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Journal article
Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, v 2017(129)
01 Nov 2017
PMID: 29286440
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Mammalian meiosis is a dynamic developmental process that occurs in germ cells and can be studied and characterized. Using a method to spread nuclei on the surface of slides (rather than dropping them from a height), we demonstrate an optimized technique on mouse spermatocytes that was first described in 1997. This method is widely used in laboratories to study mammalian meiosis because it yields a plethora of high quality nuclei undergoing substages of prophase I. Seminiferous tubules are first placed in a hypotonic solution to swell spermatocytes. Then spermatocytes are released into a sucrose solution to create a cell suspension, and nuclei are spread onto fixative-soaked glass slides. Following immunostaining, a diversity of proteins germane to meiotic processes can be examined. For example, proteins of the synaptonemal complex, a tripartite structure that connects the chromosome axes/cores of homologs together can be easily visualized. Meiotic recombination proteins, which are involved in repair of DNA double-strand breaks by homologous recombination, can also be immunostained to evaluate progression of prophase I. Here we describe and demonstrate in detail a technique widely used to study mammalian meiosis in spermatocytes from juvenile or adult male mice.
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Details
- Title
- Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes
- Creators
- Ferdusy Dia - Drexel UniversityTierra Strange - Drexel UniversityJenny Liang - Drexel UniversityJacob Hamilton - Drexel UniversityKaren M. Berkowitz - Drexel University
- Publication Details
- JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, v 2017(129)
- Publisher
- Journal Of Visualized Experiments
- Number of pages
- 5
- Grant note
- R01GM106262 / NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of General Medical Sciences (NIGMS) R01 GM106262 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000417688700005
- Scopus ID
- 2-s2.0-85037627280
- Other Identifier
- 991019169675804721
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- Web of Science research areas
- Multidisciplinary Sciences