- Publication date : 2016-01-19
Simard O, Leduc F, Acteau G, Arguin M, Grégoire MC, Brazeau MA, Marois I, Richter MV, Boissonneault G. Step-specific Sorting of Mouse Spermatids by Flow Cytometry. J Vis Exp. 2015;:e53379. doi: 10.3791/53379. PubMed PMID: 26780208.
animals cell differentiation dna flow cytometry male mice proteomics spermatids spermatogenesis
The differentiation of mouse spermatids is one critical process for the production of a functional male gamete with an intact genome to be transmitted to the next generation. So far, molecular studies of this morphological transition have been hampered by the lack of a method allowing adequate separation of these important steps of spermatid differentiation for subsequent analyses. Earlier attempts at proper gating of these cells using flow cytometry may have been difficult because of a peculiar increase in DNA fluorescence in spermatids undergoing chromatin remodeling. Based on this observation, we provide details of a simple flow cytometry scheme, allowing reproducible purification of four populations of mouse spermatids fixed with ethanol, each representing a different state in the nuclear remodeling process. Population enrichment is confirmed using step-specific markers and morphological criterions. The purified spermatids can be used for genomic and proteomic analyses.