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Producing exposed cota-free embryos

Published online by Cambridge University Press:  26 September 2008

Michael F. Daily ,
Virginia H. Latham ,
Claudia M. Garcia ,
Cynthia L. Hockman ,
Helen Chun ,
Mark L. Oppenheimer ,
Steven P. West ,
Karolin Rostamiany ,
Richard L.C. Chao  and
Edward G. Pollock
...Show all authors
Michael F. Daily
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Virginia H. Latham
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Claudia M. Garcia
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Cynthia L. Hockman
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Helen Chun
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Mark L. Oppenheimer
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Steven P. West
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Karolin Rostamiany
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Richard L.C. Chao
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Edward G. Pollock
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
Steven B. Oppenheimer*
Affiliation:
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge, California, USA.
*
Steven B.Oppenheimer, Department of Biology and Center for Cancer and Developmenatal Biology, California State University, Northridge, 18111 Nordhoff Street, Northridge, CA 91330-8303, USA. Tel: 818/885-3336. Fax: 818/717-4030.
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Summary

Production of embryos that are free of tough outer coats facilitates studies that are not possible with embryos surrounded by impenetrable envelopes. This report describes a new procedure for preventing formation of fertilisation membranes in the sea urchin (Lytechinus pictus) model. This procedure involves treating unfertilised eggs with the enzyme alpha-amylase, which cleaves alpha-1,4 glucosidic bonds in the vitelline layer. A major advantage of this method is that it is very well defined and completely controllable with alpha-amylase inhibitor. The results suggest that intact alpha-1,4 glucosidic bonds are essential for vitelline layer integrity required for formation of the fertilisation membrane. Eggs treated with alpha-amylase possessed the same surface lectin receptors as untreated eggs and, as shown by light and transmission electron microscopy, produced healthy, cleaving embryos that were free of fertilisation envelopes.

Keywords

Fertilisation membraneSea urchinVitelline
Type
Article
Information
Zygote , Volume 2 , Issue 3 , August 1994 , pp. 221 - 225
Copyright
Copyright © Cambridge University Press 1994

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