|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1993 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1992 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1991 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Using the technique of two-dimensional electrophoresis, more than 30 clusters of over 80 spots were detected in the vitreous. In addition to 13 serum proteins, a number of proteins not found in serum were detected by comparison with the electro phoretic pattern of serum. The most significant one had 47,000 molecular weight(MW), migrating slightly to the acidic side of albumin. The others included spots with 88,000, 25,000, 21,000 and 17,000 MW.
To confirm that these are non-serum proteins, affinity chromatography using anti-human whole serum was performed. Bound and unbound fractions of vitreous samples represented approximately 80% and 20% of total protein recovered. The electrophoretic pattern of the bound fraction showed the presence of several major serum proteins. The most of proteins not detected in the electrophoretic pattern of serum were detected in that of the unbound fraction.
In order to know the postmortem changes, electrophoretic patterns of rabbit vitreous samples obtained
immediately and 48 hours after death were compared, and at least in 48 hours after death, any spots not found in the immediate sample was not detected.
The unfractionated soluble protein of the vitreous demonstrated significant stimulation of retinal pigment epithelial(RPE) cell growth in culture showing 207.8% compared with control(100%). The bound(serum) and the unbound(non-serum) fractions also stimulated proliferation of RPE cells, showing 156.5% and 159.7% respectively. Although no significant differences between stimulatory activities of these fractions were present, the non-serum fraction was approxi mately 4 times more stimulatory than the serum fraction when corrections were made for protein content.
These results suggest that there are more proteins in the vitreous than have been previously reported and that some of these may be of non-serum origin. Also suggested was that the soluble proteins of the vitreous stimulate proliferation of RPE cells and may participate in the pathogenesis of proliferative vitreoretinopathy. Less