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Ocular Biochemistry
Tears, Cornea, Lens, Aqueous,
Vitreous, Retina and Rhodopsin
For B.Sc Optometry Students
Tear Film
Functions
• Form a smooth optical surface by filling
irregularities of corneal epithelium
• Keeps the cornea and conjuctiva moist
• Serves as a lubricant for lids
• Transfer air to the cornea
• Prevent infections
• Washes away debris and irritants
• Provides pathway to WBC in case of injury
Tear Film
Structure
1 MUCIN LAYER
• Deepest layer, 0.02-0.05 μm
• Secreted by conjunctival goblet
cells, crypts of Henle, gland of
• Stabilizes tear film, Semisolid
state, highly hydrated
• Lubricates ocular and palpebral
2 AQUEOUS LAYER
• The middle layer, Secreted by • Macromolecular muco3 LIPID LAYER
Tear Film
1 MUCIN LAYER
• Deepest layer, 0.02-0.05 μm
• Secreted by conjunctival goblet
cells, crypts of Henle, gland of
Manz and also by main Lacrinal
gland
• Made of glycoprotein, mixed
with lipids, anchored by micro
villi
• Stabilizes tear film, Semisolid
state, highly hydrated
• Lubricates ocular and palpebral
surfaces and reduces friction
• Slippery coating over foreign
bodies and protecting cornea
and conjuctiva against abrasive
effects
Tear Film
2 AQUEOUS LAYER
• The middle layer, Secreted by
lacrinal glands and accessory
glands of Krause and Wolfring
• 10 μm, It forms a uniform layer
• Lysozyme, lactoferin, tear
specific prealbumin and
immunoglobulin A
• Bicarbonate and proteins give
bufffering capacity to tears
• Macromolecular muco
glycoproteins determine the
surface tension of fluid
• Provides atmospheric O2 to
epithelium
• Washes away debris and
irritants
Tear Film
3 LIPID LAYER
• Outer most oily layer,
derived from Meibomian,
Ziess and Moll glands
• Contains lipids with low
polarity such as wax and
cholesterol esters
• Fluid layer
•Triglycerides, FFA,
phospholipids are present in
low amounts
• Prevents overflow & reduce
evaporation of tears
Tear Film
Tear Film
Chemical composition
Water
• Major component, 98.2%
• With salts dissolved Na+, K+, Cl-, HCO3-, Ca2+
Proteins
• Un-stimulated tears: 2gm/100ml
• Stimulated tears: 0.3-0.7 gm/100ml
• Group A: 15% of proteins, IgG, Albumin,Transferrin, Alpha-1
antitrypsin, Alpha1-antichymotripsin, beta-2-microglobulins.
• Group B: synthesized by tear glands, also known as rapid migrating
proteins. Lysozyme, IgA
Albumin
Tear specific protein (prealbumin)
• Acidic protein, exact function is unknown, might be stabilizing thin
tear film
Tear Film
Chemical composition
Immunoglobulins
• IgA is most prominent
• Produced by plasma cells in
conjuctiva
• defense against viral or bacterial
antigens.
• IgM and IgE are also found.
Lysozyme
• a proteolytic enzyme, net
positive charge
• Produced by acinar cells of
lacrinal gland
• work against bacterial infections
• lysis of bacterial cell wall
Glycolytic enzymes, lactate
dehydrogenase
Betalysin: antibacterial agent
Mucopoly saccharides
Glycoproteins
Amino acids
Lipids
Metabolites: glucose, lactate,
pyruvate, urea
Electrolytes: Na+, K+, Ca2+
Aqueous Humour
entire volume of fluid in the
anterior chamber is replaced 12
times a day, i.e. once every 2
hours
Aqueous Humour
Functions
• Maintenance of intraocular pressure
• They also have metabolic role
1. Cornea – takes glucose and oxygen, release
lactic acid and carbon dioxide
2. lens – use oxygen, glucose, amino acids and
potassium and release lactate, pyruvate and
sodium
• optical function : act as a diverging lens of low
power
• Clearing functions
Aqueous Humour
Biochemical composition
• Water 99.9%
• Proteins - colloid content
1. 5- 16mg/100ml
(plasma 6-7gm/100ml)
because of blood
aqueous barrier
2. IgM and IgG but NO
IgD and IgA
3. Growth factors like
FGF, TGF beta, IGF-1
4. Free amino acids
• Non colloidal contents –
dissolved solids
1. High vitamin C
2. High Pyruvate and
lactate
3. Na, K, Ca, Mg, Cl, HCO3
4. low glucose and urea
compared to plasma
Aqueous Humour
Biochemical composition
Anterior chamber
• Lesser bicarbonate
• Higher chloride
• Lesser ascorbate
• pH 7.6
Posterior Chamber
• Higher bicarbonate
• Lower chloride
• Higher ascorbate
• pH 7.57
The difference is due to
diffusional exchange across
the iris
Iris vessels are permeable to
anions and non electrolytes
Aqueous Humour: Biochemistry of formation
Aqueous Humour: Biochemistry of formation
Aqueous Humour: Biochemistry of formation
Aqueous Humour: Biochemistry of formation
Aqueous Humour: Biochemistry of formation
Active transport
• Sodium
• Chloride
• Potassium
• Ascorbic acid
• Amino acids
• Bicarbonate
Passive transport across NPE
• Water
• Chloride
• Sodium
Cornea
• Water 70%
• Enzymes necessary for
metabolism
• Acetylcholine
• Electrolytes (Na+, K+, Cl-)
Epithelium
• Collagen fibrils 70%
• soluble proteins (Ig G, A, M)
• Proteoglycans: Keratin sulphate,
• Enzymes
• Matrix metalloproteinases
(MMP-1, MMP-2, MMP-3)
• Electrolytes
Stroma
Cornea
• Water 70%
• Enzymes necessary for
metabolism
• Acetylcholine
• Electrolytes (Na+, K+, Cl-)
Epithelium
Cornea
• Lens water – Dehydrated
• Soluble Crystallins and
insoluble albumiboids
• Amino acids
• Carbohydrates
• Lipids
• Electrolytes
• Glutathione
Lens
Structure of Retina
Structure of Retina
Structure of Retina
Structure of Retina
Structure of Retina
Structure of Retina
Fovea centralis Optic Disc
Structure of Retina
Vitamin A (Retinol)
Diet
Retinol
Intestine
Blood
stream
Liver
Retinol binding
protein +
Retinol
Retinal
pigmented
epithelium
Photoreceptors
Change in Retinol
Retinol
Retinene
Rhodopsin
Retinine reductase
(oxidation)
Opsin
Inside outer segment of photoreceptors
Change in Retinol
Change in Rhodopsin
Also called as
activated
rhodopsin
Opsin
Visual Cycle
Visual Cycle
Visual Cycle : formation of nerve impulse
1. Rhodopsin changes to metarhodopsin in light
2. Metarhodopsin activates Transducin
3. Transducin activates enzyme phospodiesterase
4. Decreased intracellular cGMP
5. Closure of Na+ channel
6. Hyperpolarisation of rod cells
7. Decreased release of neurotransmitter
8. Response in bipolar cells
9. Signals to optic nerve and then to brain
Steps
• Collagen is the main content of insoluble proteins in
vitreous humor
• Hyaluronic acid is glycosaminoglycan. It adds viscosity
to the gel. It forms a water binding meshwork around
collagen fibers
• Soluble proteins has mainly glycoprotein and albumin
• Sugar
• Ascorbic acid
• Amino acids
• Electrolyte
Vitreous Humor

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Ocular biochemistry

  • 1. Ocular Biochemistry Tears, Cornea, Lens, Aqueous, Vitreous, Retina and Rhodopsin For B.Sc Optometry Students
  • 2. Tear Film Functions • Form a smooth optical surface by filling irregularities of corneal epithelium • Keeps the cornea and conjuctiva moist • Serves as a lubricant for lids • Transfer air to the cornea • Prevent infections • Washes away debris and irritants • Provides pathway to WBC in case of injury
  • 4. 1 MUCIN LAYER • Deepest layer, 0.02-0.05 μm • Secreted by conjunctival goblet cells, crypts of Henle, gland of • Stabilizes tear film, Semisolid state, highly hydrated • Lubricates ocular and palpebral 2 AQUEOUS LAYER • The middle layer, Secreted by • Macromolecular muco3 LIPID LAYER Tear Film
  • 5. 1 MUCIN LAYER • Deepest layer, 0.02-0.05 μm • Secreted by conjunctival goblet cells, crypts of Henle, gland of Manz and also by main Lacrinal gland • Made of glycoprotein, mixed with lipids, anchored by micro villi • Stabilizes tear film, Semisolid state, highly hydrated • Lubricates ocular and palpebral surfaces and reduces friction • Slippery coating over foreign bodies and protecting cornea and conjuctiva against abrasive effects Tear Film
  • 6. 2 AQUEOUS LAYER • The middle layer, Secreted by lacrinal glands and accessory glands of Krause and Wolfring • 10 μm, It forms a uniform layer • Lysozyme, lactoferin, tear specific prealbumin and immunoglobulin A • Bicarbonate and proteins give bufffering capacity to tears • Macromolecular muco glycoproteins determine the surface tension of fluid • Provides atmospheric O2 to epithelium • Washes away debris and irritants Tear Film
  • 7. 3 LIPID LAYER • Outer most oily layer, derived from Meibomian, Ziess and Moll glands • Contains lipids with low polarity such as wax and cholesterol esters • Fluid layer •Triglycerides, FFA, phospholipids are present in low amounts • Prevents overflow & reduce evaporation of tears Tear Film
  • 8. Tear Film Chemical composition Water • Major component, 98.2% • With salts dissolved Na+, K+, Cl-, HCO3-, Ca2+ Proteins • Un-stimulated tears: 2gm/100ml • Stimulated tears: 0.3-0.7 gm/100ml • Group A: 15% of proteins, IgG, Albumin,Transferrin, Alpha-1 antitrypsin, Alpha1-antichymotripsin, beta-2-microglobulins. • Group B: synthesized by tear glands, also known as rapid migrating proteins. Lysozyme, IgA Albumin Tear specific protein (prealbumin) • Acidic protein, exact function is unknown, might be stabilizing thin tear film
  • 9. Tear Film Chemical composition Immunoglobulins • IgA is most prominent • Produced by plasma cells in conjuctiva • defense against viral or bacterial antigens. • IgM and IgE are also found. Lysozyme • a proteolytic enzyme, net positive charge • Produced by acinar cells of lacrinal gland • work against bacterial infections • lysis of bacterial cell wall Glycolytic enzymes, lactate dehydrogenase Betalysin: antibacterial agent Mucopoly saccharides Glycoproteins Amino acids Lipids Metabolites: glucose, lactate, pyruvate, urea Electrolytes: Na+, K+, Ca2+
  • 10. Aqueous Humour entire volume of fluid in the anterior chamber is replaced 12 times a day, i.e. once every 2 hours
  • 11. Aqueous Humour Functions • Maintenance of intraocular pressure • They also have metabolic role 1. Cornea – takes glucose and oxygen, release lactic acid and carbon dioxide 2. lens – use oxygen, glucose, amino acids and potassium and release lactate, pyruvate and sodium • optical function : act as a diverging lens of low power • Clearing functions
  • 12. Aqueous Humour Biochemical composition • Water 99.9% • Proteins - colloid content 1. 5- 16mg/100ml (plasma 6-7gm/100ml) because of blood aqueous barrier 2. IgM and IgG but NO IgD and IgA 3. Growth factors like FGF, TGF beta, IGF-1 4. Free amino acids • Non colloidal contents – dissolved solids 1. High vitamin C 2. High Pyruvate and lactate 3. Na, K, Ca, Mg, Cl, HCO3 4. low glucose and urea compared to plasma
  • 13. Aqueous Humour Biochemical composition Anterior chamber • Lesser bicarbonate • Higher chloride • Lesser ascorbate • pH 7.6 Posterior Chamber • Higher bicarbonate • Lower chloride • Higher ascorbate • pH 7.57 The difference is due to diffusional exchange across the iris Iris vessels are permeable to anions and non electrolytes
  • 18. Aqueous Humour: Biochemistry of formation Active transport • Sodium • Chloride • Potassium • Ascorbic acid • Amino acids • Bicarbonate Passive transport across NPE • Water • Chloride • Sodium
  • 20. • Water 70% • Enzymes necessary for metabolism • Acetylcholine • Electrolytes (Na+, K+, Cl-) Epithelium • Collagen fibrils 70% • soluble proteins (Ig G, A, M) • Proteoglycans: Keratin sulphate, • Enzymes • Matrix metalloproteinases (MMP-1, MMP-2, MMP-3) • Electrolytes Stroma Cornea
  • 21. • Water 70% • Enzymes necessary for metabolism • Acetylcholine • Electrolytes (Na+, K+, Cl-) Epithelium Cornea
  • 22. • Lens water – Dehydrated • Soluble Crystallins and insoluble albumiboids • Amino acids • Carbohydrates • Lipids • Electrolytes • Glutathione Lens
  • 28. Structure of Retina Fovea centralis Optic Disc Structure of Retina
  • 29. Vitamin A (Retinol) Diet Retinol Intestine Blood stream Liver Retinol binding protein + Retinol Retinal pigmented epithelium Photoreceptors
  • 30. Change in Retinol Retinol Retinene Rhodopsin Retinine reductase (oxidation) Opsin Inside outer segment of photoreceptors
  • 35. Visual Cycle : formation of nerve impulse 1. Rhodopsin changes to metarhodopsin in light 2. Metarhodopsin activates Transducin 3. Transducin activates enzyme phospodiesterase 4. Decreased intracellular cGMP 5. Closure of Na+ channel 6. Hyperpolarisation of rod cells 7. Decreased release of neurotransmitter 8. Response in bipolar cells 9. Signals to optic nerve and then to brain Steps
  • 36. • Collagen is the main content of insoluble proteins in vitreous humor • Hyaluronic acid is glycosaminoglycan. It adds viscosity to the gel. It forms a water binding meshwork around collagen fibers • Soluble proteins has mainly glycoprotein and albumin • Sugar • Ascorbic acid • Amino acids • Electrolyte Vitreous Humor