• Applied Microscopy
• /
• v.27 no.4
• /
• pp.357-372
• /
• 1997

To study the age-related morphological differences of the retinal pigment cells and Bruch's membrane of mouse, retinae of one week-old, five weeks-old, eight weeks-old, six months-old, twelve months-old, eighteen months-old, twenty-four months-old and thirty months-old ICR mice were dissected out under anesthesia. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1 M Millonig's phosphate buffer, pH 7.3), and 1% osmium tetroxide (0.1 M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections were stained with uranyl acetate and lead citrate, and were observed under a JEM 100 CX-II electron microscope. Observed results were as follows: 1. Retinae of one week old mouse exhibit that some parts of the pigment cell provided with basal foldings, whereas other parts of the one contain without basal foldings. After (ive weeks-old, all retinal pigment cells have the basal infoldings. 2. In the one week-old, stage 1 and stage 2 melanosomes were observed in the retinal pigments cells, but after five weeks-old, most of the retinal pigment cells contain some matured stage melanosomes (stage III and stage IV). 3. The phagosomes in the retinal pigment cells were increased during aging. 4. After eighteen months-old, electron dense materials are observed within the basal infoldings. 5. After eighteen months-old, the thickness of the Bruch's membrane is prominently increased. The thickness of the basal laminae of the pigment cell and the choriocapillary endothelium is more prominently increased as compared with that of the other components of the Bruch's membrane. 6. The thickness of the basal lamina of the pigment cell is more prominently increased as compared with that of the choriocapillary endothelium on aging. From the above results, it was suggested that the pigment cell and Bruch's membrane matures structurally In five weeks, and the function of the pigment cell is prominently suppressed around eighteen months-old, and thereafter the functional suppression is continued on aging.

• Applied Microscopy
• /
• v.23 no.2
• /
• pp.11-26
• /
• 1993

This experiment was performed to study the morphological responses of the pigment epithelial cell and the Bruch's membrane of the retina of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80cm, and the. dose rate was 200 rads/min. The experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Under anesthesia, 1% glutaraldehyde-1% paraformaldehyde solution(0.1M Millonig's phosphate buffer, pH 7.3) was perfused through the left ventricle and ascending aorta. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.3) and 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH7.3), and embedded in araldite mixture. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. The morphological changes of the pigment epithelial cells were not pronounced after exposure to 3,000 rads of X-ray. But on the 6th hour after exposure to 6,000 rads of X-ray, bulging nuclear membrane protruding into the cytoplasm and nuclear chromatin clumped into numerous masses along the nuclear membrane were observed. At the 2nd and 6th day post-irradiation, partial cytolysis or necrosis were seen. 2. The thickness of the Bruch's membrane of the experimental groups were increased in the time and dose range covered by this study, and splitting or diffusing basal laminae of the choriocapillary layer were observed frequently in the experimental group. Above results suggest that large amount(6,000 rads) of head irradiation induce direct hazardous effects on the pigment epitherial cells and Bruch's membrane of the retina of the rat, but pigment epithelial cells are more radioresistant than Bruch's membrane.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.6
• /
• pp.1024-1030
• /
• 2007

We investigated the human apolipoprotein E2 (apoE2) transgenic mouse as an animal model system for age-related macular degeneration (AMD). Transgenic mice expressing human apoE2 and C57BL/6J mice were fed normal chow or a high-fat diet for 4 weeks. Eyes were collected from the mice and lipid deposits in retinal pigment epithelium (RPE) were assessed using electron microscopy. The expressions of apoE, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and pigment-epithelium derived factor (PEDF), which are molecular markers for angiogenesis, were assessed with immunohistochemistry. Eyes from apoE2 mice, regardless of diet, contained lipid accumulation in RPE under electron microscopy, whereas control C57BL/6J eyes did not. Lipid accumulation was found predominantly in the RPE and the Bruch's membrane and increased in the eyes of apoE2 mice after one month of a high-fat diet ($8{\pm}2\;per\;50{\mu}m^2$ for normal chow and $11{\pm}2\;per\;50\;{\mu}m^2,\;p<0.05)$. ApoE expression was similar in the apoE2 and control mice; however, VEGF and bFGF were overexpressed in the retinal pigment epithelium of apoE2 eyes compared with control eyes, and PEDF expression was slightly decreased. These expression patterns of VEGF, bFGF, and PEDF suggest angiogenesis is progressing in apoE2 eyes. In conclusion, the eyes of apoE2 mice develop typical lipid accumulations, a common characteristic of AMD, making them a suitable animal model for AMD. The expression profile of VEGF and bFGF on the retinal pigment epithelium suggests that apoE2 may induce neovascularization by altering angiogenic cytokines.