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J. Biochem, 1990, Vol. 108, No. 2 193-199
© 1990 Japanese Biochemical Society

research-article

Immunocytochemical Study of the Distribution of a Ganglioside in Sea Urchin Eggs1

Hideo Kubo*,** and Motonori Hoshi*,2

*Department of Life Science, Faculty of Science, Tokyo Institute of Technology Meguro-ku, Tokyo 152
**Developmental and Reproductive Biology Center Yamagata, Yamagata 990

2To whom correspondence should be addressed

An antibody against M5 ganglioside (NeuGca2-6Glcßl-lCer), the dominant ganglioside in the eggs of the sea urchin, Anthocidaris crassispina, was purified by affinity chromatog raphy from rabbit antiserum against crude ganglioside of the eggs. The specificity of the antibody was verified by enzyme-linked immunosorbent assay and TLC iminunostaining. M5 ganglioside was also the major one in the eggs of another sea urchin, Hemicentrotus pulcherrimus, as judged from TLC analyses including immunostalning. Cryostat-sections of H. puicherrimus eggs were examined to determine the distribution of M5 ganglioside by indirect immunofluorescence microscopy with the antibody. Before fertilization, the egg cortex was highly stained, while the other part of cytoplasm was uniformly but much more weakly stained. Mter fertilization, the staining rapidly decreased In the cortex and was restricted to a very thin peripheral layer and to cytoplasmic patches. The Immunore activity was also observed in the esophagus and the somatic cells of the testis, but the spermatozoa were never stained with the antibody.

1This work was supported in part by a Grant-in-Aid for Co-operative Research (No. 61304034), General Scientific Research (No. 61440006), and Scientific Research on Priority Areas (No. 63640003) from the Ministry of Education, Science and Culture of Japan.


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