Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes
| Autor: | Jenny Liang, Tierra Strange, Ferdusy Dia, Karen M. Berkowitz, Jacob Hamilton |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Male
0301 basic medicine General Chemical Engineering Meiotic chromosome Biology Chromosomes General Biochemistry Genetics and Molecular Biology Mice 03 medical and health sciences chemistry.chemical_compound Meiosis Spermatocytes Homologous chromosome Animals General Immunology and Microbiology General Neuroscience Chromosome Molecular biology Cell biology Cellular Biology Synaptonemal complex 030104 developmental biology chemistry Homologous recombination Immunostaining DNA |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|