Hostname: page-component-78c5997874-t5tsf Total loading time: 0 Render date: 2024-11-19T22:44:22.939Z Has data issue: false hasContentIssue false
  • English
  • Français

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.
Get access Rights & Permissions [Opens in a new window]

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bernfeld, P. (1951). Enzymes of starch degradation and synthesis. Adv. Enzymol. 12, 379–428.Google ScholarPubMed
Berg, W.E. (1967). Some experimental techniques for eggs and embryos of marine invertebrates. In: Methods in Developmental Biology, ed. F.H., Wilt & N.K., Wessells, pp.767–76. New York: Crowell.Google Scholar
Carroll, E.J., Byrd, E.W. & Epel, D. (1977). A novel procedure for obtaining denuded sea urchin eggs and observations on the role of vitelline layer in sperm reception and egg activation. Exp. Cell Res. 108, 365–74.CrossRefGoogle ScholarPubMed
Chung, H. & Friedberg, F. (1980). Sequence of the N- terminal half of Bacillis amyloliquefaciens alpha amylase. Biochem. J. 185, 387–95.CrossRefGoogle Scholar
Epel, D., Weaver, A.M. & Mazia, D. (1970). Methods for removal of the vitelline membrane of sea urchin eggs. I. Use of dithiothreitol (Cleland reagent). Exp. Cell Res. 61, 64–8.CrossRefGoogle Scholar
Giudice, G. (1986). The Sea Urchin Embryo. Berlin: Springer.CrossRefGoogle Scholar
Kawabe, T.K., Armstrong, P.B. & Pollock, E.G. (1981). An extracellular fibrillar matrix in gastrulating sea urchin embryos. Dev. Biol 85, 509–15.CrossRefGoogle ScholarPubMed
O'Donnell, M.D. & McGeeney, K.F. (1976). Purification and properties of an alpha amylase inhibitor from wheat. Biochim. Biophys. Acta 422, 159–69.CrossRefGoogle ScholarPubMed
Oppenheimer, S.B. & Lefevre, G. (1989). Introduction to Embryonic Development, 3rd edn, pp. 84–113. Boston: Allyn and Bacon.Google Scholar
Schatten, G. & Schatten, H. (1979). Sperm-egg membrane fusions and interactions in denuded sea urchin eggs. Scanning Electron Microsc. 111, 299–305.Google Scholar
Silano, V., Furia, M., Gianfreda, L., Macri, A., Palescandolo, R.A., Scardi, V., Stella, E. & Valfre, F. (1975). Inhibition of amylases from different origins by albumins from the wheat kernel. Biochim. Biophys. Acta 422, 159–69.Google Scholar