Corneal changes in magnesium-deficient rats

PURPOSE: The purpose of the current study is to investigate the cornea in magnesium (Mg) deficiency and elucidate the local function of trace elements. METHODS: After delivery, mother Wistar Kyoto rats were fed a low Mg diet containing 0.1 mg Mg/100 g diet with all other nutrients and distilled and deionized water. Infant rats were suckled by their mothers for 21 days and then fed the same Mg-deficient diet. Control mother rats were fed commercial rat pellets containing 24 mg Mg/100 g diet and all other nutrients. The corneas were examined by electron microscopy at 6 weeks of age. RESULTS: In the Mg-deficient rats, serum Mg levels were significantly lower and calcium (Ca) levels higher than in the control rats. The corneas of Mg-deficient rats showed decreased microvilli and microplicae in the epithelial cells of the most superficial layer, increased mitochondria with abnormal shapes in the basal cells in the epithelium, condensed chromatin in the nuclei of the basal cells, and high density deposits and macrophage-like cells in the subepithelium of the stroma. Mg-deficient rats had pentagonal and square endothelial cells. CONCLUSION: Since Mg2+ has biologic functions including structural stabilization of protein, nucleic acids, and cell membranes, Mg deficiency may induce changes in the corneal surface and nuclei of corneal epithelial and endothelial cells. These disturbances may interfere with protection from infections, foreign bodies, dryness, and direct exposure to air. Thus, Mg is essential for the cornea to maintain normal structure and function.     

Ultrastructure of the optic nerve in magnesium-deficient rats

PURPOSE: The ultrastructure of the optic nerves in magnesium (Mg)-deficient rats was studied to elucidate the function of Mg. METHODS: After delivery, mother Wistar Kyoto rats were fed a low-Mg diet containing 0.1 mg Mg/100 g diet with all other nutrients and distilled and deionized water. Infant rats were suckled by their mothers for 21 days and then fed the same Mg-deficient diet. Control mother rats were fed commercial rat pellets containing 24 mg Mg/100 g diet and all other nutrients. The optic nerves were examined by electron microscopy at 6 weeks of age. RESULTS: In the Mg-deficient rats, serum Mg levels were significantly lower and calcium levels higher than in the control rats. Ultrastructural findings were: significantly fewer myelinated axons and significantly thinner myelin sheaths in the Mg-deficient rats than in the control and pair-fed controls, and more numerous unmyelinated axons. There were multifocal areas of destruction and necrosis in the optic nerve of Mg-deficient rats. CONCLUSIONS: This study suggests that the optic nerve needs Mg for the development and maintenance of its cell structure.     

The eye and nutrition

Purpose: To examine the effect of vitamins and trace elements on ocular tissue.Materials and Methods: Rats or mice were fed diets deficient in the trace elements Zn, Cu, Mn, Se, Mg, and Cr or in vitamins A, B(12), C, and E. In some rats Al and vitamin A were injected in excessive amounts. We studied the conjunctiva, cornea, retina, and optic nerve with a light microscope, transmission and scanning electron microscopes, an energy dispersive X-ray analyzer, and an ion microscope. Histochemical, cytochemical, and immunohistochemical techniques were applied to the pathological specimens.Results: Deficiencies of Zn, Cu, Mn, and vitamins A, C and E caused a loss of goblet cells in the conjunctiva and a prominent decrease of microvilli and microplicae in the conjunctiva and cornea. The elements in the goblet cells were changed in these conditions. In addition, epithelial cells showed poor fibrous development and abnormal distribution of chromatin in the nucleus.Zn, Cu, Mn, and vitamins A and E deficiencies caused photoreceptor cells to degenerate and disappear. Se deficiency reduced the horizontal and amacrine cells. Vitamin B(12) deficiency reduced nerve fibers in the nerve fiber layer of the retina. Mg deficiency induced multifocal necrosis in the retinal pigment epithelium and apoptotic nuclear changes in the photoreceptor cells. Cr deficiency showed abnormal phagocytosis of the photoreceptor outer segment discs in the retinal pigment epithelium. Vitamin B(12) was found to be related to the circadian rhythm in the retina.Deficiencies of Zn, Cu, Mn, and vitamins A, B(12), and E induced degeneration and disappearance of myelin lamellae in the myelinated optic nerve fibers.In hypervitaminosis A, lipid droplets appeared in the retinal pigment epithelium and alcohol dehydrogenase disappeared in the retinal pigment epithelium and photoreceptor outer segments. Excessive Al was toxic to the retina, which showed disappearance of photoreceptor cells. Al deposits were seen in dendrites and neurons in the outer plexiform layer.Zn seemed to be necessary for corneal epithelial cell wound healing.Discussion: Trace elements usually are contained in enzymes, which have many metabolic functions. They are related to synthesis and breakdown of many substances. Some trace elements such as Zn, Cu, Mn, and Se and vitamins including vitamins A, C, and E prevent peroxidation of lipids. Some vitamins have an affinity for specific tissues such as epithelial cells, nerve fibers, and neuronal cells and are needed for cell differentiation, development, and maintenance.Conclusion: Cu, Zn, Mn, Se, Mg, and Cr and vitamins A, B(12), C, and E are necessary for maintenance of cellular structure and metabolism.     

The eye and nutrition

PURPOSE: To examine the effect of vitamins and trace elements on ocular tissue. MATERIALS AND METHODS: Rats or mice were fed diets deficient in the trace elements Zn, Cu, Mn, Se, Mg, and Cr or in vitamins A, B12, C, and E. In some rats Al and vitamin A were injected in excessive amounts. We studied the conjunctiva, cornea, retina, and optic nerve with a light microscope, transmission and scanning electron microscopes, an energy dispersive X-ray analyser, and an ion microscope. Histochemical, cytochemical, and immunohistochemical techniques were applied to the pathological specimens. RESULTS: Deficiencies of Zn, Cu, Mn, and vitamins A, C and E caused a loss of goblet cells in the conjunctiva and a prominent decrease of microvilli and microplicae in the conjunctiva and cornea. The elements in the goblet cells were changed in these conditions. In addition, epithelial cells showed poor fibrous development and abnormal distribution of chromatin in the nucleus. Zn, Cu, Mn, and vitamins A and E deficiencies caused photoreceptor cells to degenerate and disappear. Se deficiency reduced the horizontal and amacrine cells. Vitamin B12 deficiency reduced nerve fibers in the nerve fiber layer of the retina. Mg deficiency induced multifocal necrosis in the retinal pigment epithelium and apoptotic nuclear changes in the photoreceptor cells. Cr deficiency showed abnormal phagocytosis of the photoreceptor outer segment discs in the retinal pigment epithelium. Vitamin B12 was found to be related to the circadian rhythm in the retina. Deficiencies of Zn, Cu, Mn, and vitamins A, B12, and E induced degeneration and disappearance of myelin lamellae in the myelinated optic nerve fibers. In hypervitaminosis A, lipid droplets appeared in the retinal pigment epithelium and alcohol dehydrogenase disappeared in the retinal pigment epithelium and photoreceptor outer segments. Excessive Al was toxic to the retina, which showed disappearance of photoreceptor cells. Al deposits were seen in dendrites and neurons in the outer plexiform layer. Zn seemed to be necessary for corneal epithelial cell wound healing. DISCUSSION: Trace elements usually are contained in enzymes, which have many metabolic functions. They are related to synthesis and breakdown of many substances. Some trace elements such as Zn, Cu, Mn, and Se and vitamins including vitamins A, C, and E prevent peroxidation of lipids. Some vitamins have an affinity for specific tissues such as epithelial cells, nerve fibers, and neuronal cells and are needed for cell differentiation, development, and maintenance. CONCLUSION: Cu, Zn, Mn, Se, Mg, and Cr and vitamins A, B12, C, and E are necessary for maintenance of cellular structure and metabolism.     

Light treatment enhances photoreceptor survival in dystrophic retinas of Royal College of Surgeons rats.

PURPOSE: To determine whether treatment with bright light elicits a protective response that enhances photoreceptor survival in Royal College of Surgeons (RCS) rats with inherited retinal degeneration. METHODS: RCS rats were illuminated for 10 to 12 hours with 130 foot-candles (fc) of white or green light. Untreated littermates that were kept under low cyclic light levels were used as control subjects. Photoreceptor survival was determined by quantitative analysis of photoreceptor nuclei and ultrastructural assessment of cellular organization. Basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF) gene expression were determined at the mRNA and protein levels. RESULTS: Treatments of RCS rats with a single dose of bright light on postnatal day 23 (P23) greatly enhanced photoreceptor survival. Ultrasturctural analysis revealed intact inner segments in light-treated retinas, whereas in untreated retinas only remnants of inner segments were observed. By P42, numerous viable nuclei were counted in the posterior retina of light-treated rats, whereas most of the remaining nuclei in untreated RCS rat retinas were highly pyknotic. At 2.5 days after treatment with a single dose of bright light, bFGF gene expression was significantly higher than in untreated RCS rat retinas. By P42, bFGF protein levels were still significantly higher in the treated retinas. CONCLUSIONS: Exogenous bFGF has been shown to promote photoreceptor survival in the RCS rat retina. Thus, the increased bFGF expression that was measured in the light-treated RCS rat retinas may be a protective response to light stress, which supports the observed rescue of photoreceptors in light-treated RCS rat retinas.     

Retinal light damage: Protective effect of α-tocopherol

We studied histologically the protective effect of α-tocopherol to retinal light damage. After 3-week-old albino rats were fed with an α-tocopherol deficient or supplemented diet and kept in a 12-hour dim light/12-hour dark environment for 8 weeks, each animal was exposed to intense light (2500 lux) for 1, 3, 6, 12, 24, and 72 hours. The eyes were enucleated and prepared for transmission electron microscopy study and image analysis of phagosomes. Before light exposure, the α-tocopherol content of the neural retina of the deficient and supplemented groups was 0.3 μg and 23.34 μg, respectively. After 1- and 3-hour exposures, morphological changes in the retinal pigment epithelium and photoreceptor membranes were more extensive in the deficient group than in the supplemented group. After a 24-hour exposure, pyknotic photoreceptor nuclei were more numerous in the deficient group than the supplemented group. After 3-, 6-, and 12-hour exposures, large phagosomes were more numerous in the deficient group than in the supplemented group. These findings suggest that α-tocopherol can protect the retina from light injury for up to 24 hours of exposure.     

Age-related and light-associated retinal changes in Fischer rats

Morphological changes in retinas of aging Fischer 344 rats were characterized. The numbers of photoreceptor cells gradually decreased as rats aged. The outer nuclear layer was 12 cells thick at 3 months, but was reduced to less than 8 cells by 18 months. The decrease of photoreceptor cells was more pronounced in rats housed under a light intensity of 32-ft-c than in rats housed under a light intensity of 1 ft-c. Inner and outer segments of surviving photoreceptor cells were morphologically normal. A new form of retinal degeneration was discovered in aged Fischer rats characterized by selective degeneration of peripheral retina. Degeneration was characterized by severe loss of photoreceptor cells in the far peripheral retina. Microcystoids were found in about 25% ofthe affected retinas, and the loss of photoreceptor cells was followed by proliferation and vascularization of the retinal pigment epithelium and disorganization of retinal structures. The incidence and severity of peripheral retinal degeneration increased with aged and prolonged exposure to comparatively high-intensity light. All Fischer rats ((5/5) housed under light intensity of 32 ft-c developed severe peripheral retinal degeneration by 24 months. Peripheral retinal degeneration was an age-related change but appeared to be exaggerated by ambient light.     

Vitamin E deficiency and the retina: photoreceptor and pigment epithelial changes.

To investigate the role of normal vitamin E levels and the interrelationships between vitamin E and A in maintaining the visual cells of the retina, weanling female Sprague-Dawley rats were fed vitamin E-free diets differing tenfold in their vitamin A content (0.8 and 8.0 mg of retinol per kilogram of diet). Rats on vitamin E-free diets with the higher vitamin A level exhibited marked disruption of photoreceptor outer segment membranes and a fivefold increase in the number of lipofuscin granules in the pigment epithelial cells which ingest these membranes. Rats on vitamin E-free diets with the lower vitamin A level showed the same retinal damages plus significant loss of photoreceptor cells compared to age-matched rats on control diets. Rods and cones were involved equally, and their pattern of loss was not like that found in vitamin A deficiency. Normal levels of vitamin E probably protect photoreceptor membranes from oxidative damage and retard the accumulation of their remnants and other products of lipid breakdown in the pigment epithelium. The vitamin A status of rats has a significant influence on the extent of damage induced by vitamin E deficiency.     

Retinal morphology in rats treated with a taurine transport antagonist

Previous studies have shown that chronic treatment of rats with guanidinoethyl sulfonate (GES), an antagonist of taurine uptake, leads to depletion of retinal taurine content and electroretinogram abnormalities. In this study we examined with the light- and electron microscope the retina and pigment epithelium of mother rats treated for 8 weeks with 1% GES in their drinking water. Pigment epithelial cells appeared swollen, sometimes with mitochondrial vacuolization and loss of cytoplasmic density, and changes in nuclear shape and chromatin distribution. Photoreceptor cells had shorter outer segments with disarray of disks, swollen inner segments and nuclei, and loss of synaptic terminal contents. These degenerative changes frequently led to cell death, indicated by loss of photoreceptor cell nuclei and shrinkage of the outer nuclear layer, and disappearance of photoreceptor synaptic terminals from the outer plexiform layer. Narrowing of the outer plexiform layer was due also to the retraction of cell processes from higher-order neurons. Quantitative studies of the retina along the vertical meridian passing through the optic nerve head showed that treatment resulted in significant reductions in the number of rod-cell nuclei per field, the width of the outer nuclear layer, and the width of the inner retina. Effects were more pronounced in the superior than the inferior hemisphere, reminiscent of findings in light-damage studies. A possible role for taurine in the cellular defence mechanisms against light- and oxygen-induced damage is suggested.     

Maintenance of opsin density in photoreceptor outer segments of retinoid-deprived rats

Dietary deficiency in the retinoid precursors of the visual pigment chromophore 11-cis-retinal eventually results in selective degeneration of the photoreceptor cells of the vertebrate retina. Early effects of retinoid deficiency are depletion of rhodopsin from the retina and vesiculation of the photoreceptor outer segment disc membranes. Experiments were conducted to determine whether these early changes were accompanied by an alteration of the opsin content of the disc membranes. After being fed a retinoid-deficient diet containing retinoic acid for 26 weeks, the rhodopsin content of rat retinas was reduced by over 85%. Both the diameters and the lengths of the outer segments decreased significantly. However, immunocytochemical and freeze-fracture analyses indicated that retinoid deficiency did not lower opsin density in the outer-segment disc membranes. These findings indicate that in the rat, opsin synthesis and disc assembly are coordinated processes that remain coupled despite reduced availability of the vitamin A chromophore. The fact that disc size decreases and disc synthesis eventually ceases in retinoid-deprived rats indicates that specific retinoids are essential for disc morphogenesis. The mechanism by which these retinoids regulate disc assembly remains to be determined.     

A morphological study of the acute toxicity of l-cysteine on the retina of young rats

Systemic administration of l-cysteine to rats (1·2 mg/g body weight) during their first ten days of life leads to a permanent retinal dystrophy which affects the inner layers of retina. To clarify which types of retinal cells that are involved in this lesion, 9- to 10-day-old rats were injected subcutaneously with l-cysteine. The retinas obtained from these animals 4 hr later were examined by light and electron microscopy. Numerous amacrine cells with pyknotic nuclei and swollen cytoplasm were observed. Between these cells undergoing necrosis, well preserved amacrine cells were found. Similar changes were found in about 50% of the ganglion cells. No specific changes were observed in the bipolar cells, the Müller cells or the horizontal cells. The photoreceptors and the pigment epithelium were unaffected. It is concluded that a sublethal dose of l-cysteine given systemically to young rats (up to the age of 10 days), affects subpopulations of the amacrine and the ganglion cells of the retina. Potentially, l-cysteine used in this way, may be a useful scientific tool to destroy selectively populations of retinal neurons.     

Structural and biochemical changes in vitamin A--deficient rat retinas.

The levels of rhodopsin and opsin were investigated in relation to the maintenance of retinal structure in retinas of vitamin A--deficient rats in low levels of cyclic illumination (1.5 to 2 foot-candles). Rhodopsin levels decreased in the deficient retinas to approximately 20% of control at 9 weeks, and this level was retained through 39 weeks on the deficient diet. Opsin levels decreased at a slower rate but reached about 20% of control levels at 32 weeks. Despite the decrease in rhodopsin levels, obvious deterioration of disc structure was not observed until 16 weeks of deficiency, when opsin levels had already decreased to 60% to 70% of control. The structural disruption of photoreceptor outer segments was localized initially in discs of the distal third. Rod cell degeneration preceded cone cell degeneration in vitamin A--deficient retinas. Most of the rods and cones persisted in the posterior retina at 23 weeks on the deficient diet; however, by 40 weeks, only 11% of the rod nuclei remained. In contrast, about 63% of the cone nuclei were present at 40 weeks of deficiency. The photoreceptor cells were affected by the deficiency to a greater extent in the inferior hemisphere than in the superior hemisphere of the eye.     

The effect of methylazoxymethanol acetate (MAM) on the developing rat retina

The morphological effects of methylazoxymethanol acetate (MAM) on the prenatal and postnatal rat retina were studied using light and electron microscopy. MAM was injected into pregnant rats once on days 9(E9) to 20(E20) of gestation, or into newborn pups once on days 1(PE1) through 9(PE9) after birth. Some of the retinas were examined 3–4 weeks after birth and others at 5 months. A second group of rats was injected with MAM on day PE1 and killed at 24-h intervals to day PE9. The retina from adult animals treated with MAM between days E17 and PE5 (except day E20) showed pathological damage. The retinas from adult animals treated with MAM before E17 and after PE5 appeared normal. Retinas from animals examined daily after injection of MAM had, within 2 days of the treatment, pyknotic cells and necrosis in the neuroblastic layer. By 5 days after treatment no necrotic material was visible. At 3 days after treatment, dark cells, believed to be aberrant developing photoreceptor cells, were adjacent to the pigment epithelium. These cells appeared to be the focal point for formation of rosettes which occurred on subsequent days.The developing rat retina is sensitive to MAM during gestation and the early postnatal period. Before E17 when most of the cells were still in the proliferating pool, the retina may have been able to repair the MAM-induced injury and develop normal cytoarchitecture. Between E17 and PE6 when more and more cells became postmitotic the retina was not able to compensate and rosettes were formed. Finally, after PE6 when most cells were postmitotic and less were killed by MAM, the retina again appeared not to be affected.     

Retinal dystrophy in Wistar-Furth rats

A form of retinal degeneration in the Wistar-Furth strain of rat is described. The changes observed was classified as either mild or severe, depending on the severity of the dystrophy seen by light and electron microscopy. In the mild stage, there was a slight decrease in the number of photoreceptor nuclei in the outer nuclear layer (ONL) and there were pyknotic nuclei in the inner nuclear layer. A few photoreceptor outer and inner segments showed focal swellings. The basal infoldings of the retinal pigment epithelium (RPE) were distributed irregularly, creating spaces that contained collagen-like fibrils. However, the interepithelial tight junctions remained impermeable to intravenously injected peroxidase. In severely dystrophic retinas, there was a further decrease in the width of the ONL, and the photoreceptor outer and inner segments were degenerated. In extreme cases, the photoreceptor cell disappeared completely and the inner retina abutted the RPE. Further changes were also noted in the basal membrane of the RPE; in some regions the membrane was totally flat while in others it formed exaggerated infoldings. In addition, the RPE was frequently vascularized by vessels that originated from the retina. The endothelium of those intra-RPE vessels located near Bruch's membrane was frequently fenestrated. Alterations also occurred in the retinal capillaries; some were degenerated, while others showed increased tortuosity or focal thickenings of the basal lamina. In addition, there were focal increases in the amount of stroma between Bruch's membrane and the choriocapillaris.     

Caspase activation in an experimental model of retinal detachment

PURPOSE: To test for apoptotic photoreceptor cell death and caspase activation as a function of time after induction of an experimental retinal detachment. METHODS: Retinal detachments were created in Brown Norway rats by injecting 10% hyaluronic acid into the subretinal space using a transvitreous approach. Light microscopy and terminal dUTP-biotin nick end-labeling (TUNEL) was performed at 1, 3, 5, and 7 days after detachment to assess for the morphologic features associated with apoptosis. Western blot analysis of retinal protein extracts was performed using antibodies against caspase-3, -7, and -9 and poly-ADP ribose-polymerase (PARP) at 1, 3, and 5 days after detachment. RESULTS: Light microscopic analysis of detached retinas showed the presence of pyknotic nuclei in the outer nuclear layer and disruption of the normal organization of the photoreceptor outer segments. TUNEL-staining was positive in the outer nuclear layer only in the detached portions of the retina. Western blot analysis confirmed the time-dependent activation of caspase-3, -7, and -9 and PARP in the detached retinas. No morphologic stigmata of apoptosis or caspase activation was detected in attached retinas. CONCLUSIONS: The apoptotic photoreceptor cell death in experimental retinal detachments is associated with caspase activation.     

Retinal changes in Otsuka long-evans Tokushima Fatty rats (spontaneously diabetic rat)--possibility of a new experimental model for diabetic retinopathy

PURPOSE: To study retinal changes in the Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a spontaneously diabetic rat, and evaluate it as a possible new diabetic retinopathy model. METHODS: We examined the retinas of OLETF rats and Long-Evans Tokushima Otsuka (LETO) rats as controls under both a transmission electron microscope (TEM) and a scanning electron microscope (SEM). RESULTS: We detected higher blood sugar level in the OLETF rats than in the LETO rats beginning at 5 months of age. The inner nuclear layers of the retina decreased from 3-4 rows to 2 rows, whereas the photoreceptor cell nuclei decreased from 8 rows to 3-6 rows. Retinal pigment epithelial cells decreased in height, and basal infoldings were poorly developed. Retinal capillary basement membranes were significantly thicker in the OLETF rats than in the LETO rats, and endothelial cell damage was observed. The SEM of vascular corrosion casts of OLETF rats showed tortuosity, microaneurysms, and loop formations. CONCLUSIONS: The development of spontaneous hyperglycemia in OLETF rats was associated with alterations in retina ultrastructure. Changes were similar to those seen in diabetic retinopathy, but did not include either hemorrhages or exudates. The OLETF rat may be a useful animal model for the study of ocular complications in humans.     

Transplantation of photoreceptors to light-damaged retina

We investigated the possibility of reconstructing light-damaged retinas by photoreceptor transplantation. The outer nuclear layer containing the photoreceptor cells was eliminated in adult albino rat retinas by exposure to constant high-level illumination. Photoreceptors for transplantation were harvested from neonatal rats using a novel isolation technique that maintained the cellular organization of the outer nuclear layer. Transplantation was accomplished using a transcorneal approach to the subretinal space, which minimized trauma to the eye. The retina reattached to the back of the eye with transplanted photoreceptors interposed between the retina and the overlying tissues. Prelabelling with fluorescent dye enabled positive identification of the transplanted cells. The transplanted photoreceptors appeared to survive transplantation for at least 6 weeks and were immunohistochemically reactive for opsin. The antibody staining for opsin identifies the transplanted cells as photoreceptors and indicates that they are still capable of producing visual pigment and therefore may have the capacity to transduce light. These findings indicate that photoreceptors can be transplanted to form a new outer nuclear layer in a damaged mature retina.     

Immunohistochemical localization of basic fibroblast growth factor in mature and developing retinas of normal and RCS rats.

Basic fibroblast growth factor (bFGF) delays photoreceptor degeneration when injected intraocularly in Royal College of Surgeons (RCS) rats with inherited retinal dystrophy. In the present study, we have determined the localization of endogenous bFGF in retinas of normal and RCS rats during the normal developmental period (postnatal days 0-20) and the period of photoreceptor degeneration in RCS rats (days 20-90). bFGF was localized immunohistochemically by indirect immunoperoxidase using two different polyclonal antibodies and one monoclonal antibody against bFGF. bFGF was present in retinas as early as birth, and remained through adult age. Controls using either PBS, non-immune IgG or antibody preabsorbed with bFGF peptide were devoid of label. In normal rats between the ages of birth and postnatal day (P) 4, bFGF was found in developing ganglion cells, superficial blood vessels, some of the innermost cells in the neuroblastic layer, developing horizontal cells, and retinal pigment epithelial (RPE) cells. Between P0 and P4, the intensity of staining increased significantly in horizontal cells. From P6-P10, some cells in the inner nuclear layer remained positive, but horizontal cell staining became less intense in the central retina. The superficial vessels, ganglion cells and RPE cells also remained positive for bFGF. At P20-25, when the retina was essentially mature, bFGF was found in RPE cells, most cells of the ganglion cell layer, and many cells of the inner nuclear layer, but horizontal cells and blood vessels showed a lower concentration of bFGF than they did at younger ages. At P45 and older, blood vessels no longer showed bFGF immunoreactivity. The staining pattern in RCS rats was indistinguishable from that for normal rats at all ages examined. These results show that bFGF is present in the developing and adult rat retina in some neural cells, in addition to vessels and RPE cells. The transient elevated expression of bFGF immunoreactivity in developing horizontal cells and blood vessels suggests a possible role for this growth factor in retinal development. In addition, if RCS retinas possess any difference in bFGF localization or concentration compared to normal retinas, it must be too small to detect by immunohistochemical means, or at least with the reagents used.