Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
|Research Institution||Yamagata University|
TONASAKI Akira Yamagata University school of medicine, Professor, 医学部, 教授 (90004572)
MORI Keiji Yamagata University school of medicine, Assistant, 医学部, 助手 (40250914)
SASASKI Katsunori Yamagata University school of medicine, Associate Professor, 医学部, 助教授 (30170666)
TOKENAGA Fumio Osaka University, Faculty of Science, Professor, 理学部, 教授 (80025452)
|Project Period (FY)
1994 – 1996
Completed(Fiscal Year 1996)
|Budget Amount *help
¥29,400,000 (Direct Cost : ¥29,400,000)
Fiscal Year 1996 : ¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1995 : ¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1994 : ¥25,000,000 (Direct Cost : ¥25,000,000)
|Keywords||retina / lamprey / visual pigment / monoclonal antibody / rhodopsin / arrestin / immmunohistochemisity / 電顕免疫組織化学 / 光受容細胞の分化 / 光受容細胞 / モノクローン抗体 / 免疫電顕組織化学|
1.Production of monoclonal antibodies against lamprey-retinal homogenate.
We have developed 10 new monoclonal antibodies. Their binding sites were detected on the sectioned retina by optical and electron microscopic immunohistochemistry. Molecular weight of antigens were estimate by western blotting.
1) W26, W27, W39 : Specifically bound to the outer segment of short photoreceptor cell (SPC).
2) W226, W231, W312, W425 : To the outer segment, inner segment except for ellipsoid of SPC and axon.
3) W15 : To the outer and inner segment of SPC.
4) W38 : To the all region of SPC.
5) W318 : To the neural layr of retina.
Following 3 monoclonal antibodies were specialized for a molecular component involved in photoreceiving procedure.
1) W39 : For lamprey rhodopsin, molecular weight is about 40kDa.
2) W312, W425 : For lamprey arrestin, molecular weight is about 49kDa.
2.Establishment of quantitative immunogold electron microscopy.
The specimen was fixed with 4% paraformaldehyde and embedded in LR-white. There were considerably remakable washing out and trace of destruction in the neural layr of retina, and deformation of outer segment of photoreceptor cells as compared with conventional fixation. As far as photoreceptor cells are concerned, however, fortuitously preserved outer segments were found as assosiated variously with fine gold particles, allowing our quantitative examinatoin.
A series of anti-lamprey retinal antibodries were produced and used for recognition of protein, glycoconjugate and lipids in retinal cells. These antibodies are apparently useful as probes for relationship between corresponding cells of different species.