Seeing is believing: Current methods to observe sperm acrosomal exocytosis in real time

Mol Reprod Dev. 2020 Dec;87(12):1188-1198. doi: 10.1002/mrd.23431. Epub 2020 Oct 28.

Abstract

Acrosomal exocytosis (AR) is a critical process that sperm need to undergo to fertilize an egg. The evaluation of the presence or absence of the acrosome is usually performed by using lectins or dyes in fixed cells. With this approach, it is neither possible to monitor the dynamic process of exocytosis and related molecular events while discriminating between live and dead cells, nor to evaluate the acrosomal status while sperm reside in the female reproductive tract. However, over the last two decades, several new methodologies have been used to assess the occurrence of AR in living cells allowing different groups to obtain information that was not possible in the past. These techniques have revolutionized the whole study of this process. This review summarizes current methods available to analyze AR in living cells as well as the important information that emerged from studies using these approaches.

Keywords: EGFP-sperm; FM4-64; acrosome reaction; flow cytometry; live imaging; single cell.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Acrosome / metabolism*
  • Acrosome Reaction / physiology*
  • Actin Cytoskeleton / metabolism
  • Animals
  • Calcium / metabolism
  • Exocytosis / physiology*
  • Female
  • Fertilization in Vitro / methods*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Male
  • Mice
  • Mice, Transgenic
  • Sperm Capacitation / physiology*
  • Zona Pellucida / metabolism

Substances

  • Green Fluorescent Proteins
  • Calcium