Making gynogenetic diploid zebrafish by early pressure.

Walker, Charline and Walsh, Greg S and Moens, Cecilia (2009) Making gynogenetic diploid zebrafish by early pressure. Journal of visualized experiments : JoVE (28). p. 1396. ISSN 1940-087X

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Abstract

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al. described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book.

Item Type:	Article or Abstract
DOI:	10.3791/1396
PubMed ID:	19568220
NIHMSID:	NIHMS137495
PMCID:	PMC2798835
Keywords or MeSH Headings:	* Animals * Diploidy* * Female * Genetic Techniques * Homozygote * Male * Ovum/physiology * Spermatozoa/physiology * Spermatozoa/radiation effects * Ultraviolet Rays * Zebrafish/genetics*
Subjects:	Cellular and Organismal Processes > Genetic processes Organisms > Model organisms Molecules > Chromosomes
Depositing User:	Library Staff
Date Deposited:	03 Aug 2009 23:35
Last Modified:	14 Feb 2012 14:42
URI:	http://authors.fhcrc.org/id/eprint/324

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