Zebrafish Science Monitor Vol 4(1)

June 10, 1996

A SOLUTION FOR DELAYED IN VITRO FERTILIZATION OF ZEBRAFISH EGGS

By G. Corley-Smith, Institute of Molecular Biology & Biochemistry, Simon Fraser University, Burnaby, B.C., CANADA

Coho salmon ovarian fluid can be used for delayed in vitro fertilization of zebrafish eggs. Thus, it can be useful for sequential fertilization of small batches of eggs or for procedures where manipulations of eggs prior to fertilization are desired.

Eggs can be squeezed from a female zebrafish, placed in a small amount of coho ovarian fluid, and held at room temperature, with fertilization rates of up to 85% still obtainable after one hour. Various batches of ovarian fluid have varying abilities to preserve zebrafish eggs in a fertilizable state.

To make coho ovarian fluid available to other zebrafish researchers, we have supplied it to a commercial distributor in the USA. We collected coho ovarian fluid in Vancouver, B.C., Canada. We tested individual batches and shipped it to SeaTech Bioproducts, who are now selling it for $20 /ml. This alleviates the difficulties we were experiencing with sending the ovarian fluid through customs to zebrafish labs in the USA.

References below indicate how ovarian fluid is used.

Supplier:
SeaTech Bioproducts
141 California Street
Newton, Massachusetts 02158-1023 USA
Telephone: (617) 965-5092
Voice: (617) 630-5145
e-mail: none yet
www: not yet

Cat # SOF-870-1 ( 1 ml for $20)

Cat # SOF-870-5 (5 ml for $90)

Ovarian fluid is presently shipped by SeaTech on dry ice. As freeze dried samples are much cheaper to mail, we have freeze dried some coho ovarian fluid and will test it shortly for its efficiency in delaying fertilization of zebrafish eggs. We are also working towards developing a defined medium in collaboration with Derek Stemple in Wolfgang Driever's Lab. We have performed an extensive chemical analysis of coho ovarian fluid and hope by the summer of 96 to have a defined medium that has the ability to delay fertilization of zebrafish eggs . At that time we will make the formulation available to the zebrafish community. We presume most researchers would rather buy a relatively cheap mixture than make the mixture from scratch. Thus, if and when we develop such a mixture, we hope a company such as SeaTech will distribute it at a reasonable cost. Until then, coho ovarian fluid is available, at least in a limited supply.

I do not have any affiliation with SeaTech. Collection and testing was performed at the Institute of Molecular Biology and Biochemistry, Simon Fraser University, free of charge so that the coho ovarian fluid could be made available to other zebrafish researchers at as low a fee as possible. Derek Stemple has received and tested coho ovarian fluid. It worked for him.

References:

Corley-Smith, G.E., C.J. Lim and B.P. Brandhorst (1995) Delayed in vitro fertilization using coho salmon ovarian fluid. In The Zebrafish Book - A Guide For The Laboratory Use Of Zebrafish (Danio rerio), edition 3, edited M. Westerfield, Institute of Neuroscience, University of Oregon, pp. 7.22-7.26.

Corley-Smith, G.E., C.J. Lim, and B.P. Brandhorst (1995) Delayed in vitro fertilization of zebrafish eggs using coho salmon (Oncorhynchus kisutch) ovarian fluid. The Zebrafish Science Monitor 3(5): 9-10.

We have a copy of the protocol for use of ovarian fluid on our web site. http://darwin.mbb.sfu.ca/imbb/brandhorst/ofzfbook.htm

Graham Corley-Smith
Institute of Molecular Biology and Biochemistry
Simon Fraser University
Burnaby, B.C.
CANADA
V5A 1S6
Phone: (604) 291-3021
Fax: (604) 291-5583
e-mail: corley@sfu.ca

The Zebrafish Science Monitor, Vol 4(1)

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