(none)

Graham Corley-Smith corley at sfu.ca
Sun Feb 18 19:32:30 EST 1996


Hi All,
LONG MESSAGE:  CONCERNS AVAILABILITY OF A SOLUTION USEFUL FOR DELAYED IN
VITRO FERTILIZATION OF ZF EGGS.

     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 zf 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 media
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 media 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.  Till 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 zf 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, pp. 7.22 - 7.26 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.

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 at sfu.ca 


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 at sfu.ca 





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