., 1991,
Proc. Nat'l. Acad. Sci. USA 88:8024.

(Lodish et al., 1999).

 Expresión de E y N-cadherina en el sistema
nervioso en desarrollo

E-cadherina N-cadherina
Ectodermo Tubo neural
Figure 19-12. The distribution of E-cadherin and N-cadherin in the developing nervous system. Immunofluorescence micrographs
of a cross section of a chick embryo showing the developing neural tube labeled with antibodies against (A) E-cadherin and (B) N-
cadherin. Note that the overlying ectoderm cells express only E-cadherin, while the cells in the neural tube have lost E-cadherin and
have acquired N-cadherin. (Alberts et al., 2008).
 Estructura de
N-CAM

Figure 19-31. The cell adhesion protein

N-CAM. (A) Four forms of N-CAM. The
extracellular part of the polypeptide chain
in each case is folded into five Ig-like
domains (and one or two other domains
called fibronectin type III repeats).
Disulfide bonds (red) connect the ends of
each loop that forms an Ig-like domain. (B)
A model for the homophilic interaction that
allows N-CAM to mediate cell-cell
adhesion. (Alberts et al., 2002).
 Estructura de
N- e I-CAM

Figure 19-20. The cell adhesion protein

N-CAM. (A) Four forms of N-CAM. The
extracellular part of the polypeptide chain
in each case is folded into five Ig-like
domains (and one or two other domains
called fibronectin type III repeats).
Disulfide bonds (red) connect the ends of
each loop that forms an Ig-like domain. (B)
A model for the homophilic interaction that
allows N-CAM to mediate cell-cell
adhesion. (Alberts et al., 2008).
 Integrinas

Figure 19-45. The subunit structure

of an active integrin molecule, linking
extracellular matrix to the actin
cytoskeleton.
(B) Some of the proteins that form focal
adhesions. The transmembrane
adhesion protein is an integrin
heterodimer, composed of an a and a b
subunit. Its extracellular domains bind
to components of the extracellular
matrix, while the cytoplasmic tail of the
b subunit binds indirectly to actin
filaments via several intracellular
anchor proteins. (Alberts et al., 2008).
 Integrinas

Actina Vinculina

Figure 19-12. Focal adhesions. (A) In these immunofluorescence micrographs, cells in culture have been labeled with antibodies against
both actin (green) and the intracellular anchor protein vinculin (red). Note that vinculin is located at focal adhesions, which is also where
bundles of actin filaments terminate at the plasma membrane. (B) Some of the proteins that form focal adhesions. The transmembrane
adhesion protein is an integrin heterodimer, composed of an a and a b subunit. Its extracellular domains bind to components of the
extracellular matrix, while the cytoplasmic tail of the b subunit binds indirectly to actin filaments via several intracellular anchor proteins.
(Alberts et al., 2002).
 Experimentos de reagregación celular

Figure 3.26. Reaggregation of cells from amphibian neurulae. Presumptive epidermal cells from pigmented embryos
and neural plate cells from unpigmented embryos are dissociated and mixed together. The cells reaggregate so that one
type (here, the presumptive epidermis) covers the other. (After Townes and Holtfreter 1955.) (Gilbert, 2003).
Implicación de la matriz extracelular en la
diferenciación celular
Células de Sertoli

A. Placa de cultivo

B. Placa de cultivo cubierta

con lámina basal

Figure 6.35. Role of the extracellular matrix

in cell differentiation. Light micrographs of rat
Sertoli testis cells grown for two weeks (A) on
tissue culture plastic dishes and (B) on dishes
coated with basal lamina. The two photographs
were taken at the same magnification, 1200×.
(From Hadley et al. 1985; photographs courtesy
of M. Dym.) (Gilbert, 2003).
 Implicación de la matriz extracelular
en la diferenciación celular
Tejido glandular mamario

A. División en placa de cultivo

Genes implicados en división celular ON
Genes implicados en diferenciación celular OFF
Laminina
B. Placa de cultivo cubierta con membrana basal (laminina)
Genes implicados en división celular OFF
Genes implicados en la inhibición de la división celular ON
Genes implicados en diferenciación celular ON

C. Epitelio secretor rodeado de membrana basal

Activación del gen que codifica para caseína

D. Epitelio secretor rodeado de membrana basal

Activación del gen que codifica para proteína del suero

Figure 6.36. Basement membrane-directed gene expression in mammary gland tissue.

(A) Mouse mammary gland tissue divides when placed on tissue culture plastic. Cell
division genes are on, and the genes capable of synthesizing the differentiated products of
the mammary gland (lactoferrin, casein, whey acidic protein) are off. (B) When presented
with basement membrane that contains laminin, the genes for cell division proteins are
turned off, while the gene inhibiting cell division (p21) and the gene for lactoferrin are
turned on. (C, D) Mammary gland cells wrap the basement membrane about them, forming
a secretory epithelium. The genes for casein and whey protein are sequentially activated.
(Gilbert, 2003).
 Sistemas de señalización célula-célula

Figure 15-2. General schemes of intercellular signaling in animals. (a - c) Cell-to-cell signaling by extracellular chemicals occurs over
distances from a few micrometers in autocrine and paracrine signaling to several meters in endocrine signaling. (d) Proteins attached to the
plasma membrane of one cell can interact directly with receptors on an adjacent cell. (Lodish et al., 2008).
 Arrastre celular

Figure 11.32. Cell crawling The crawling movements

of cells across a surface can be viewed as three stages
of coordinated movements: (1) extension of the leading
edge, (2) attachment of the leading edge to the
substratum, and (3) retraction of the rear of the cell into
the cell body. (Cooper, 2004).