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Calcium oscillations in fertilized pig oocytes are associated with repetitive interactions between STIM1 and ORAI1

Lu Zhang1, Chi-Hong Chao2, Laurie A Jaeger3

  • 1Department of Animal Sciences, Purdue University, West Lafayette, Indiana, USA.

Biology of Reproduction
|January 25, 2018
PubMed

Insights

Stromal interaction molecule 1 (STIM1) and ORAI1 proteins mediate calcium (Ca2+) entry in pig oocytes, sustaining fertilization-induced Ca2+ oscillations. This store-operated Ca2+ entry (SOCE) mechanism is crucial for oocyte activation and development.

Area of Science:

  • Cell Biology
  • Reproductive Biology
  • Ion Channel Physiology

Background:

  • Fertilized oocytes exhibit characteristic calcium (Ca2+) oscillations, essential for triggering developmental processes.
  • The precise molecular mechanisms governing Ca2+ influx in pig oocytes remain incompletely understood.
  • Stromal interaction molecule 1 (STIM1) and ORAI1 are key components of store-operated Ca2+ entry (SOCE) pathways in many cell types.

Purpose of the Study:

  • To investigate the role of STIM1 and ORAI1 in mediating Ca2+ entry that sustains Ca2+ oscillations in fertilized pig oocytes.
  • To elucidate the spatiotemporal interaction between STIM1 and ORAI1 during store-operated Ca2+ entry (SOCE) in pig oocytes.
  • To determine the functional significance of STIM1-ORAI1 interaction in regulating Ca2+ dynamics during fertilization.

Main Methods:

  • Expression of fluorescently tagged STIM1 and ORAI1 proteins in pig oocytes.
  • Induction of Ca2+ store depletion using cyclopiazonic acid (CPA).
  • Monitoring of cytosolic Ca2+ concentration and investigation of STIM1-ORAI1 interaction using fluorescence resonance energy transfer (FRET).
  • Pharmacological inhibition of STIM1 puncta formation and SOCE.

Main Results:

  • Store depletion induced STIM1-ORAI1 interaction, evidenced by increased FRET signals.
  • STIM1 puncta formation was observed upon store depletion and was inhibited by ML-9, which also blocked SOCE and fertilization-induced Ca2+ oscillations.
  • Overexpression of STIM1 or co-expression of STIM1 and ORAI1 led to high-frequency Ca2+ oscillations upon fertilization, which were sensitive to SOCE inhibitors.
  • Cyclic FRET increases indicated repetitive STIM1-ORAI1 interactions during fertilization.

Conclusions:

  • The study confirms the involvement of store-operated Ca2+ entry (SOCE) mediated by STIM1 and ORAI1 in maintaining repetitive Ca2+ transients during pig oocyte fertilization.
  • STIM1 and ORAI1 interaction is a critical event for sustaining the Ca2+ oscillations necessary for oocyte activation.
  • These findings provide molecular insights into the Ca2+ signaling machinery regulating mammalian oocyte fertilization.

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