Synaptotagmin VI participates in the acrosome reaction of human spermatozoa

Marcela Michaut, Gerardo De Blas, Claudia N. Tomes, Roberto Yunes, Mitsunori Fukuda, Luis S. Mayorga

Research output: Contribution to journalArticlepeer-review

64 Citations (Scopus)

Abstract

Acrosomal exocytosis is a calcium-dependent secretion event causing the release of the acrosomal contents and the loss of the membranes surrounding the acrosome. The synaptotagmins are a family of calcium-binding proteins that participate in the exocytosis of synaptic vesicles. The ubiquitous synaptotagmin VI isoform was found in human sperm cells by Western blot analysis. Immunocytochemistry at the optical and electron microscopy level's localized the protein to the outer acrosomal membrane. Calcium-triggered acrosomal exocytosis in permeabilized sperm cells was abrogated by a specific anti-synaptotagmin VI antibody, indicating that the protein is required for the process. Moreover, a recombinant fusion protein between glutathione S-transferase and the two calcium and phospholipid binding domains of synaptotagmin VI completely inhibited calcium-triggered exocytosis. Interestingly, phorbol ester-dependent in vitro phosphorylation of this recombinant protein abolished its inhibitory effect. We previously showed that, in permeabilized spermatozoa, addition of active Rab3A triggers acrosomal exocytosis at very low calcium concentration. Rab3A-promoted exocytosis was inhibited by the cytosolic domain of synaptotagmin VI and by the anti-synaptotagmin VI antibody, indicating that synaptotagmin is also necessary for Rab-mediated acrosomal content release. In conclusion, the results strongly indicate that synaptotagmin VI is a key component of the secretory machinery involved in acrosomal exocytosis.

Original languageEnglish
Pages (from-to)521-529
Number of pages9
JournalDevelopmental Biology
Volume235
Issue number2
DOIs
Publication statusPublished - 2001 Jul 15
Externally publishedYes

Keywords

  • Acrosome reaction
  • Human spermatozoa
  • Membrane fusion
  • Protein kinase C
  • Regulated exocytosis
  • Streptolysin O
  • Synaptotagmin VI
  • Synaptotagmin phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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