[Ocular biochemistry (OPT 318)

By Dr. (Mrs.) Uwagboe P.N

O.D , M.sc(Ocular Health).MNOA.

THE CORNEA

The cornea is the clear front surface of the eye. It lies directly in front of the iris and pupil, and it
allows light to enter the eye. The cornea is the transparent front part of the eye that covers the
iris, pupil, and anterior chamber. Together with the lens, the cornea refracts light, accounting for
approximately two-thirds of the eye's total optical power. In humans, the refractive power of the
cornea is approximately 43 dioptres.

The cornea has unmyelinated nerve endings sensitive to touch, temperature and chemicals; a
touch of the cornea causes an involuntary reflex to close the eyelid. Because transparency is of
prime importance the cornea does not have blood vessels; it receives nutrients via diffusion from
the tear fluid at the outside and the aqueous humour at the inside and also from neurotrophins
supplied by nerve fibres that innervate it. In humans, the cornea has a diameter of about 11.5 mm
and a thickness of 500-600 μm in the center and 600–800 μm at the periphery. Transparency,
avascularity, the presence of immature resident immune cells, and immunologic privilege makes
the cornea a very special tissue.

LAYERS/STRUCTURE OF THE CORNEA

1. Corneal epithelium
2. Bowman's layer
3. Corneal stroma
4. Descemet's membrane
5. Corneal endothelium

CORNEAL EPITHELIUM
The corneal epithelium is composed fairly uniformly of 5–7 layers of cells. It is about 50
μ in thickness. The epithelium is uniform to provide a smooth regular surface and is made
up of non -keratinized stratified squamous epithelium. The epithelium is derived from
surface ectoderm between 5 and 6 weeks of gestation. The epithelium and the overlying
tear film have symbiotic relationship. The mucin layer of the tear film which is in the
direct contact with corneal epithelium is produced by the conjunctival goblet cells. It
interacts closely with the corneal epithelial cells' glycocalyx to allow hydrophilic
spreading of the tear film with each eyelid blink. The innermost is called the basal cell
layer which germinate (regenerative) locus of the epithelial cells implying that the basal
epithelial cells divide and differentiate into the upper layer of the corneal epithelium .The
second layer, the wing cells consist of polyhedral cells, is located between the most
superficial and inner layer of the corneal epithelium. The size of the epithelial cells
increases as they move to the superficial layer and th e outermost 1to 2 layers are called
superficial cells. It blocks passage of foreign material,such as dust,water,and bacteria into
the eye and other layers of the cornea and provide a smooth surface that absorbs oxygen
and cell nutrients from tears, then distributes these nutrients to the rest of the cornea.
BASEMENT MEMEBRANE
 The basement membrane of the corneal epithelium is located between the basal cell of the
epithelium and Bowman layer. This layer is made up of two sub layers-the superficial
lamona lucida and the deeper lamina densa. The basement membrane contains type IV
and vii collagens and glycoproteins such as laminin and fibronectin.it is important in
providing physical and biochemical cues to the overlying cells.

BOWMAN’S LAYER

This is a very thin (8 to 14 microns) and dense fibrous sheet of connective tissue that forms the
transition between the corneal epithelium and the underlying stroma.it is made of types i, iii and
iv collagens and proteoglycans. It is non regenerating layer .

STROMA LAYER
This middle layer of the cornea is approximately 500 microns thick, or about 90 percent
of the thickness of the overall cornea. It is composed of strands of connective tissue
called collagen fibrils. These fibrils are uniform in size and are arranged parallel to the
cornea surface in 200 to 300 flat bundles called lamellae that extend across the entire
cornea. The regular arrangement and uniform spacing of these lamellae is what enables
the cornea to be perfectly clear by transmitting 98% of incoming light .This condition is
maintained by minimal hydration of the corneal stroma created by the endothelium.
DESCEMENT MEMBRANE
This very thin layer separates the stroma from the underlying endothelial layer of the
cornea. Descemet's (pronounced "DESS-eh-mays") membrane gradually thickens
throughout life — it's about 5 microns thick in children and 15 microns thick in older
adults.
CORNEA ENDOTHELIUM
This is the innermost layer of the cornea. The back of the endothelium is bathed in the
clear aqueous humor that fills the space between the cornea and the iris and pupil. The
corneal endothelium is only a single layer of cells thick and measures about 5 microns.
Most of the endothelial cells are hexagonal (six-sided), but some may have five or seven
sides. The regular arrangement of these cells is sometimes called the endothelial
mosaic.The endothelium maintains stromal deturgescence by functioning both as a
 barrier to fluid movement into the cornea and an active pump that moves ions and draw
water osmotically,from the stroma into aqueous humor.

FACTORS NECESSITATING CORNEAL TRANSPARENCY

Basically, there are three factors that account for the transparency of the cornea.

-Anatomical factors

-Physiological factors

-Biochemical factors

ANATOMICAL FACTORS

1. Corneal epithelium.

Epithelium are closely packed, uniformity and regularity in arrangement, Homogeneity in

refractive index. Tight intracellular junction.

The epithelial cells are homogenous

Normal epithelium is transparent due to homogenicity of its refractive index. • Basal cells are
firmly joined laterally to other basal cells and anteriorly to the wing cells by desmosomes and
macule occludentes. These tight intercellular junction accounts for corneal transparency. • As
well as it resistance to flow of water, electrolytes and glucose(BARRIER FUNCTION).

2. PRECORNEAL TEAR FLIM plays important role in maintaining the transparency by

keeping epithelial surface smooth and providing high quality optical surface.

• Therefore, condition associated with tear film abnormalities and corneal epithelial abnormality
may result in loss of corneal transparency.

3. ARRANGEMENT OF STROMAL LAMELLAE • Two theories have been put forward to

explain the role of peculiar arrangement of stromal lamella in corneal transparency. 1. Maurice
theory. 2. Theory of Goldman et al.

Maurice theory states that corneal transparency is because of the uniform collagen fibrils which
are arranged in a regular lattice so that scattered light is destroyed by the mutual interference. •
He stated that as long as the fibrils are regularly arranged in a lattice, seperated by less than a
wavelength of light (4000-7000A) the cornea will remain transparent. • Loss of transparency will
result if this regular arrangement is altered by stromal oedema or mechanical stress.

Goldman and Benedek theory of 1968 after applying diffraction theory to the problem
concluded that the lattice arrangement is not a necessary condition for stromal transparency. •
Rather they postulated that the cornea is transparent because the fibrils are small in relationship
to the light and do not interfere with light transmission unless they are larger than one half a
wavelength of light (2000 A) However, theory of Maurice as well as that of Goldman et al fail to
explain the occurrence of rapid clouding of cornea associated with acute raise of intraocular
pressure and rapid clearing of the cornea with reduction of intraocular pressure.

4 .ENDOTHELIUM

Single layer ,homogenous,closely packed cells.it has deturgescence function

5 CORNEAL VASCULARIZATION •

The normal cornea is avascularised except at the limbus . It facilitates nutrition, transport of
systemic antibiotic and drugs. • Progressive vascularization, however is a harmful process as it
interfere with functional properties of cornea, especially its transparency.

PHYSIOLOGICAL FACTORS

1. Corneal hydration.

Normal cornea maintains itself in a state of relative dehydration (80%water content) which is
essential for corneal transparency.it is kept constant by

1. Factors which draw water in the cornea, like stroma swelling pressure(sp)and intraocular
pressure(iop).

2. Factors which prevent flow of water in the cornea

Mechanical barrier function of epithelium and endothelium

3. Factors which draw water out of cornea

-active pumping action of endothelium

There are factors that sustain the state of corneal hydration which is vital for vision. Corneal
hydric balance is necessary for its transparency. Hyper hydration of the cornea reduces its
transparency and the cornea becomes translucent. In normal conditions 80% of the cornea is
water. Cornea hydration is maintained by anatomical integrity of the endothelium and the
epithelium. These two layers acts as barriers .epithelial and endothelial cells prevent water from
entering the corneal stroma. the following are factors that maintain the correct amount of fluid in
the cornea.

FACTORS AFFECTING CORNEAL HYDRATION

A STROMA SWELLING PRESSURE (SP); Glycosaminoglycans (GAGs) or

mucopolysaccharides are long unbranched polysaccharides consisting of a repeating
disaccharide unit. Glycosaminoglycans are highly polar and attract water. They are
therefore useful to the body as a lubricant or as a shock absorber.The pressure exerted by
GAGs is 60mmhg. Electrostatic repulision of anionic charges on the GAG molecue
expands the tissue ,sucking in the fliud with equal but negative pressure called imbibition
pressure(IP).

IP=IOP-SP 17-60= -43mmHg

Negative imbition pressure draws out water from stroma.

INTRAOCULAR PRESSURE (IOP).is a force performed by the intraocular

fluids and maintains intraocular pressure. Increased iop causes an increased water content
in the cornea and decreases transparency

When iop exceeds SP i.e when IP becomes positive, cornea oedema result .it can occur
when there is high IOP and normal SP, as in acute glaucoma, normal iop and low SP ,as
in endothelial dystrophy.

B BARRIER FUNCTION OF EPITHELIUM AND ENDOTHELIUM

 Epithelium and endothelium are semipermeable in nature

 Function as barriers to excessive flow of water and diffusion of electrolytes into
the stroma
  Epithelium offers most resistance to flow of water.

- C HYDRATION ACTION OF THE ENDOTHELIUM

i Na/K ATPase pump system

The endothelium is more active than epithelium, pump are located in the basolateral
membrane of the endothelial cell, the cornea floats between two liquids (aqueous and tear
film), so a balance must exist to equally lose and gain water. Higher sodium (Na+) levels out
from the cornea tend to dehydrate it. Whereas higher Na+within the cornea tends to hydrate
it. The endothelium pumps water and Na+out from the stroma .This balance is maintained by
the metabolic pumps

Enzyme (Na+K+activated ATPase) mediate pump causes extrusion of the Na and and water
from the stroma and thus maintain corneal transparency.

ii Bicarbonate dependent ATPase present in endothelium are also reported to have role in
fluid /ion balance in the cornea.

iii. cyarbonic anhydrase.enzyme catalyses the conversion of CO2 and water into HCO3-
and H+,thus provides important source for HCO3- for endothelial pump.

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BIOCHEMISTRY OF THE CORNEA
Like all other living tissues the cornea must undergo metabolism. A whole series of
enzymes and coenzymes are available to oxidize metabolites into energy. Generally the
cornea is not very active metabolically. This is because it presents certain structural
peculiarities which must to a certain extent affect its metabolism.it is normally
avascularized except at the limbal area, secondly, it is bathed on its surface by fluids
which are constantly being replaced and on one side is consistently subjected to
evaporation.it must thus drive its metabolites by diffusional processes from the limbal
capillaries, aqueous humour and the tears.
The cornea must derive its oxygen from air through the epithelium. Carbohydrate
metabolism is mainly through glycolysis and tricarboxylic acid cycle which mainly takes
place in the epithelium. other possible pathways include the pentose phosphate pathway
etc.in the stoma in order to reduce waste a series of specific dehydrogenates are used .
Several other factors influences corneal metabolism.
-Minerals such as iron,copper,Zinc and manganese are present in the cornea which
influences many respiratory enzyme systems.
-Also several vitamins such as Riboflavin, vitamin A,B complexes all influences corneal
metabolism.
The lactic acid and pyruvic acids formed by the glycolytic and PPP are degraded step
wise and excess lactate are transferred to the tears.

CLINICAL EVALUATION OF THE CORNEA

The corneal surface can be evaluated directly by inspection of the cornea and its sheen.
Magnifers can be used. Also the Ophthalmolscope could be used. However, the slit lamp
biomicroscope is the most ideal which could utilize any of the following types of
illumination.
-Direct illumination-used to detect gross anomalies

-Scleral scatter illumination-transmit light within the cornea by total internal reflection,
light is obstructed by any opacity and lesions which becomes illuminated as the path of
internally reflected light beam is obstructed.

-Retroillumination-Helps detect fine epithelial and endothelial changes,Keratic

precipitates and small blood vessels.

CORNEAL PERMEABILITY
 This has to do with how xenobiotics gain entry into the cornea.of more importance to us
is how drug penetrate the cornea.Thus the efficacay of any topical drug depends on the
permeability of the drug.The permeability of such drug is contingent primarily on their
lipid and water solubility.The epithelium and endothelium have a hundred times the lipid
content of the stroma thus fat/lipid soluble drugs could penetrate.However,only water
soluble drugs can penetrate the stroma.Therefore topical drugs for the cornea can be
amphipathic i.e it should be both lipophilic and hydrophilic so as to penetrate all the
layers of the cornea.

FACTORS AFFECTING CORNEAL PERMEABILITY

-lipid solubility

-water solubility

-molecular size and structure of the drug

-ionic charge of the drug

-Altering the permeability characteristics of the eye

-Drug contact time

As an example, Flourescein a negatively charged ion will not penetrate the epithelium. This is
the basis of the fluorescein dye test.if the epithelium is intact fluorescein will not stain the cornea
while if the epithelium is damaged or absent it will stain the damaged area or the stroma
respectively.

THE CRYSTALLINE LENS

The crystalline lens is one of the refractive media of the eye.it has no blood vessels, nerves or
connective tissue. Its avascularity and transparency is similar to that of the cornea.it is positioned
behind the iris and due to its flexibility is the chief refractive medium of the eye. Apart from the
optical attribute of transparency, it is elastic and biconvex and enclosed in a capsule which
withers with age.it refracts light entering the eye through the pupil and focuses it on the retina.
The transparency is dependent on biochemical, physiologic and anatomical integrity which gives
it the functional basis of an optical tissue.
The crystalline lens possesses external surface and internal curvatures whose ability with exact
refractive indices induces the manifestation of clear images on the retina. The crystalline lens is
one of the simplest structures of the complicated mammalian physiognomy. This is because it is
made of one cell type called epithelial cell which grows continuous and are sequested throughout
life in an encasement of an elastic and transparent membrane called lens capsule.

Throughout the period of growth and decay, older and worn out cells are never discarded as
occurs in other tissues of the body, rather, they are compressed towards the center with newer
cells growing on them. The process of this type of cellular growth and compression is continuous
until degenerative changes set in. The transparency of the lens of the eye is therefore dependent
on the critical regularity of their fibres and the balance of their chemical consituents. The lens is
isolated because it must be free from contact with the surrounding structures which might
hamper changes in the curvatures of its surfaces and also as an organ for accommodation, it must
be able, to increase its dioptric power with some space.

EMBROLYOGY OF THE LENS

Before the earliest stages of formation of the lens, a sequence of embroyological development
occurs. For the vertebrates, the formation of the crystalline lens originates from the surface
ectoderm which is in contact with the optic outgrowth. During the earliest period of embryonal
development ,the lens enlargement is homogenous with the area covering the surface ectoderm at
the 4mm where the optic outgrowth are seen as rounded prominence on the lateroventral surface
of the forebrain.

BIOCHEMISTRY OF THE LENS

The human lens is the least hydrated organ of the body having 66% water and 33% solid most of
which are proteins .the lens cortex is more hydrated than the nucleus. lens hydration is
maintained by an active Na+ ion water pump that resides within the membranes of cells in the
lens epithelium and the lens fibre.

THE LENS AS AN OSMOMETER

The entire lens capsule acts as osmometer because it exudes the properties like swelling in
hypotonic media and dehydration in the hypertonic, thus it has the capacity to function as an
osmometer.the osmolarity of the humans lens is 302mOsm and equals the osmolarity of aqueous
humour.The cations(Na+,K+)have a concentration of 145mEg/L and
anioins(CL-,SO4-,ascorbate,biocarbonate and glutathione)50-60mEg/L. The deficit is made up by
acidic groups such as lens protein and glucoproteins.

The lens as an osmometer is necessary because increase in H20 breaks the lens fibres resulting to
microscopic vacuoles being created. Also , the H20 equilibrium is disrupted in the lens if the
concentration of osmotically active compounds(Na+,K+) increases inside the lens.

CHEMICAL COMPOSITION OF THE LENS

LENS-PROTEINS

The human lens contains the highest concentration of protein (33%) than any other tissue. These
proteins are synthesized in the anterior epithelium and equatorial region. The almost perfect
physiochemical arrangement of lens proteins in an optimum environment of water , electrolytes,
and sulphydryl molecules gives the lens its transparency. Among the lens proteins ,15% of them
are insoluble in H20and these form the Albuminoid fraction which actually is made up of
membrane bound proteins and aggregated crystallins. The remaining 85% is H20 soulble and are
made up of Alpha(ᾳ )beta(B ) and Gamma( ‫) ﻷ‬crystallins which are so classified according to
molecular weight.The alpha crystallins constitutes 3.5%,B crystallins constitutes 55% and the
gamma the slowest 10%.

LIPIDS: The lipids in humans lens are unique and differ markedly from those of other spieces.
lens lipids represent about 3-5% dry weight of the lens with cholesterol constituting about 50%
of lipids, followed closely by phospholipids,45% and glycolipids and ceramides 5%.The lipids
are major components of the lens fibre and any decrease in synthesis or impaired synthesis
results to lens damage and subsequently, opacities. As an indication of the significance of lipids,
if an anticholesterol agent is used prolongedly such as triparanol,a side effect is the manifestation
of cataract.

Also, the cholesterol-phospholipid ratio of human lens fibres membrane is highest among all or
organelle membranes, thus confirming the lens with resistance to structural deformation and
transparency.
ASCORBATE

The ascorbate level of the lens is higher than that of AH. This high concentration is necessary for
the lens to participate in oxidation-reduction reactions. Also it works with other vitamins,
enzymes, metabolites for the maintenance of the optical properties of the lens. The high
concentration of ascorbic acid is because it enters the eye from plasma in oxidized form and is
reduced by the lens.it is assumed that the oxidized form could pass through the blood aqueous
barrier into the lens.

ELECTROLYTES

The electrolytes and water content of the intact lens resembles that of an intact cell. for the ocular
fluids they are similar to those of the biological fluid.it is important to note that to maintain
electrolyte and water gradients ,the lens which is surrounded by fluids(AH and VH)has to
generate both chemical and electrical energy.chemical energy extrudes Na+ ions and water is
produced by ATP through glucose metabolism.

Energy dependent pumps-Na+k+ ATpase pump in K+ and extrudes Na+. This is done by this
enzyme to degrade ATP TO ADP plus energy which is used to power the pump.This pumping
system can be stopped by cardiac glucosides. This pumpimg activities is more pronounced in the
epithelial surface because the concentration of Na+K+ ATPase is greater in this area. When K+is
pumped into the lens and Na+ is pumped out at the anterior surface, a chemical gradient is
generated that stimulates diffusion of Na+ into the lens and K+out primarily through the
posterior surface .This process of active transport (pump)stimulating passive diffusion

(leak)is called the PUMP AND LEAK THEORY OF CATION TRANSPORT.

The major function of this transport system are

-Regulation of water content of the lens making the lens to be like a perfect osomometer this
prevent colloid osmotic swelling.

-Produces and maintains an electrical potential difference of about 70Mv between the lens and
media surroundings it.

-Promotes the proper physiochemical environment within the lens so as to maintain transparency
and optimal enzymatic activity.
Surface active agents(antiobiotics, detergents, lysophopholipids and fatty acids)disrupts the
physiochemical integrity of the lens membrane resulting to Na+ extrusion pump failure and thus
gain in Na+ ions and water flows into cells. There would be intumescence (swelling) and loss of
lens transparency.

ENERGY UTILILIZATION OF THE LENS

All living cells require energy for survival by performing a multitude of tasks.The lens is no
exception as it requires energy for activities such as cell division, movement,growth,protein
synthesis, transport of nutrients and wastes,dehydration process and the metabolism of several
metabolites.

However, the energy requirement of the lens is absolutely low. Thus to maintain transparency so
as to sustain its optical process the lens is avscular and is surrounded by the AH and VH .These
two tissue which are fluidy in nature and also optical tissues assist in its substance.

Glucose is the major metabolite obtainable by the lens from the AH and VH which are both rich
in glucose.The glucose diffuse into the lens and is metabolized through the following pathways:

The glycolytic pathway-----Glycolysis

The krebs cycle

Pentose phosphate pathway

The sorbitol pathway.

All of these could be used. However, the glycolytic pathway is the most common. The end
products of glucose metabolism are lactic acids, CO2 and H2o.The lactic acid in the lens is
eliminated through diffusion into AH. About 80% of glucose

metabolized is by glycolytic pathway in which the end products are lactic acid and ATP.
TEARS FILM

The tear film is defined as the that surface of the eye, which remains mostly directly in contact
with environment .The precorneal tear film form a film between the inside of the lids and the
cornea when eyes are closed.it remains as a film over the cornea for between 15 and 20 seconds
after the lids are opened.

The tear film is a highly specialized and well organized moist film which covers the bulbar and
palpebral conjunctiva and cornea. The tear film is formed and maintained by an elaborate system
that comprises the lacrimal apparatus, which consist of the secretory distributive and excretory
part.

The secretory parts includes the lacrimal gland, the accessory lacrimal gland tissue, the
sebaceous glands of the eyelids, globlet cells and other mucin secreting elements of the
conjunctiva.

The distributive parts involved eyelid movement

The excretory part or elimination part of these secretion is based on the movement of tears
across the eye aided by the act of blinking sending the tears through a drainage system consisting
of lacrimal punta , canaliculi, the sac and the nasolacrimal duct.

FUNCTION OF TEAR FILM.

1. It is critically important for protecting the eye from external influences and for maintaining the
health of the underlying cornea and conjunctiva.

2. The optical stability and normal function of the eye depend on an adequate supply of fluid
covering its surface thus maintaining an optically uniform corneal surface.

3. A mechanical function is by flushing off cellular debris, foreign matter from the cornea and
conjunctiva sac.

4. Lubrication of the corneal surface and also the bulbar and palpebral conjunctival surface

5. Nutritional function by feeding the cornea which is avascular with nutrients

6. Tears also possess immunochemical properties

Protect the eye from damage by foreign bodies, irradiation, and dust. Etc.

READ UP ANATOMY OF THE LACRIMAL GLAND

COMPOSITION OF TEARS

Tears contain 98.2% water and 1.8% solid. The high percentage of water in tears is a natural
consequence of the need for lubrication of the conjunctival and corneal surface. The continuous
evaporation of water between blinks may influence the water concentration of the tear film.

Physical properties of tears

 A film is a thin layer that can stand vertically without gravitational flow.
 Tear film are clear and transparent giving it the optical qualities
 Tear film is a composed of 3 layers-superficial lipid layers, middle Aqueous layer,
posterior .mucin layer all arranged according to their densities.
 The tear film is divided in three part.—portion covering the palperbral conjunctival,
portion covering the bulbar conjunctiva. precorneal tear film covering the cornea
 As an optical apparatus the refractive index of tears is 1.357
 The viscosity of tears is almost that of water
CHEMICAL PROPERTIES OF TEARS

PH -the pH of unstimulated tears is about 7.4 which approximates that of blood

plasma .however, a wide variation is found in normal individuals ranging between 5.0-8.35 but
common range is between 7.3 and 7.7 A more acidic pH of about 7.25 is found following
prolonged lid closure possibly due to carbon dioxide produced by the cornea and trapped in tear
pool inside the eyelids. Tear pH is characteristic of the individual and normal buffering activity
maintains the pH as relatively constant level especially diurnally.

OSMOTIC PRESSURE- the osmotic pressure in tears results by the presence of electrolytes
which is 305mosm/kg. Equivalent to 0.05%Nacl. At day time, the osmotic pressure fluctuates
between 0.9 to 1.02% Nacl equivalents. During prolong lid closure the osmotic pressure could
decrease up to 285mosm/kg equivalent to 0.89%Nacl increase in aqueous component results to
hyper tonicity of the tear and when decreased, results to hypo tonicity.
Hypo tonicity increased when lid is open and there is evaporation while hyper tonicity result in
lid closure .Osmotic changes assert in regulation of tear flow-Reflex stimulation of tears during
early adaptation to contact lenses result to decrease in both electrolyte and protein content
leading to hypertonocity which accounts for corneal edema experienced at this stage.

CHEMICAL COMPOSITION OF TEARS

Immunoelectrophoretic studies have shown that tears contain lipids protein, enzymes, several
metabolites, electrolyte, and hydrogen ions etc

PROTEIN CONTENT

Perhaps the most striking features in the chemical analysis of tears is the relatively high
concentration of protein. The presence of protein gives the tears much lower tensions and
enables it to wet the epithelial surface more perfectly. Secondly, it enables them to produce a
stable foam on account of absorption of the protein molecules at the interphases between air, the
tears and the cornea. This is of considerable significant as the transparency of the cornea partly
depends on it. Moreover, the optical properties of the eye are greatly improved since microscopic
irregularities in the corneal epithelium are abolished by the presence of proteins thereby
abolishing appearance of bumps, thus producing a perfectly smooth and polished optical
surface for the eye. About 60 varieties of tear protein have been isolated. This clearly
distinguish the tear protein with that of blood plasma.

Group A tear proteins are similar to serum proteins representing just 15%of all tears proteins.
These include Albumin, IgG,transferrin ᾳ-L antichymotrypsin and B-2 microglobulin.

GROUP B tear protein- these are specific proteins synthesized by tear glands.The rapid
migration protein (RMP),lysozymes,lactoferrin and IgA.These are produced as external
secretions.

Tears albumin constitutes about 60% total protein in tears as also applicable to the plasma.
Electrophoresis of albumin shows several peaks of migration which indicates that this is
produced by the lacrimal gland. Generally, the total protein content depends strongly on the
method of collection of tears. Unstimulated tears show levels of about 20mg/ml while stimulated
tear show lower value of 1-7mg/ml.
LYSOZYME

Fleming in 1922 discovered that tears contain a substance that dissolved or lyze many airborne
saprophytes rapidly and completely and so named it lysozyme. This enzyme is also found in
nasal mucosa (mucus), sputum and various tissue extracts, vegetable juices and egg white.
Normal tear concentration of lysozyme is much higher than other body fluid. The Normal tear
lysozyme level us 1-2mg/ml and the enzymatic activity is optimal at PH of 5.2 and decrease
above or below this level. Lysozyme is a long chain, high molecular weight protolytic enzyme
produced by lysozyme a known cellular ultrastructure. Human tears lysozyme (HTL) levels have
been shown to be greatly decreased in tears of patient suffering from sjogren’s syndrome and
ocular toxicity from long term use of practolol therapy. HTL levels is lowered in Herpes simplex
infection and HIV infection, malnutrition in children.

LIPIDS

Lipids are present in small amount in tears as they are found in the very thin superficial lipid
layer of the tear film. Major types of lipids includes waxy esters, hydrocarbons, triglycerides,
cholesterol esters and as much lesser amount of digycerides, monoglyerides ,free fatty acid, free
cholesterol and phospholipid

CHOLOESTEROL

The amount of cholesterol in tears is the same as that found in blood .the concentration of
cholesterol in tears is same as that of blood. Cholesterol like other lipids, is transported by ᾳ and
ß lipoproteins.

METABOLITES

Glucose

In normal tears, there is 0.2mmol/l in tears fluids though it has not been proven it has always
been believed that tear film glucose feed the cornea. The glucose from tears come from the
goblet cell of the conjunctiva. There has been corresponding rise in tears glucose level with
elevation of plasma glucose level that is above 100mg%.this implies that In diabetic there would
be change in glucose level.

UREA

Because of the unrestricted passage of urea through the blood tear barrier in the lacrimal gland,
there is a balance between urea concentration in tears and plasma. However, urea concentrations
in tears decrease with increasing secretion rate.

ELECTROLYTES AND HYDROGEN ION

Cations which are positively charged electrolytes are much more predominant in tears than
anions. Cations include sodium and potassium while anions are chloride and bicarbonate.

SODIUM the sodium concentration of tears is 120-170mmol and is equal t that of plasma
suggesting a passive secretion into the tears. There is an active secretion of sodium into the tears.
The physiology behind this is that while there is high concentration of sodium in the ocular
fluids, the cornea has rather a higher essential role in the osmotic regulation of the intra and
extracellular spaces and in general, changes in sodium level are the reverse of changes in
potassium.

CALCIUM though independent of tear pathophysical, its level in tears is low .However, when
high, it is used as a marker for assessment of disease such as cystic fibrosis.

Chloride

chloride concentration is slightly higher in tears than in plasma .it is important for osmotic
regulation of all tissues.

READ UP LAYERS OF THE TEARS FILM

THE AQUEOUS HUMOUR

The aqueous humor is a clear slightly alkaline watery fluid occupying both the anterior and
posterior chambers of the eye. It is a dynamic and continuously secreted and passed over the lens
through the pupil into the anterior chamber and subsequently drained into the vascular system-
specifically the anterior ciliary vein. This fluid penetrate the gel like vitreous humor and is
poorly identical with that found in the intercellular spaces of the various ocular coats. The AH
has a density slightly higher than water and the volume of the anterior chamber is about 0.255ml
while that of the posterior chamber is about 0.06ml.due to low metabolic activities of adjoining
tissues, whatever circulation of fluid exist is sufficient to meet the metabolic demands of these
structures which depends on it for their nutrition

Also, the lens and cornea are devoid of blood vessels implying that their nourishment must be
supplied largely by the bathing fluid and waste products must be carried away in a similar
fashion. The aqueous must therefore contain the building blocks necessary for tissues
replacement .i.e sugars, amino acid etc. moreover, it must transport oxygen and remove waste
products of metabolism, including carbon dioxide for the two chamber to the vasculature. These
functions are expected to be performed by a normal eye.

CHEMICAL COMPOSTION OF THE AH

Analytical determination of the chemical constituents of AH are quite valuable .this is because
such analyses have been shedding light on the underlying mechanisms concerned with the
formation of the fluid. The AH, is composed predominately of electrolyte and low molecular
weight compounds with some proteins and there are significant differences from plasma .they
also include steroid, sex hormones, enzymes such as carbonic anhydrase .the protein content of
aqueous is very low compared to plasma (about 1:500) these account for the difference in the
solid component of blood and aqueous humor. The solid component of AH contains 9.5g/100ml
while blood has 10.8g/100ml.hence the AH is much more diluted than blood serum.it is
important to note that during inflammation the protein content increases to 10g/100ml due to a
breakdown of the blood aqueous barrier .the blood aqueous barrier is a tight junction between the
epithelial cell of the ciliary body and non- fenestrated iris vessels. Still on inflammation the IgG
is present in AH in a concentration of 3mg/100ml. IgA , IgM. IgD are not present but during
inflammation of the eye, their presence is noticed.

Glucose conc in AH is 80%that of plasma. Glucose gain entrance into AH through simple
diffusion.
Ascorbic acid content is higher than in blood .the increases in ascorbate concentration is usually
fast and is secreted by active transport usually by a specific transport mechanism that can be
saturated.

AQUEOUS HUMOUR PRODUCTION

Production or formation of AH occurs in the secretory ciliary epithelium by 3 basic mechanisms

Diffusion ---this occurs in the side of greater concentration to the side of lower concentration.
This is made possible for lipid soluble substances that by this process penetrate all membrane
readily.(diffusion of water and ions from the fenestrated vessels of the ciliary body )

ACTIVE SECERETION----active secretion or active pumping of substances into the anterior

chamber is the major means of AH production. Such secretion is the result of an active metabolic
process and is independent of the level of IOP. The active transport of certain solutes by the
ciliary epithelium is the most important in aqueous humor formation this is made possible
through the Na +/K+ATPase transport system. (Followed by the active secretion of the Na /K
ATPase)

ULTRAFILITRATION

The rate of ultra-filtration is influenced by the level of blood pressure in the ciliary capillaries,
the plasma oncotic pressure and the level of IOP. The process of ultrafiltration of fluid from the
plasma into the posterior chamber is responsible for as much as 75% of AH formation
(ultrafiltration of ions and water and eventually leads to the formation of aqueous humor).

VITREOUS HUMOUR

The vitreous body is a transparent, jelly like substance filling the posterior cavity of the eye.it is
relatively unstable and truly fragile.it is a clear mass, physically a hydrogel colorless gelatinous
mass. It has a consistency that is firmer than egg –white. In man and higher primates, the
vitreous is by far the largest part of the globe.it is bounded by the retina, ciliary body and the
posterior capsule of the lens.

PHYSICAL PROPERTIES OF VITREOUS

The vitreous fluid is an interesting mixture of fluid and gel that varies between species as well
with the age of the species.in humans, it begins as 89% gel and 20% fluid and gradually changes
to 40% gel and 60% fluid. The peculiar characteristics of the vitreous are due to their inclusion
of type ii and other collagen as well as proteoglycans. The change in proportion of the gel and
fluid with age is considered to be the result of the breakdown of type ii collagen. Unfortunately
this breakdown can result is destabilization of the retinal surface and lead to retinal detachment.
Also the vitreous possess some RESILENCE (ability to reform its original shape after
deformation and flow. Also VISOELASTICITY (ability to cushion the eye from shock by
absorbing external forces hinged upon it.)

CHEMICAL PROPERTIES OF VITEROUS

The ascorbate level of the vitreous is very high. The amount of protein are high due to the
presence of collagen ii and hyaluronic acid which contribute to the gelatinous nature of the
vitreous. Sodium and glucose level are low due to the difficulty in transportation and also due to
the high need of the retina for glucose for metabolic activity.

Collagen is the major structural protein of virtous.it is predominately high (90%).

FUNCTION OF THE VITREOUS

 -oxygenation of posteriorly located tissue of the eye such as retina.

 -serves as the metabolic repository of the retina due to the presence of glucose and other
metabolites.
 -movement of nutrient, waste solutes, solvents due to the process called transvitreal
movement which includes diffusion, active transport. etc.
 Protect the retina by absorbing dangerous radiations
 Gives the eye its oval shape
 Allows light to be guarded to the retina.

FACTORS NECESSITATING THE OPTICAL ROLE OF THE VITREOUS

 1. Media transparency—it allows the transmission of light to the retina. This is made
possible from the structural orientation of the collagen hence 90% of light rays are
transmitted.
2. Prevention of cell invasion—the presence of HA inhibits cellular migration and
proliferation. These minimizes light scattering ,maintain clarity and sustain the
gelatinous viscosity
3. PREVENTION OF NEOVASCULARIZATION AND HEMORRHAGE----
through the action of the collagen it has been proven that the vitreous has ability to
control neovascularization and heamorrhage by shortening the partial thromboplastic
time. ( i.e time taken for blood to clot)-platelet aggregation.
4. ACCOMMODATION---the vitreous assist the lens in accommodation by pushing on
the lens to alter its shape

SIMILARITY BETWEEN THE AH AND VH

I. Both are transparent hence are optical tissues
II. Both are intraocular fluids of the eye
III. Both are involved in the substance of normal IOP
IV. Both are involved in the maintenance of ocular structures
V. Their refractive indices are almost the same 1.3360 for AH and 1.3349 for VH
VI. Both transmit light to the retina
VII. Both regulate the quantity of light entering the retina

Differences
1. The AH is an ocular fluid of the same viscosity and density as water ,while the vitreous is
a gel of different viscosity and density
2. AH is secreted from the blood plasma by the ciliary process of the ciliary body while VH
is produced during the embryological stage
3. Both differ in protein content
4. The concentration of metabolite and glucose is lower in VH than is with AH.
 The Retina

The innermost layer of the eye is the retina. The ten layered membrane is bathed
in vitreous humor and contains the cells necessary for transduction of photon energy. The ten
cellular layers are listed below from the layer next to the vitreous humor to the layer closest to
the choroid:

 Retinal pigmented epithelium

 Photoreceptor layer

 Outer limiting membrane

 Outer nuclear layer

 Outer plexiform layer

 Inner nuclear layer

 Inner plexiform layer

 Ganglion cell layer

  Nerve fiber layer

 Inner limiting membrane

The retina covers the visceral surface of the globe circumferentially up to

the cilioretinaljunction (i.e. the ora serrata; the serrated border where the retina meets the ciliary
body). After passing through the optic components of the eye (i.e. cornea, lens, and humors),
light rays penetrate all the layers of the retina to reach the photoreceptor layer. Activation of the
photoreceptors then initiates the transduction cascade.

Also recall that the retina can be divided into four quadrants such that the parts closest to the
nose are referred to as the upper and lower nasal retina, and the parts closer to the temporal side
of the head are the upper and lower temporal retina. The nasal retina of the left eye and the
temporal retina of the right eye receives visual input from the left visual field. Similarly, the
upper parts of the retina receive visual stimuli from the inferior visual field, while the lower part
of the retina is stimulated by input from the upper visual field. Reversal of this logic is
applicable.

Retinal Pigmented Epithelium

The retinal pigmented epithelium is the most superficial (i.e. outermost) layer of the retina. It is
made up of simple cuboidal to low columnar epithelium that is affixed to Bruch’s membrane
(innermost layer of the choroid). The layer is characterized by numerous mitochondria-rich
invaginations into the basement membrane, multiple gap junctions, and other junctional
complexes. The apices of the cells are richly populated with melanin granules, as well as
secondary lysosomes, peroxisomes, and phagocytic vacuoles. There are also numerous smooth
endoplasmic reticula that specifically facilitate vitamin A isomerization.

The retinal pigmented epithelium is instrumental in establishing and maintaining the blood-
retinal barrier. It controls the ion exchange between the vascular choroid and the photoreceptors
layer of the retina. The cells of this epithelium also have phagocyticproperties; therefore, they are
able to clear the cellular debris generated by the photoreceptors. The pigmented layer supplies
the photoreceptors with additional adenosine triphosphate (ATP), immunomodulatory, and
polypeptide growth factor molecules to carry out the transduction process.

During photopic conditions (when the light intensity is high), the villous processes of the
pigmented epithelium elongate into the photoreceptor layer of the retina. The processes also
retract under scotopic (low-intensity light) conditions. Therefore, it is able to absorb scattered
light as it passes through the layers of the retina and protect the photoreceptors from excess light
exposure. Lastly, the pigmented layer produces antioxidants that help neutralize the free radicals
generated in the transduction process.

Photoreceptor Layer

Humans have two types of photoreceptors that are named according to the morphology of the
cell bodies. The rods are cylindrical cells that operate best in low-intensity light; while cones are
(surprise!) conical cells that function best in high-intensity light and facilitate color perception.
The rods are more often than not, longer and more slender than the cones. However, the converse
is true when comparing the photoreceptors toward the peripheral aspect of the retina. The
distribution of the photoreceptors throughout the retina is such that the rods are widely scattered
throughout the entire retina except at the fovea (minute central depression within the macula;
about 4 mm lateral to the optic disc). The fovea is solely occupied by cones (which are relatively
scarce throughout the rest of the retina). It should be noted; however, that neither rods nor cones
can be found in the optic disc.

The photoreceptors are the cells responsible for converting energy from photons to electrical
energy that can be conducted by the nerve fibers. While the actual cell bodies of the
photoreceptors are within the outer nuclear layer the peripheral connection between the
photoreceptors and the retinal pigmented epithelium (i.e. the outer segment, primary cilium, and
inner segment of each photoreceptor) form the photoreceptor layers. The axons of the rods and
cones are stimulated by energy from the photons as they reflect off the retinal pigmented layer.

Outer Nuclear Layer

Each photoreceptor has four major components. There is an outer segment, inner segment,
nucleus, and a synaptic body (spherule). The outer segment is highly folded and contains the
photosensitive chemicals necessary for initiating the visual impulse. The membrane of the outer
segment is also highly folded into discs to increase the surface area of the cell. It also contains
photochemicals (rhodopsin in rods and color pigments in cones) which are proteins that are
conjugated to the transmembrane proteins of the outer segment discs.

The inner segment is the cytoplasmic region of the cell. It houses the cell cytoplasm and other
cellular organelles integral for cell function. The mitochondria are likely the most abundant and
most important organelles in these cells, as a large amount of energy is needed to facilitate the
photoreaction. The outer and inner segments communicate via a slender stalk known as
the cilium (filled with microtubules that facilitate the signal transduction).

The nucleus is separated from the rest of the cell body by the outer limiting membrane.
Therefore, a large portion of the photoreceptor (outer and inner segments) is found in the
photoreceptor layer, while the nucleus is located in the outer nuclear layer.

The synaptic body is the innermost part of the photoreceptor. It communicates with the second
order neurons (i.e. horizontal and bipolar cells) of the inner cell nuclear layer in the outer
plexiform layer. The differences between the rods and cones are found in the table below.

Rods

Rods are ubiquitously distributed throughout the periphery of the retina. Unlike cones,
the cylindrical stacks of membrane discs are encased within a membrane. The outer segment of
the rods contains high resting levels of cyclic guanosine monophosphate (cGMP) as well as
inactive rhodopsin molecules. The inactive rhodopsin moieties are bound to the membranous
discs of the outer segment, while the cGMP molecules are continuously released in the absence
of light. Elevated cGMP levels promote an influx of sodium (Na+) ions into the outer segment;
giving rise to an elevated resting membrane potential.

Rhodopsin undergoes molecular rearrangement with exposure to light; and together with other
photosensitive molecules in the outer membrane, they cause a fall in the quantity of cGMP. As a
result of the fall in cGMP, the sodium channels close. Consequently, there is a shift in the ionic
preference of the cells and they become hyperpolarized (unlike other afferent neurons that
become depolarized following a stimulus).

Instead of initiating an action potential to spread the effect of the stimulus, the hyperpolarization
gradually moves across the cell membrane. Once at the synaptic bulb of the rods, the
hyperpolarization gradually reduces the amount of glutamate (a neurotransmitter) being
produced at the synapse (rod spherules). This can have a depolarizing or hyperpolarizing effect
depending on the cells at the other end of the synapse. Although it is able to complete a relatively
large number of photoisomerization, rhodopsin molecules eventually require replacement. Since
they are bound to the discs of the cell membrane, the distal fragment of the rod undergoes rod
shedding every ten days, and proximally the discs are renewed.

Cones

In contrast to the rods, the cones are not enclosed within a membrane and are in constant
communication with the extracellular space. Furthermore, the stacks of discs progressively get
smaller and further away from the inner segment. Therefore, the cones have a characteristic
conical shape. There are three types of cones, each responsible for detecting light within a
particular spectrum:

 The long wavelength cones (L – cones) are sensitive mostly to light within the red
spectrum.

 The medium wavelength cones (M – cones) are highly responsive to light in the green
spectrum.

 The short wavelength cones (S – cones) primarily detects light in the blue spectrum.

Therefore, the cones are responsible for color vision; the perception of which will be mitigated
by any particular combination of stimuli generated from the three cones. The photopigment of
the cones is similar to those found in the rods, with the exception that there are three different
types (one for each type of cone). Those differences aside, they are also found in the outer
segment of the cones, and also undergo light-induced conformational changes. The change in
molecular structure triggers a similar hyperpolarization reaction that gradually spreads over the
cell membrane toward the synaptic bulb of the cone (cone pedicles). The decline in glutamate
release and resultant depolarizing or hyperpolarizing phenomenon also occurs.

Inner Nuclear Layer

There are several accessory cells within the retina that form regulatory connections with the
photoreceptors. Their cell bodies are found in the inner nuclear layer of the retina (between the
inner and outer plexiform layers). It contains the cell bodies of amacrine, horizontal, and bipolar
cells.

Bipolar Cells

The type of synaptic input that enters a bipolar cell determines whether or not it will be classified
as a cone or rod bipolar cell. These cells form the bridges between the ganglion cells and
photoreceptors of the retina. They are particularly important in detecting the edges of images
sent to the visual system.
They are further categorized into “on” or “depolarizing”, or “off” or “hyperpolarizing” bipolar
cells. The “on” pathways are activated by light, while the “off ” pathways are activated during
darkness. The logic behind “on” and “off” bipolar cells can become rather discombobulating
owing to the fact that glutamate has always been regarded as an excitatory neurotransmitter. To
avoid further confusion, just think of it this way: stopping the flow of glutamate (i.e. exposing
the photoreceptors to light) causes “on” bipolar cells to depolarize, and “off” bipolar cells to
hyperpolarize.

Amacrine Cells

The small-bodied amacrine cells do not demonstrate prominent axons, but undergo significant
arborization, with their axons extending widely. Several neurotransmitters such as gamma (γ)-
aminobutyric acid (GABA), glycine or acetylcholine (ACH), and other neuropeptides can be
found in amacrine cells. They regulate the activity of bipolar cells and increase the sensitivity of
class Y ganglion cells to moving stimuli.

Horizontal Cells

The horizontal cells are characterized by cells with axons of varying lengths (to access
photoreceptors both near and far) that travel parallel to the retinal plane and numerous dendrites.
The cell bodies are restricted to the inner nuclear layer. In response to the glutamine released
from the photoreceptors, horizontal cells secrete GABA to nearby rods and cones. This action
regulates the response of the ganglion cells, as well as sharpening the periphery of the images
transmitted to the visual system.

Ganglion Cell & Nerve Fiber Layer

The second order neurons that form the bridge between the photoreceptors and the
lateral geniculate body of the thalamus are the ganglion cells. Their cell bodies are located within
the ganglion cell layer, and their nerve fibers travel in the nerve fiber layer (adjacent to the
vitreous humor) toward the optic disc. Here, they form the optic nerve.
Ganglion cells are subdivided based on morphological and physiological features. The alpha
cells have larger cell bodies, thicker axons, and diffusely-arborizing dendrites. They are more
commonly encountered in the peripheral aspect of the retina and receive stimulation from
the rods. From a physiological perspective, they are referred to as Y cells as they have little color
sensitivity. They are also referred to as M cells owing to the fact that they synapse with the
magnocellular layers of the lateral geniculate body.

The beta cells are another category of ganglion cells that have medium sized soma and fewer
dendrites. They are more common in the central aspect of the retina and receive visual stimuli
from the cones. Therefore, they respond to color stimuli; and as such, are categorized as X cells.
They are also called P cells because they synapse on the parvocellular layers of the lateral
geniculate body.

Other ganglion cells are physiologically classified as W cells and anatomically referred to
as delta, epsilon, and gamma cells. A unique group of W cells that have little to no
communication with the photoreceptors is known as the melanopsin-containing ganglion cells. In
addition to their lack of participation in visual imaging and cell size, these cells are typified by
their connection to the pretectal (main and accessory CN III) and suprachiasmatic nuclei,
location in the ganglion layer, and sensitivity to blue light. Most significantly, these cells are
extremely sensitive to light and will propagate an action potential in response to direct exposure
to light.

Plexiform Layers

There are two separate, dense fibrous networks found between the three neuronal layers of the
retina. The outer plexiform layer is found between the outer nuclear layer and the inner nuclear
layer, while the inner plexiform layer exists between the inner nuclear layer and the ganglion cell
layer. The outer plexiform layer contains the neurons of the bipolar and horizontal cells of the
inner nuclear layer, as well as the axons of the photoreceptors. There is usually a triad of
communicating processes formed among a solitary rod spherule or cone pedicle, two laterally
positioned horizontal cell processes, and a central postsynaptic bipolar cell.
On the other hand, the inner plexiform layer contains the bridging axons that connect the cells of
the inner nuclear layer (from bipolar or amacrine cells) with those of the ganglion layer. The
amacrine cells have connections with other amacrine cells, as well as with both bipolar and
ganglion cells. Bipolar cells also communicate with corresponding ganglion cells.

Limiting Membranes

In addition to the plexiform layers, there are two other partitioning membranes found throughout
the retina. There is an array of junctional complexes that form a distinct, albeit dim, layer at the
level of the rods and cones. This is known as the outer limiting membrane (layer) which
separates the photoreceptors from the Müller cell processes. An inner limiting membrane (layer)
containing the terminal processes of the Müller cells covers the peripheral part of the vitreous
body.

The Macula

On the temporal side optic disc (roughly 4 mm laterally) and opposite to the pupil is a yellow
area known as the macula lutea. At the middle of the macula is the fovea centralis; otherwise
called the fovea. Going from the periphery of the center of the macula, there is a reduction in the
thickness of the inner layers to the point that only the photoreceptor layers remain at the foveal
pit. Consequently, larger quantities of photons make it to the photoreceptors as there are fewer
intervening cells. This region is exclusively populated by cones and accounts for the majority of
the visual afferent stimuli that is transmitted to the brain.
Fovea centralis - cranial view

The Optic Nerve (CN II)

Approximately one million myelinated axons arising from the ganglion cells of the retina come
together at the optic disc to form the optic nerves (CN II). Unlike other nerves of the body that
are myelinated by Schwann cells, the fibers of the optic nerve are myelinated
by oligodendrocytes. Arising from the posterior pole of each eye, each optic nerve is about 35
mm and 55 mm in length and can be subdivided as follows:

 The intraocular part

 The intraorbital part.

 The intracanalicular part.

 The Intracranial part.

As the axons of the ganglion cells converge at the optic nerve head, there are no associated
photoreceptors in this region. As a result, any light hitting the neurons in this area will not be
perceived. Therefore, a physiological blind spot exists where no light stimuli can be appreciated.
The tubular optic nerve exits the orbit through the superior orbital fissure and meets with the
contralateral CN II above the diaphragma sella. Along this course, they become encased in layers
of dura and arachnoid mater, which are in direct communication with the same meningeal layers
of the brain. Here it forms the optic chiasm.

The Optic Chiasm & Tract (Reticulogeniculate Tract)

The optic chiasm is not only a point of union but also a point of decussation of the bilateral CN
II. Here, the nasal fibers of each eye cross the midline to join the temporal fibers of the
contralateral eye. As highlighted earlier, the nasal fibers of the right eye and the temporal fibers
of the left eye are stimulated by photons that are emitted on the right half of the visual field.
Similarly, the nasal fibers of the left and temporal fibers of the right eye receive visual stimuli
from the left half of the visual field. Following the decussation, the visual input from the right
half of the visual field will travel in the left optic tract, while the stimuli from the left half of the
visual field will pass through the right optic tract.

The optic tracts are the compact, caudolateral continuations of the retinal ganglion layer. Each
tract carries fibers that are stimulated from the contralateral visual field. As the optic tracts move
toward the ipsilateral lateral geniculate body of the thalamus, they cross over
the crus cerebri (where it joins with the cerebrum).

The Pretectal Connections of the Visual Pathway

About 1% of the retinal ganglion cells are highly responsive to light. The axons of
these melanopsin-containing ganglion cells also travel in the optic nerve. However, instead of
traveling to the lateral geniculate nucleus, they diverge to the pretectal area. This part of
the midbrain is anterior to the central aqueduct of Sylvius. The fibers then synapse on the
ipsilateral and contralateral oculomotor (CN III) and Edinger-Westphal (accessory CN III)
nuclei. These nuclei, along with other cortical influences, are responsible for several visual
reflexes including accommodation, direct and consensual light reflex, and saccadic response of
the eyes.

The Lateral Geniculate Body

Laterally and inferiorly on either side of the inferior surface of the diencephalon are two rounded
protuberances known as the lateral geniculate bodies. They house the multi-layered lateral
geniculate nuclei that are critical in processing vision. The six layers of the lateral geniculate
nucleus contain cells that are similar to those of the ganglion layer of the retina. The fibers
arriving from the ipsilateral optic tract form the ventral base of the nucleus, while the outflow
tracts that form the optic radiation also form the lateral and dorsal borders. The layers are
numerically labeled one to six, with the larger magnocellular (M) cells being restricted to layers
one and two, and the smaller parvocellular (P) cells found throughout layers three to six.

The magnocellular layer receives input from the Y cells of the ganglion layer. As a result, they
receive input from a larger visual field, respond to moving stimuli, and obtain stimuli primarily
from rods. In contrast, the parvocellular layer is stimulated by X cells. Therefore, they respond to
color stimuli of high acuity, receive input from a smaller area, and respond to stationary stimuli.
The remaining W cells terminate on the intervening thin layers of the lateral geniculate body.

Further retinotopic division of the lateral geniculate body relates to the decussation that occurred
at the chiasm. The axons arising from the nasal retina that crossed the midline eventually
terminate on layers 1, 4, and 6 of the contralateral geniculate nucleus. While the axons of the
ganglion fibers originating from the temporal retina that did not cross the midline will terminate
on layers 2, 3, and 5 of the ipsilateral geniculate nucleus.

Optic Radiation & Visual Cortex

The optic radiation (geniculocalcarine tract) represents the visual tracts that extend from
the lateral geniculate body to the primary visual cortex (Brodmann 17) on the same side.
The retinotopic distribution of the nerve fibers also continues along this path. Like the
corresponding optic tract, the ipsilateral optic radiation only carries visual input from the
contralateral visual field. The optic radiation is separated into two main tracts that correspond to
the upper and lower quadrants of the respective visual field.

Optic radiation - axial view

The second order axons corresponding to the contralateral upper quadrant originate from the
ventrolateral aspect of the lateral geniculate nucleus. They are referred to as Meyer’s loop and
take an indirect course through the white matter of the temporal lobe to access the inferior bank
of the calcarine sulcus of the lingual gyrus.

On the other hand, the second order neurons responsible for the contralateral lower quadrant
emerge from the dorsomedial part of the lateral geniculate nucleus. They have a more direct
course to the superior bank of the calcarine sulcus of the cuneus, as they pass through the
retrolenticular part of the internal capsule. The tracts corresponding to the macula and fovea are
referred to as the geniculostriate fibers and they arise from the center of the lateral geniculate
nucleus to access the caudal visual cortex

PHOTOCHEMISTRY OF VISION

The shape of an adult human eye is like a globe with a diameter of approximately2.5 cm, that
fits into the bony sockets in the skull. The globe of the eye or eyeball is a three-layered structure
which consists of sclera, choroid and retina (Clegg and Mackean 2000).

The sclera is a tough but elastic, white outer layer of connective tissue. At the front of the eye,
the sclera becomes the transparent cornea, which allows light to enter the interior of the eye and
functions as the first constituent of the light focusing system of the eye (Keeton and Mc Fadden
1983). A delicate layer of epithelial cells forms a mucous membrane, known as the conjunctiva,
which covers the outer surface of the sclera and helps to keep the eye moist (Clegg and Mackean
2000).The choroid is a layer of darkly pigmented tissue through which many blood vessels pass
and is located just inside the sclera. The choroid is important since it provides blood to other
parts of the eye and it functions as a light absorbing layer which prevents internally reflected
light from ,blurring the image. At the immediate back of the junction between the main part of
the sclera and the cornea, the choroid becomes thicker with smooth muscles embedded; this part
of the choroid is known as the ciliary body.
The front choroid forms the donut-shaped iris which gives the eye its color. The iris consists of
smooth muscle fibers arranged in circular and radial directions..

By changing size, the iris regulates the amount of light entering the pupil, the hole in the centre
of the iris. The pupil is reduced when the circular muscle fibres contract and it is dilated when
the radial muscle contract (Keeton and Mc Fadden 1983). Just inside the choroid, the retina
forms the innermost layer of the eyeball and contains the photoreceptors (Clegg and Mackean
2000).The photoreceptors are of two types, referred to as rods and cones. The rod cells are
abundant toward the periphery of the retina while the cone cells are abundant in the
central portion of the retina. The bipolar cells which are short sensory neurons synapse with the p
hotoreceptors in the retina. The bipolar cells synapse in the retina with longer neurons,
i.e.ganglion cells, whose axons form the optic nerve that runs to the visual centres of the
brain .There are several sets of synapses present in the retina, which allows the eye to modify the
information transmitted from the receptor cells to the brain (Keeton and Mc Fadden 1983).There
are no rods and cones present in the optic disc, and as such this region on the lower outside of the
retina is a blind spot, i.e. light focused on that part is not detected. The theoretical line through
the center of the lens is referred to as the optical axis (Clegg andMackean 2000).

The fovea or “yellow spot” lies on the optical axis and is the place of most acute vision. This
portion contains many cones but few rods (Vines and Rees 1972; Cleggand Mackean 2000).The
second constituent of the light-focusing system of the eye is the lens which is suspended just
behind the pupil by a suspensory ligament attached to the ciliary body (Keeton and Mc Fadden
1983). The lens and the ciliary body divide the eye into two cavities. The ciliary body constantly
produces the clear, watery aqueous humour that fills the front cavity of the eye. The back cavity,
filled with the jelly-like vitreous humour constitutes most of the volume of the eye. The aqueous
and vitreous humour function as liquid lines that helps focus light onto the retina. The lens itself
is a transparent protein disc that focuses an image on the retina (Vines and Rees 1972).The eye is
similar to a camera. The cornea and lens, which are two constituents of the light-focusing
system, form an inverted image on the retina. The iris regulates the opening of the lens while the
eyelids prevent light from entering and also prevents any possible damage to the surface of the
cornea. The ciliary muscle controls the lens so that objects from different distances may be
brought sharply into focus. The focusing of light onto the retina can be accomplished by this
mechanism and also by the curvature of the cornea (Vines andRees 1972). The cornea has
a refractive index of 1.38; the lens is 1.42 whereas the refractive index of both humours is 1.33.
The largest difference in refractive index occurs between the air and cornea and therefore it is
essential for image formation. The delicate and accurate control is achieved by the lens which
acts as a fine adjustment (Vines and Rees 1972)

Photoreceptors: Rods and Cones

The retina contains millions of photoreceptor cells. These are referred to as rods and cones and
the names of these cells come from their individual shapes. The human retina contains
approximately 125 million rods and 6 million cones. The rod cells are abundant toward
the periphery of the retina while the cone cells are abundant in the central portion of the
retina(Keeton and Mc Fadden 1983). Each rod cell or cone cell has an outer segment with a stack
of folded membranes or discs, in which visual pigments are embedded (Kimball 1983).The
visual pigment in the rods is built into the membranes of the flattened vesicles in the outer
segment and is referred to as rhodopsin (Keeton and Mc Fadden 1983). In the cones, the visual
pigment is known as iodopsin (Clegg and Mackean 2000). They are thought to be three types of
cone cells which contain different forms of iodopsin and as such red green and the blue

VISUAL PHOTOTRANSDUCTION is the sensory transduction of the visual system. It is a

process by which light is converted into electrical signals in the rod cells, cone
cells and photosensitive ganglion cells of the retina of the eye.

The visual cycle is the biological conversion of a photon into an electrical signal in the retina.
This process occurs via G-protein coupled receptors called opsins which contain
the chromophore 11-cis retinal. 11-cis retinal is covalently linked to the opsin receptor via Schiff
base forming retinylidene protein. When struck by a photon, 11-cis retinal undergoes photo
isomerization to all-trans retinal which changes the conformation of the opsin GPCR leading
to signal transduction cascades which causes closure of cyclic GMP-gated cation channel, and
hyperpolarization of the photoreceptor cell.
Following isomerization and release from the opsin protein, all-trans retinal is reduced to all-
trans retinol and travels back to the retinal pigment epithelium to be "recharged". It is
first esterified by lecithin retinol acyltransferase (LRAT) and then converted to 11-cis retinol by
the isomerohydrolase RPE65. The isomerase activity of RPE65 has been shown; it is still
uncertain whether it also acts as hydrolase. Finally, it is oxidized to11-cis retinal before traveling
back to the rod outer segment where it is again conjugated to an opsin to form new, functional
visual pigment (rhodopsin).

Photoreceptors

The photoreceptor cells involved in vision are the rods and cones. These cells contain
a chromophore (11-cis retinal, the aldehyde of Vitamin A1 and light-absorbing portion) bound to
cell membrane protein, opsin. Rods deal with low light level and do not mediate color vision.
Cones, on the other hand, can code the color of an image through comparison of the outputs of
the three different types of cones. Each cone type responds best to certain wavelengths, or colors,
of light because each type has a slightly different opsin. The three types of cones are L-cones, M-
cones and S-cones that respond optimally to long wavelengths (reddish color), medium
wavelengths (greenish color), and short wavelengths (bluish color) respectively. Humans have a
trichromatic visual system consisting of three unique systems, rods, mid and long-wavelength
sensitive (red and green) cones and short wavelength sensitive (blue) cones

Process
The absorption of light leads to an isomeric change in the retinal molecule.

To understand the photoreceptor's behaviour to light intensities, it is necessary to understand the

roles of different currents.

There is an ongoing outward potassium current through non gated K+-selective channels. This
outward current tends to hyperpolarize the photoreceptor at around -70 mV (the equilibrium
potential for K+).

There is also an inward sodium current carried by cGMP-gated sodium channels. This so-called
'dark current' depolarizes the cell to around -40 mV. Note that this is significantly more
depolarized than most other neurons.

A high density of Na+-K+ pumps enables the photoreceptor to maintain a steady intracellular
concentration of Na+ and K+.

In the dark

Photoreceptor cells are unusual cells in that they depolarize in response to absence of stimuli or
scotopic conditions (darkness). In photopic conditions (light), photoreceptors hyperpolarize to a
potential of -60mV. It is this 'switching off' that activates the next cell and sends an excitatory
signal down the neural pathway.

In the dark, cGMP levels are high and keep cGMP-gated sodium channels open allowing a
steady inward current, called the dark current. This dark current keeps the cell depolarized at
about -40 mV.

The depolarization of the cell membrane in scotopic conditions opens voltage-gated calcium
channels. An increased intracellular concentration of Ca2+ causes vesicles containing
neurotransmitters, to merge with the cell membrane, therefore releasing the neurotransmitter into
the synaptic cleft, an area between the end of one cell and the beginning of another neuron. The
neurotransmitter released is glutamate, a neurotransmitter whose receptors are often excitatory.

In the cone pathway glutamate:

 Hyperpolarizes on-center bipolar cells. Glutamate that is released from the

photoreceptors in the dark binds to metabotropic glutamate receptors (mGluR6), which,
 through a G-protein coupling mechanism, causes non-specific cation channels in the cells
to close, thus hyperpolarizing the bipolar cell.

 Depolarizes off-center bipolar cells. Binding of glutamate to ionotropic glutamate

receptors results in an inward cation current that depolarizes the bipolar cell.

In the light

Representation of molecular steps in photoactivation (modified from Leskov et al., 2000).

Depicted is an outer membrane disk in a rod. Step 1: Incident photon (hν) is absorbed and
activates a rhodopsin by conformational change in the disk membrane to R*. Step 2: Next, R*
makes repeated contacts with transducin molecules, catalyzing its activation to G* by the release
of bound GDP in exchange for cytoplasmic GTP, which expels its β and γ subunits. Step 3: G*
binds inhibitory γ subunits of the phosphodiesterase (PDE) activating its α and β subunits. Step
4: Activated PDE hydrolyzes cGMP. Step 5: Guanylyl cyclase (GC) synthesizes cGMP, the
second messenger in the phototransduction cascade. Reduced levels of cytosolic cGMP cause
cyclic nucleotide gated channels to close preventing further influx of Na+ and Ca2+.

1. A light photon interacts with the retinal in a photoreceptor cell. The retinal undergo
isomerization, changing from the 11-cis to all-trans configuration

2. Retinal no longer fits into the opsin binding site.

3. Opsin therefore undergoes a conformational change to metarhodopsin II.

4. Metarhodopsin II is unstable and splits, yielding opsin and all-trans retinal.

 5. The opsin activates the regulatory protein transducin. This causes transducin to dissociate
from its bound GDP, and bind GTP, then the alpha subunit of transducin dissociates from
the beta and gamma subunits, with the GTP still bound to the alpha subunit.

6. The alpha subunit-GTP complex activates phosphodiesterase or PDE.

7. PDE breaks down cGMP to 5'-GMP. This lowers the concentration of cGMP and
therefore the sodium channels close.

8. Closure of the sodium channels causes hyperpolarization of the cell due to the ongoing
efflux of potassium ions.

9. Hyperpolarization of the cell causes voltage-gated calcium channels to close.

10. As the calcium level in the photoreceptor cell drops, the amount of the neurotransmitter
glutamate that is released by the cell also drops. This is because calcium is required for
the glutamate-containing vesicles to fuse with cell membrane and release their contents.

11. A decrease in the amount of glutamate released by the photoreceptors causes

depolarization of On center bipolar cells (rod and cone On bipolar cells) and
hyperpolarization of cone off-center bipolar cells.

Deactivation of the phototransduction cascade

GTPase Accelerating Protein (GAP) interacts with the alpha subunit of transducin, and causes it
to hydrolyse its bound GTP to GDP, and thus halts the action of phosphodiesterase, stopping the
transformation of cGMP to GMP.

Guanylate Cyclase Activating Protein (GCAP) is a calcium binding protein, and as the calcium
levels in the cell have decreased, GCAP dissociates from its bound calcium ions, and interacts
with Guanylate Cyclase, activating it. Guanylate Cyclase then proceeds to transform GTP to
cGMP, replenishing the cell's cGMP levels and thus reopening the sodium channels that were
closed during phototransduction.

Finally, Metarhodopsin II is deactivated. Recoverin, another calcium binding protein, is normally

bound to Rhodopsin Kinase when calcium is present. When the calcium levels fall during
phototransduction, the calcium dissociates from recoverin, and rhodopsin kinase is released,
when it proceeds to phosphorylate metarhodopsin II, which decreases its affinity for transducin.
Finally, arrestin, another protein, binds the phosphorylated metarhodopsin II, completely
deactivating it. Thus, finally, phototransduction is deactivated, and the dark current and
glutamate release is restored. It is this pathway, where Metarhodopsin II is phosphorylated and
bound to arrestin and thus deactivated, which is thought to be respons`ible for the S2 component
of dark adaptation. The S2 component represents a linear section of the dark adaptation function
present at the beginning of dark adaptation for all bleaching intensities.

All-trans retinal is transported to the pigment epithelial cells to be reduced to all-trans retinol,
the precursor to 11-cis retinal. This is then transported back to the rods. All-transretinal cannot
be synthesised by humans and must be supplied by vitamin A in the diet. Deficiency of all-
trans retinal can lead to night blindness. This is part of the bleach and recycle process of
retinoids in the photoreceptors and retinal pigment epithelium.