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Live-cell microscopy of meiosis in spermatocytes.

Journal article
Authors Hiroki Shibuya
Yoshinori Watanabe
Published in Methods in cell biology
Volume 145
Pages 269-277
ISSN 0091-679X
Publication year 2018
Published at Department of Chemistry and Molecular Biology
Pages 269-277
Language en
Links dx.doi.org/10.1016/bs.mcb.2018.03.0...
www.ncbi.nlm.nih.gov/entrez/query.f...
Keywords Animals, Electroporation, methods, Male, Meiosis, physiology, Meiotic Prophase I, physiology, Mice, Microscopy, methods, Spermatocytes, physiology, Testis, physiology, Transgenes, genetics
Subject categories Cell Biology

Abstract

For the analysis of the molecular mechanisms underlying mammalian meiosis, the establishment of a transient gene expression system for meiocytes has been long awaited. We have established an efficient in vivo electroporation method for live mouse testis and demonstrate short-term transgene expression in spermatocytes. By expressing specific marker proteins fused with GFP, this technique is applicable not only to fixed cell observations after transgene expression but also to live imaging to dissect dynamic cellular events in live spermatocytes. The protocol is also adapted to the dissection of mutant phenotypes with defective chromosome movement during meiotic prophase I, as well as a phenotype rescue assay by expressing functional cDNAs in mutant testes.

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