Microscopic Anatomy

11/11/09
Mary Ann Stepp, Ph.D.
Ross Hall, Rm. 226C, x4-0557
mastepp@gwu.edu

The Eye
Functions of the eye: As one of the
organs of the special senses, the eye
functions both to acquire and to begin
the initial processing of visual
information. The cornea and lens focus
light on the retina and the retina
integrates and sends the information to
the visual cortex of the brain. The
demand for the processing of visual
information by the brain is one of the
major forces behind its evolution into
the complex structure it is in humans.

The human eye consists of 3 primary

layers.
1. Inner: the retina-- both a
photosensitive and a non-
photosensitive part.
2. Middle: uvea or vascular tunic and
consists of the choroid, ciliary
body and process, zonule fibers,
and iris.
3. Outer: fibrous tunic or tunica
fibrosa and includes the lamina
cribosa, the sclera, trabecular
meshwork, canal of Schlemm,
corneoscleral junction (limbus),
and the cornea.

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And the eye has 3 major
compartments. The main
cavity of the eye behind the
lens is called the vitreous
space and is filled with the
vitreous humor, a transparent
gelatinous substance. The
area behind the cornea but in
front of the iris is called the
anterior chamber. The area
behind the iris and in front of
the lens is called the
posterior chamber. Both
anterior and posterior
chambers are filled with
aqueous humor.

Development

The optic sulcus forms as a depression in

the developing forebrain. It continues to
enlarge and it becomes termed the optic
vesicle. A region in the overlying
ectoderm begins to thicken and is then
termed the lens placode. As the lens
placode continues to thicken and the
optic vesicle enlarge, they come in
contact. Once they touch, the cells of the
lens placode begin to invaginate forming
the lens vesicle and the optic cup--the
outer-most layer will become the retinal
pigment epithelium (RPE) and the inner-
most layer the neural retina. The lens is
finally pinched off completely. The
cornea will form from the overlying
ectoderm. Thus, both lens and cornea are
derived from the ectoderm,

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The Inner Layer:
Has 2 major
components…
1.  photosensitive retina
2. non-photosensitive
retina

The Inner Layer

The inner layer of the eye

consists of a neural
photosensitive aspect called
the retina located in the
posterior globe and a two cell
layer anterior aspect, starting
at the ora serrata and
continuing over the ciliary
body and processes and over
the posterior aspect of the iris.
Here, after a brief mention of
the retinal pigment
epithelium, we will discuss
the photosensitive (neural)
retina.

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 The retinal pigment
epithelium (RPE): The
basal membrane of the RPE
abuts Bruch's membrane
and the choriocapillary
layer of the middle layer of
the eye called the choroid.
The apical membrane has
extensive invaginations:
microvilli and cylindrical
sheaths. These projections
are not anatomically sealed
by any specialized junctional
complexes.

The functions of the RPE

include:
1)  transport of Vit A to
the photoreceptors
2)  phagocytosis of
discarded rod/cone
outer segments
3)  absorb excess light
after photoreceptors
have been stimulated,
4)  support attachment of
the photoreceptors.

Note: each RPE cell likely phagcytoses 7,500 shed disc’s per day

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 Clinical Relevance 1: Retinitis
Pigmentosum(RP)
•  Genetic mutations in various
genes cause damage to the
RPE reducing the ability of
the RPE to remove shed rod
and cone outer segments and
resulting loss of rods and then
cones. There are over 100
different mutations to date
have been found that lead to
RP. They fall in all three
categories: autosomal
dominant, autosomal
recessive, or X-linked
inheritance and the incidence
is believed to be 1:4000.

Clinical Relevance 2: Retinal

Detachment
•  The neural retina detaches
from the RPE cells and the
choroid and the rods and cone
begin to starve. Patients
suddenly see lots of floaters
and can see flashes of light or
what seems like a veil
interfering with their vision.
•  If not repositioned quickly,
the rods and cones die and
fibroblast-like cells grow over
the RPE. Lasers are used to
reattach the retina.

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 Photosensitive retina is complex consisting of over 15
types of neurons including the rods and cones, the
bipolar cells and the ganglion cells. The retina has an
inverted structure: light initially goes past the neurons
without exciting them. Only after excitation of the
photoreceptors in the inner most layers are signals
transmitted back to the bipolar and ganglion cells.

The layers of the retina are distinct in

histological sections. The (1) internal
or inner limiting membrane serves as a
kind to protective basement membrane
for the ganglion cells. It overlies the (2)
ganglion cells which are above a synaptic
layer where the ganglion cells make
contact with the underlying bipolar cells.
This layer is called the (3) internal or
inner plexiform layer. Next comes the
(4) internal or inner nuclear layer
where the nuclei of the bipolar cells are
located. Another synaptic layer is next
where the bipolar cells contact the
photoreceptor cells and this layer is called
the (5) external or outer plexiform
layer.

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The photoreceptor (rod and cone) nuclei
are located next in the (6) external or
outer nuclear layer. The photoreceptors
must pass through (7) the external or
outer limiting membrane next. This is
where the cells interact with junctional
processes formed by specialized glial cells
called Mueller cells. The next layer visible
is the layer containing (8) the rod and
cone inner segments, then (9) the rod and
cone outer segments, and finally the outer
segments interdigitate with the (10) RPE.
The RPE lie over Bruch's membrane and
the choriocapillary layer of the choroid.

Rods and cones are

polarized neurons.
There are ~120 million
rods in the human eye
and only 6 million
cones. Rods function
in low levels of light,
and have no capability
of differentiating
among colors. Cones
function in daylight.
There are 3 types of
cone cells.

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Cones come in three types each of which is
“tuned” to a different wavelenth of light: red,
green, and blue. The cells contain one of three
visual pigments, called photopsins. As a result,
humans see in trichromy. Some people have
mutations that shift the wavelength optima in
their photopsins but they still have 3 types of
cone cells; others have just two types of cone
cells since they have mutations that prevent the
formation of one of the 3 photopsins.

Ishihara Plates

http://www.toledo-bend.com/colorblind/Ishihara.html

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 The density of rods and cones is not
uniform across the globe; some
parts of the retina are more sensitive
to light than others because of
differences in the distribution of
neurons.

In bright light, the contraction of the iris

allows the the cornea and lens to focus
the light on a region of the retina called
the fovea centralis (1.5 mm diameter)
which is located within a depression in
the retina called the macula (5.5.mm
diameter). The foveola (200 µm
diameter) is located within the fovea; it
is a where blood vessels are sequestered
to the periphery of the choroid layer.
The bodies of the ganglion and bipolar
cells are restricted to the periphery of the
fovea. The density of ganglion cells in
the at the fovea is much greater than any
where else in the retina. The macula is
also called the macula lutea and it
appears yellow in fundus photographs.
Also, the cone cells are tightly packed
within the macula and there are no rods.
This means that visual acuity and color
discrimination are optimal when light is
focused on the macula.

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In the TEM and SEM images shown, A and B
show the fovea and C and D regions at the
periphery of the retina. Images are shown in
two orientations.

Notice that in the fovea, there is only one type

of photoreceptor, cones, and the cones are very
much squished together and elongated
compared to cones in the periphery.

Clincal Correlation: Age-Related Macular

Degeneration (AMD) is the most frequent
cause of blindness in people over 65. 1.75
million people in the USA have it. Yellow
pigment (drusen) accumulates in and around the
macula and interferes with vision. The disease
has no treatment and progresses from a so-
called dry form (non-vascular) to a wet form
(vascular). Recently, progress have been made
trying to sort out the diseases genetics. Most
likely, macular degeneration is autoimmune in Dry AMD
nature and is made worse be smoking,
hypertension, and obesity.

Wet AMD

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Bipolar cell bodies are in the
internal nuclear layer; these
neurons integrate signals from
rods and cones and send the
signals on the the ganglion
cells. Bipolar cells form
junctions with the rods and
cones in the external
plexiform layer and with the
ganglion cells in the internal
plexiform layer.

The ganglion cells project

their axons to a specific
region of the retina where
they come together. This
site is called the optic
papilla or optic disk. The
bundle of axons is called
the optic nerve and this
site is also called the optic
nerve head. Because there
are no rods or cones here,
this is a "blind spot" in
the eye.

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Other cells in the retina include
horizontal cells which facilitate
interaction between photoreceptors,
amacrine cells which connect
ganglion cells and various types of
supporting cells including Mueller
cells. All these support cells have
their nuclei in the inner or internal
nuclear layer.

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 Mueller cells form the scaffold for
the entire retina. They span the
width of the neural retina. Both the
internal and external limiting
membranes are formed from the
Mueller cells. Mueller cells
therefore face the vitreous internally
and the subretinal space externally.
The superficial blood vessels within
the neural retina are also enfolded
by Mueller cells.
The Meuller cells are the primary
cells that make the vitreous. The
vitreous is a clear, gel-like substance
containing collagen and
proteoglycans present posterior to
the lens whose function appears to
be related to maintaining the shape
of the eye and in maintaining retinal
attachment.

The Middle Layer

(Uveal Tract)
Also has 4 major parts…
1. Choroid
2. Ciliary Body
3. Ciliary Processes
4. Iris

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1. Choroid: Primarily functions as
vascular support for the neural retina and
other ocular structures. The melanocytes
help to absorb any light not absorbed by
rods and cones of the neural retina.
Consists of loose connective tissue,
leukocytes, fibroblasts, melanocytes, and
the endothelial cells comprising the vessel
walls. The choroid has an outer and inner
layer. The outer layer has no specific
name. The inner layer is called the
choriocapillary layer and it is richer in
smaller vessels than is the outer layer. A
membrane separating the choriocapillary
layer from the retina is called Bruch's
membrane; the choriocapillary layer and
Bruch’s membrane are present from the
ora serrata to the optic disk.

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2. Ciliary Body: At the level of the lens,
the choroid becomes the ciliary body. The
ciliary body is triangular and contacts on
one side the vitreous humor, on another the
sclera, and on the third side it contacts the
posterior chamber (area between iris and
lens). The ciliary muscles are within the
ciliary body. The surface of the ciliary body
is lined with a 2 cell layer, non-
photosensitive, anterior extension of the
retina. The outermost cells are an extension
of the RPE cells and are melanin rich. The
innermost cells are derived from the neural
retina and are a simple columnar epithelium.
The epithelial cells of the ciliary body and
processes produce the aqueous humor.

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3. Ciliary Processes: Ridge-like
protrusions arising from the
ciliary body which are also
covered on their surface by the
non-photosensitive aspect of the
anterior retina. From the ciliary
process emerge the ciliary zonules
which insert into the lens capsule
to keep the lens in place and to
allow the lens to change shape
during accommodation. The cells
which make up the internal aspect
of the ciliary process transport
nutrients from the plasma through
vessel walls into the posterior
chamber and are thus responsible
for the formation of the aqueous
humor.

3.  Ciliary Processes,

cont’d
The aqueous flows
from the posterior
chamber, past the iris,
through the pupil, and
into the anterior
chamber to the
limbus where it
passes through the
trabecular meshwork
and into the canal of
Schlemm.

Red arrows indicate direction of

aqueous humor flow

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3.  Ciliary Processes,
cont’d

Clinical Correlation:
Glaucoma is caused by
blockage aqueous
outflow via Schlemm’s
Canal: Open vs Closed
angle glaucoma.

4. Iris: The function of the iris is to block stray light from entering the eye during daylight vision;
during night vision, the opening of the pupil by dilation of the iris allows for maximum light entry
into the eye. The iris is an extension of the choroid which lies anterior to the ciliary body and
processes. It partially covers the lens leaving a hole called the pupil. The anterior surface of the iris
is lined by mesothelial cells sitting on a basement membrane continuous with that of the posterior
surface of the cornea. Beneath this basement membrane is a stroma formed by fibroblasts and
melanocytes. The middle layer of the iris consists of circular and radiating muscle bundles which
interact closely with the posterior aspect of the iris. The posterior surface of the iris is lined by the 2
cell layer non-photosensitive aspect of the neural retina. This part of the iris is heavily pigmented
and has a purple hue.

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Summary:
The Middle Layer
(Uveal Tract)

1. Choroid:
2. Ciliary Body:
3. Ciliary Processes
4. Iris

The outer layer

Has 4 major parts…
1. Sclera
2. Lamina cribosa
3. Corneoscleral
junction (limbus)
4. Cornea

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The outer layer

1. The sclera: thick collagen rich tunic to

support the globe of the eye. Allows the
eye to achieve the high intraocular fluid
pressures necessary to maintain its shape.
By maintaining its shape, the eye can
provide stable refractive surfaces
necessary to permit focusing of light on
the retina. Animal models studying the
causes of myopia show that you can
induce myopia by occluding one of the
eyes during certain critical stages of eye
deveopment. When the sclera grows too
much, myopia develops.

The anterior aspect of the sclera at the

level of the ora serrata is covered by a
specialized epithelial cell layer called the
conjunctiva which is continuous with the
cells lining the inner aspect of the eyelids.

2. Lamina cribosa:
specialized area of the
sclera which helps to
protect and support the
optic nerve as it leaves
the eye. Located
beneath the optic papilla
or optic nerve head.
This is the structure the
ophthalmologist views
when looking for early
signs of glaucoma; it is
sensitive to increased
intraocular pressure and
begins to show signs of
breakdown in the
diseased eye.

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 OD= optic disk, BV=blood vessels, LC= lamina cribosa, ON= optic nerve

3. Corneoscleral junction
(limbus): region where the sclera
and cornea meet. The limbal
epithelial layer contains
specialized stem cells which serve
as the source of the cells of the
central corneal epithelium. This
region also functions in
maintaining intraocular pressure
since within the stroma beneath
the limbus lies the trabecular
meshwork and the canal of
Schlemm. The aqueous humor
drains into these structures to exit
the eye. Blockage of this
drainage system causes elevated
intraocular pressure, a condition
called glaucoma. Glaucoma left
untreated will cause damage to
the optic nerve, severe ocular
pain, and lead to detachments of
the retina and blindness.

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4. Cornea: The transparent, anterior
1/6th of the sclera. The function of
the cornea is to provide a clear
refractive surface to allow light into
the eye; its tough surfaces protect the
inner neuronal tissues from injury.
The curved shape allows light to be
focused on the retina. The cornea has
several layers: An outer stratified
squamous epithelial cell layer which
sits on a specialized acellular
basement membrane, called
Bowman's layer. The middle layer
or corneal stroma is lined above by
Bowman’s layer and below by
another specialized basement
membrane called Descemet's
membrane. The inner layer of the
cornea is composed of the corneal
endothelial cells which interact with
Descemet's membrane above and
with aqueous humor below.

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The epithelial cells of the cornea
have two principle functions: to
provide a smooth surface on
which the tear film may spread
and to act as a barrier to block
microbial entry into the inner
layers of the eye. These cells are
similar in many ways to the cells
which make up the epidermis but
are also quite unique in that they
have a surface designed to
promote tear film spreading, they
do not have a vascular source, and
they do not differentiate from
stem cells located in their basal
cell layer. In response to injury,
these cells migrate quickly to
cover exposed basement
membranes and stroma.
The corneal epithelium can
become hazy after surgery
(LASIK, PRK) and can detach
after healing in some patients
with recurrent erosions.

Human Corneas

After PRK Recurrent Epithelial

Erosion

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 In contrast, in skin, the basal cells layer, called stratum
germanitivum contains the stem cells and is not smooth
and refractive but undulating.

5. The stromal fibroblasts produce

the collagen molecules and
proteoglycans which form the
ordered arrays of collagen bundles
necessary to maintain both corneal
clarity and strength. These cells also
are important in the healing process
since they produce the matrix
degrading enzymes required to
degrade defective collagen fibers. In
response to injury these cells can
convert from a fibroblast phenotype
to cells which act like
myofibroblasts; they become motile
and help in wound contraction after
injury.

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6. The endothelial cells are
essential to the health of the cornea.
They are the cells which pump
nutrients from the aqueous humor
and release them into the stroma
where they passively diffuse to feed
the stromal fibroblasts and the
epithelium. The endothelial cells
also act as pumps to regulate the
flow of water into and out of the
cornea. They respond rapidly to
changes in the water content of the
cornea. Unfortunately, in adult
humans, the endothelial cells do not
replicate and over the lifetime of an
individual, they continually decrease
in number. Injury to these cells was
common during the early days of
cataract extraction procedures and
caused many corneas to become
cloudy and opaque requiring the
need for corneal transplantation.

Cornea Review:

The cornea is the most frequently transplanted organ. "Immune privileged" is a term frequently
used when referring to the cornea since corneal transplants are usually performed without tissue
typing.

The cornea functions primarily in vision to refract light. Changes in corneal shape can alter this
refractive power. For many people, their cornea is not the optimal shape to allow light to focus
on the retina properly. This is usually easily corrected by wearing glasses, contact lens, or by
surgical intervention. eg.LASIK.

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The lens has a thick capsule which covers the entire outer surface. The capsule is collagen-
rich, refractive, elastic, and is the site where the ciliary zonules attach to hold the lens in place
and change its shape. At the anterior aspect of the lens, beneath the capsule, there is a single
layer of cuboidal epithelial cells, called the subcapsular epithelium. As the lens ages, new cells
are derived from the cells at the equator of this region. Cells leave the equator of the epithelial
layer and elongate, become post-mitotic, and fill up with proteins called crystallins and
become known as lens fibers. Their nuclei shrink and RNA synthesis stops. The center of the
lens is called the nucleus or medulla; the outer layer is called the cortex. Both nucleus and
cortex are filled with lens fibers. The nearer to the nucleus, the older the fibers are because the
fibers are laid down in layers. The lens fibers in the nucleus of the lens contain crystallin
proteins synthesized within the embryo.

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 The lens functions in accommodation, that is the
ability to focus light on the retina from both near
and far objects. The lens does this by changing its
shape. The shape changes are controlled by the
muscles of the ciliary body and processes. At birth
the lens is highly elastic and can change shape
readily. As it ages, it becomes increasingly more
rigid resulting in decreased accommodation, a
condition termed presbiopia. In addition to losing
elasticity as it ages, the lens also begins to become
less transparent. This cloudiness is caused by
precipitation and coagulation of proteins within the
lens fibers and is called a cataract. It can also
result from injury to the lens. Precipitation of
proteins within the lens is speeded up by elevated
exposure to UV light, by elevated mineral levels
(Mg2+, Ca2+) and by elevated glucose (eg. more
common in diabetics). Removal of cataracts and
insertion of artificial intraocular lenses is highly
successful.
The mammalian lens is optically a better lens
than anything man can make.

Associated structures

1). Conjunctiva: bulbar

(covers anterior aspect of the
eye up to the corneal limbus)
and palpebral (lines the inner
surface of the eyelid) conj
function as barriers and contain
goblet cells which secret
mucins which help promote the
spreading of the tear film over
the corneal surface.

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2). Eyelids: The inner eyelid is lined by the conjunctiva; the outer surface by a thin epidermis. Within
the tarsal plate in connective tissue adjacent to the conjunctiva are the Meibomian Glands. Eyelashes
arise at the conjunctival: tarsal border with glands of Zeiss near their base and the gland of Moll nearby.
The orbicularus oculi muscles are also present near the midline of the eyelid; these muscle participate
in opening and closing the lid. The primary function of the blink is to distribute the tear film over the
eye surface; a secondary function is to prevent injury to the eye. All of the glands, and including the
lacrimal glands which are located in the anterior superior temporal orbit of the eye and the goblet cells
of the conjunctiva, participate in maintaining a moist ocular surface.

2). Eyelids,
continued…
Conditions which inhibit
tear production and/or
the blink reflex are
serious and cause a
condition called dry eye.
Dry eye allows corneal
epithelial denudation to
occur which then allows
bacteria to adhere and
infections to occur.
Scaring from these
infections eventually
interferes with vision.

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Review:

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Review:

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For answers to questions consult the
following sources:

Mary Ann Stepp

mastepp@gwu.edu Ross 226C
Or:
Histology:A Text and Atlas, Ross, et al., 4th
edition,2003.
Basic Histology, Junqueira, et al., 10thth
edition, 2003.
Grey's Anatomy, 30th American Addition
Molecular Biology of the Cell, 3rd
Addition, Alberts, et al., 1994
Principles and Practice of Ophthalmology,
Volume 1: Basic Sciences, Alberts and
Jacobiac et al., 1994.
The Cornea, Smolin and Thoft, 1994.

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