NDLI: Transgenic rodent assay for quantifying male germ cell mutant frequency.

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Transgenic rodent assay for quantifying male germ cell mutant frequency.

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Year: 2014 Issue: 89

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Transgenic rodent assay for quantifying male germ cell mutant frequency.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	O'Brien, Jason M. Beal, Marc A. Gingerich, John D. Soper, Lynda Douglas, George R. Yauk, Carole L. Marchetti, Francesco
Description	Author Affiliation: O'Brien JM ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Beal MA ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Gingerich JD ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Soper L ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Douglas GR ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Yauk CL ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre.); Marchetti F ( Environmental Health Science and Research Bureau, Health Canada, Environmental Health Centre)
Abstract	De novo mutations arise mostly in the male germline and may contribute to adverse health outcomes in subsequent generations. Traditional methods for assessing the induction of germ cell mutations require the use of large numbers of animals, making them impractical. As such, germ cell mutagenicity is rarely assessed during chemical testing and risk assessment. Herein, we describe an in vivo male germ cell mutation assay using a transgenic rodent model that is based on a recently approved Organisation for Economic Co-operation and Development (OECD) test guideline. This method uses an in vitro positive selection assay to measure in vivo mutations induced in a transgenic λgt10 vector bearing a reporter gene directly in the germ cells of exposed males. We further describe how the detection of mutations in the transgene recovered from germ cells can be used to characterize the stage-specific sensitivity of the various spermatogenic cell types to mutagen exposure by controlling three experimental parameters: the duration of exposure (administration time), the time between exposure and sample collection (sampling time), and the cell population collected for analysis. Because a large number of germ cells can be assayed from a single male, this method has superior sensitivity compared with traditional methods, requires fewer animals and therefore much less time and resources.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/51576
Journal	Journal of Visualized Experiments
Issue Number	90
Language	English
Publisher	MyJove Corp.
Publisher Date	2014-08-06
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Dna Mutational Analysis Germ-line Mutation Mutation Rate Spermatozoa Physiology Animals Genes, Reporter Mice Mice, Transgenic Cytology Research Support, Non-u.s. Gov't Video-audio Media
Content Type	Text
Resource Type	Article

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