title: Analysis of the complexity of the sperm acrosomal membrane by super-resolution stimulated emission depletion microscopy compared with transmission electron microscopy.
abstract: The acrosome is a Golgi-derived sperm cell organelle enclosed by a continuous acrosomal membrane. The acrosomal membrane complexes with surrounding matrices containing molecules necessary for fertilization; however, the complex of acrosomal membrane and associating matrices (CAMAM) has not been visualized in detail under living conditions. Here, we analyzed the CAMAM at the nanometer level using super-resolution stimulated emission depletion (STED) fluorescence microscopy and equatorin-enhanced green fluorescent protein transgenic mice. The STED images were compared with the corresponding images taken by immunoelectron microscopy. Consequently, the substructure of CAMAM could be differentiated at nanometer-scale resolution by STED microscopy without the need for sectioning. The information obtained in this study will be beneficial not only for understanding the molecular mechanism of fertilization but also for cell imaging under living conditions.
authors.affiliation: Department of Reproductive Biology and Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan chizuru@faculty.chiba-u.jp.
authors.forename: Chizuru
authors.initials: C
authors.lastname: Ito
authors.affiliation: Department of Reproductive Biology and Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan Biomedical Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamatoya
authors.affiliation: Department of Reproductive Biology and Medicine, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Toshimori
mesh_terms: D000177:Acrosome / Q000502:physiology*; D020101:Acrosome Reaction / Q000502:physiology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002462:Cell Membrane; D005306:Fertilization; D049452:Green Fluorescent Proteins / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013194:Staining and Labeling; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; C512717:Eqtn protein, mouse; D008565:Membrane Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: CLSM; Eqtn-EGFP transgenic mouse; STED microscopy; complexity of sperm acrosomal membrane; conventional high-resolution microscopy; immunoelectron microscopy