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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels
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Single-Cell Calcium Imaging for Studying the Activation of Calcium Ion Channels

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Two-pore channel 1 and Ca

Enrique I Oliver1,2, Martina Jabloñski3, Mariano G Buffone3

  • 1Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Autónoma de México, Cuernavaca, Morelos, México.

The Journal of Physiology
|June 6, 2023
PubMed
Summary
This summary is machine-generated.

Acrosomal alkalinization, crucial for sperm function, triggers calcium release via TPC1 and CRAC channels. This study reveals key pathways controlling calcium signaling during the acrosome reaction (AR) in mouse sperm.

Keywords:
CRAC channelsCa2+ signalsTPC1 channelsacrosomal alkalinizationacrosomal reactionacrosomal vesicle

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Area of Science:

  • Reproductive Biology
  • Cell Biology
  • Biochemistry

Background:

  • The acrosome reaction (AR) is vital for mammalian fertilization, involving calcium (Ca2+) and acrosomal alkalinization.
  • The precise molecular mechanisms of Ca2+ transport during AR, particularly those linked to pH changes, remain unclear.

Purpose of the Study:

  • To investigate the pathways mediating Ca2+ signals induced by acrosomal pH (pHa) increase in mouse sperm.
  • To elucidate the roles of TPC1 and CRAC channels in regulating Ca2+ fluxes during acrosomal alkalinization.

Main Methods:

  • Single-cell Ca2+ imaging in mouse sperm.
  • Pharmacological inhibition of sperm-specific Ca2+ channels (CatSper) and two-pore channels (TPC1).
  • Utilizing the lysosomotropic agent Gly-Phe-β-naphthylamide (GPN) to assess osmotic contributions.

Main Results:

  • Mibefradil and NNC 55-0396 induced pHa increase and acrosomal Ca2+ release without damaging the acrosomal membrane.
  • TPC1 channel inhibition reduced Ca2+ influx stimulated by acrosomal alkalinization.
  • CRAC channel blockage diminished Ca2+ uptake triggered by pHa elevation.

Conclusions:

  • Acrosomal alkalinization triggers Ca2+ release and uptake through TPC1 and CRAC channels, respectively.
  • These findings clarify the role of pHa in controlling Ca2+ efflux and influx during the acrosome reaction.
  • This research enhances understanding of the molecular basis of sperm fertilization.