ANALYSIS OF THE MECHANISM OF CYTOKINESIS SPECIFIC FOR EMBRYONIC CELLS.
Project/Area Number |
04670028
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Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
General anatomy (including Histology/Embryology)
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Research Institution | KANSAI MEDICAL UNIVERSITY |
Principal Investigator |
ASADA Mari KANSAI MEDICAL UNIVERSITY, FACULTY OF MEDICINE, LECTURER, 医学部, 講師 (60121901)
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Project Period (FY) |
1992 – 1993
|
Project Status |
Completed (Fiscal Year 1993)
|
Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥1,400,000 (Direct Cost: ¥1,400,000)
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Keywords | Cell division / Cleavage furrow / Two-dimensional gel-electrophresis / Maternal-effect mutation / Monoclonal antibody / Western blotting analyzes / Xenopus laevis / Wheat germ agglutinin / 卵割 |
Research Abstract |
We have isolated maternal-effect mutant (af) females of Xenopus laevis by the back-cross of a female with one of her sons. The af females lay eggs whose cytoplasmic divisions do not take place while nuclear divisions are not affected.The purpose of the present study is to investigate the molecular mechanism of cytokinesis specific for embryonic cells by the use of af mutant eggs. The following results were obtained. 1.Two-dimensional gel electrophoretic analysis showed that a 78 kDa (pI 7.6) spot which was present in wild-type egg cytoplasm was completely missing in af egg cytoplasm. The N-terminal 20 amino acid sequence of the 78-kDa protein had no homology to those of known proteins. 2.A monoclonal antibody against the 78-kDa protein was raised. Western blotting analyzes showed that the anti-78 kDa antibody specifically rocognized a set of two or three bands in eggs and embryos of wild-type Xenopus, but only one band in those of af. 3.As the molecular masses of a set of bands were similar to those of ezrin-radixin-moesin (ERM) family proteins which are konwn to link actin-based cytocketeton to the plasma membrane, we also analyzed the ERM family proteins by Western blotting. The results showed that the 78 kDa protein was not identical to any of the ERM family proteins. 4.As the wild-type eggs treated with wheat germ agglutinin (WGA) showed cleavage arrest and the morphlogy of the arrested eggs was very similar to that of af eggs, we compared WGA-binding proteins between wild-type and af eggs. The results showed that a 205-kDa protein was absent in af embryos and a 34-kDa protein was also affected by af mutation. We are currently investigating the roles of these proteins in cytoplasmic divisions.
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Report
(3 results)
Research Products
(5 results)