title: Step-specific Sorting of Mouse Spermatids by Flow Cytometry.
abstract: 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.
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Olivier
authors.initials: O
authors.lastname: Simard
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Leduc
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Acteau
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Mélina
authors.initials: M
authors.lastname: Arguin
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Marie-Chantal
authors.initials: MC
authors.lastname: Grégoire
authors.affiliation: Department of Biochemistry, Université de Sherbrooke.
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Brazeau
authors.affiliation: Department of Medicine, Université de Sherbrooke.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Marois
authors.affiliation: Department of Medicine, Université de Sherbrooke.
authors.forename: Martin V
authors.initials: MV
authors.lastname: Richter
authors.affiliation: Department of Biochemistry, Université de Sherbrooke; Guylain.Boissonneault@usherbrooke.ca.
authors.forename: Guylain
authors.initials: G
authors.lastname: Boissonneault
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D008297:Male; D051379:Mice; D040901:Proteomics / Q000379:methods; D013087:Spermatids / Q000737:chemistry / Q000145:classification* / Q000166:cytology; D013091:Spermatogenesis
chemical_list: D004247:DNA