title: Munc18-1 controls SNARE protein complex assembly during human sperm acrosomal exocytosis.
abstract: The spermatozoon is a very specialized cell capable of carrying out a limited set of functions with high efficiency. Sperm are then excellent model cells to dissect fundamental processes such as regulated exocytosis. The secretion of the single dense-core granule of mammalian spermatozoa relies on the same highly conserved molecules and goes through the same stages as exocytosis in other types of cells. In this study, we describe the presence of Munc18-1 in human sperm and show that this protein has an essential role in acrosomal exocytosis. We observed that inactivation of endogenous Munc18-1 with a specific antibody precluded the stabilization of trans-SNARE complexes and inhibited acrosomal exocytosis. Addition of recombinant Munc18-1 blocked secretion by sequestering monomeric syntaxin, an effect that was rescued by α-soluble NSF attachment protein. By electron microscopy, we observed that both the anti-Munc18-1 antibody and recombinant Munc18-1 inhibited the docking of the acrosome to the plasma membrane. In conclusion, our results indicate that Munc18-1 plays a key role in the dynamics of trans-SNARE complex assembly and/or stabilization, a process that is necessary for the docking of the outer acrosomal membrane to the plasma membrane and subsequent fusion pore opening.
authors.affiliation: Laboratorio de Biología Celular y Molecular, Instituto de Histología y Embriología-Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, 5500 Mendoza, Argentina.
authors.forename: Facundo
authors.initials: F
authors.lastname: Rodríguez
authors.forename: M Natalia
authors.initials: MN
authors.lastname: Zanetti
authors.forename: Luis S
authors.initials: LS
authors.lastname: Mayorga
authors.forename: Claudia N
authors.initials: CN
authors.lastname: Tomes
mesh_terms: D000177:Acrosome / Q000378:metabolism* / Q000648:ultrastructure; D020101:Acrosome Reaction / Q000502:physiology; D000906:Antibodies / Q000737:chemistry; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D005089:Exocytosis / Q000502:physiology*; D006801:Humans; D008297:Male; D051938:Munc18 Proteins / Q000235:genetics / Q000378:metabolism*; D055550:Protein Stability; D050600:SNARE Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000906:Antibodies; D051938:Munc18 Proteins; D050600:SNARE Proteins