BIOCHEMISTRY OF VISION

Redario C. Laygo, M.D.

Department Of Biochemistry, Molecular
Biology & Nutrition
 Five Layers of the Cornea
Layers Thickness (in µm) Composition
Stratified Squamous
Epithelium (Ep) 50
Epithelium
Compact layer of
Bowman's
8-14 unorganised collagen
Membrane (BM)
fibres
Orderly arrangement
Stroma (SP) 500 of collagen lamellae
with keratocytes
Descemet's Consists of basement
10-12
Membrane (DM) membrane materials
single layer of simple
Endothelium (En) 5
squamous epithelium
 Stroma and Endothelium
n dense tissue of the stroma accounts for 90% of the
total corneal thickness
n heterodimeric complexes of type I and type V
collagen fibres form the parallel arrangement of
lamellae formed to maintain transparency (Fini and
Stramer, 2005).
n collagen fibres are held in a uniform spacing pattern
by proteoglycans (Lumican and Decorin)
n Keratocytes (fibroblasts) are located between the
lamellae (Hay et al., 1979)
 Stroma and Endothelium
n Keratocytes (fibroblasts) are located between the
lamellae (Hay et al., 1979)
- sparsely located linked to one another via
dendritic processes (Muller et al., 1995)
- produce crystalline proteins to maintain corneal
transparency (Jester et al., 1999).
 Descemet's membrane
n - rests on the innermost surface of the cornea
- acts as a basement membrane for the inner
endothelial cell monolayer
- cells:
> transport nutrients from the aqueous
humour to the stroma
> concurrently pump out excess water
preventing corneal edema (swelling) by
maintaining optimal hydration
 Characteristics of Corneal Collagen
Fibers Accounting for the Corneal
Transparency
n the fibers are highly uniform in diameter (25-35
nm)
n distance between two corneal fibers is also
highly uniform (41.5nm)
Lumican &
Decorin
 Corneal Transparency
n mainly dependent on:
- arrangement of these collagen fibers in stroma
> mean diameter of individual collagen fiber
> mean distance between collagen fibers are
almost the same
> both of them are less than half of the
wavelength of visible light (400-700nm)
- destructive interference - scattering of an
incident ray of light by a collagen fiber is
cancelled by interference from other scattered
rays of light
 CORNEA
Energy Source Of Cornea

GLUCOSE – major metabolic fuel

30% metabolized by EMP

65% metabolized by HMP

 Aqueous Humour
n Composition:
n Water: 99%
n Ions: HCO3-, buffers metabolic acids; Cl-, preserves
electric neutrality; Na+; K+; Ca2+; PO42-
n Proteins: albumin, β-globulins. Very low density due
to filtration
n Ascorbate: anti-oxidative, protects against UV.
n Glucose
n Lactate: produced by metabolism of anaerobic
structures of the eye
n Amino acids: transported by cilary epithelial cells
 Aqueous Humour
n Functions:
n Maintains the intraocular pressure and inflates the
globe of the eye
n Provides nutrition for the avascular ocular tissues:
posterior cornea, trabecular meshwork, lens, and
anterior vitreous
n Carries away waste products from metabolism of the
above avascular ocular tissues
 Aqueous Humour
n Functions:
n May serve to transport ascorbate in the anterior
segment to act as an anti-oxidant agent
n Presence of immunoglobulins indicate a role in
immune response to defend against pathogens
n Its main function is to provide diopteric power to the
cornea
PROTECTIVE MECHANISM OF CORNEA

n Active O2 species

n Glutathione reductase

n Glucose 6 PO4 dehydrogenase

GSSG + NADPH + H+ 2GSH + NADP+

n Aldehyde dehydrogenase (ALDH3A1)

 LENS
n Composition - water
- proteins

n Blood Supply – none

n Metabolism - active thru aqueous humor

n State - clear crystalline state

Changes in Focal Distance with Age

Age (years) Focal distance (in.)

10 2.8
20 4.4
35 9.8
45 26.2
70 240.0
 PROTEINS OF LENS
n Crystallins – α, β, γ

n Albuminoids

n Enzymes

n Membrane proteins
 NATIVE UNAGGREGATED STATE OF
THE LENS
INSULTS: PROTECTIVE
MECHANISMS:

Redox state Glutathione

UV irradiation reductase

Osmolarity Na+K+ ATPase

[Metabolites] Protein synthesis

Ways GSH or Its Depletion Affect Lens
Opacity
n mechanisms of cataract prevention by GSH:
(1) maintaining sulfhydryl (SH) groups on
proteins in their reduced form preventing
disulfide cross-linkage
(2) protecting SH groups on proteins important
for active transport and membrane
permeability
(3) preventing oxidative damage from
hydrogen peroxide (H2O2)
ENERGY SOURCE OF THE LENS

GLUCOSE

85% metabolized by EMP

(anaerobic)

10% metabolized by HMP

3% metabolized by TCA
 CATARACT

n Disease of the lens

n Opacities
n Cause: - loss of normal osmolarity
- lens protein solubility change
n Kinds: - senile
- diabetic
n Treatment – lens replacement
 SENILE CATARACT

n Cause - architectural arrangement of lens

proteins altered

- breakdown of proteins molecules:

> starting at C-terminal end
> deamidation
> aspartyl residues racemization
 DIABETIC CATARACT

n Loss of normal osmolarity of lens proteins

n Cause - increase activity of polyol pathway

n Enzymes involved:
aldose reductase
polyol dehydrogenase
 RETINA

Energy Source:

GLUCOSE
Metabolized thru EMP
(anaerobically)
 RETINA

n Visual Cells:
Rods & Cones
n Vascular
fovea centralis – no blood vessels
n Mitochondria
outer segments of rods & cones – no
mitochondria
n Enzyme - active lactate dehydrogenase
 VISUAL PIGMENTS

n Rods cells
rhodopsin
(11 cis retinal)

n Cone
red (porphyropsin) (dehydroretinal)
green (iodopsin),
blue (cyanopsin) pigments
 RHODOPSIN

n 40kDa

n Composition - protein opsin

- 11-cis retinal

n Seven transmembrane α helices

n Lysine 296 of the seventh helix

 11-cis RETINAL

n Sources:
Vitamin A

β carotene

Retinlyl esters
 TRANSDUCIN

n Classical type of G-protein

n Trimeric: α, β, γ subunits

n Two forms: inactive – αGDPβγ complex

active – αGTPcomplex
 PHOSPHODIESTERASE (PDE)

n Heterotetrameric protein

n PDE of rod cells – α,β,γ2 subunits

of cone cells - α2,γ2 subunits
 Deactivation of the phototransduction
cascade
n GTPase Activating Protein (GAP)
n Guanylate Cyclase Activating Protein (GCAP)
n Deactivation of Metarhodopsin II thru:
recoverin
arrestin
 CONES

n Color vision – trichromatic

n Three types of cells defined by visual

pigments:
cyanopsin - blue pigment 420nm
iodopsin - green pigment 535nm
porphyropsin- red pigment 565nm
 DIFFERENCES BETWEEN RODS AND
CONES
CONES RODS
1 – 3 pA 10 fA

Response time faster Response time slower

than rods – 4x than cones

Suited for rapidly Suited for low-light

changing events visual sensitivity
Differences between Rods & Cones

Rods Cones
Response kinetics slower faster
Sensitivity greater lesser
Pigment stability greater lesser
Frequency of spontaneous lower greater
isomerization
 COLOR DISCRIMINATION

Visual pigment 11-cis retinal’s (π)

protein double-bond system

Energy level
+
Absorption spectra
 COLOR VISION IS TRICHROMATIC

n Deuteranopia - absence of iodopsin

(green pigment)(medium)

n Protanopia - absence of porphyropsin

(red pigment)(long)

n Tritanopia- absence of cyanopsin

(blue pigment)(short)
 GENES OF VISUAL PIGMENTS

n Rhodopsin gene n chromosome 3

n Blue pigment gene n chromosome 7

n Red and green n X chromosome

pigment
Recharging – all trans
Retinol

http://en.wikipedia.org/wiki/Visual_cycle
(lecithin-retinol acyltransferase)

(isomerohydrolase)
 Why supplement vitamin A
n all-trans retinal is transported to the pigment
epithelial cells to be reduced to all-trans retinol, the
precursor to 11-cis retinal(recharging – all trans
retinol)
n transported back to the rods(recharging – all trans
retinol)
n all-trans retinol cannot be synthesised by humans
n deficiency of all-trans retinol can lead to night
blindness
n this is part of the bleach and recycle process of
retinoids in the photoreceptors and retinal pigment
epithelium
http://en.wikipedia.org/wiki/Visual_cycle