The dynamics of PKC-induced phosphorylation triggered by Ca2+ oscillations in mouse eggs.

Journal of Cellular Physiology
Jose Raul Gonzalez-GarciaK Swann

Abstract

Fertilization of mammalian eggs is characterized by a series of Ca(2+) oscillations triggered by a phospholipase C activity. These Ca(2+) increases and the parallel generation of diacylglycerol (DAG) stimulate protein kinase C (PKC). However, the dynamics of PKC activity have not been directly measured in living eggs. Here, we have monitored the dynamics of PKC-induced phosphorylation in mouse eggs, alongside Ca(2+) oscillations, using fluorescent C-kinase activity reporter (CKAR) probes. Ca(2+) oscillations triggered either by sperm, phospholipase C zeta (PLCζ) or Sr(2+) all caused repetitive increases in PKC-induced phosphorylation, as detected by CKAR in the cytoplasm or plasma membrane. The CKAR responses lasted for several minutes in both the cytoplasm and plasma membrane then returned to baseline values before subsequent Ca(2+) transients. High frequency oscillations caused by PLCζ led to an integration of PKC-induced phosphorylation. The conventional PKC inhibitor, Gö6976, could inhibit CKAR increases in response to thapsigargin or ionomycin, but not the repetitive responses seen at fertilization. Repetitive increases in PKCδ activity were also detected during Ca(2+) oscillations using an isoform-specific δCKAR. However,...Continue Reading

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Citations

May 31, 2014·BioMed Research International·Md Saidur RahmanMyung-Geol Pang
Sep 9, 2015·Human Reproduction Update·Marc YesteKevin Coward
Sep 9, 2016·Cellular and Molecular Life Sciences : CMLS·Jacinta H MartinBrett Nixon
Feb 12, 2017·Cell Research·Guy A Rutter
Nov 11, 2017·Reproduction : the Official Journal of the Society for the Study of Fertility·Minerva Ferrer-BuitragoBjörn Heindryckx
Jul 24, 2015·Molecular Reproduction and Development·Wei MaMaria M Viveiros
Oct 12, 2019·Experimental and Therapeutic Medicine·Yanchun LiuBingzhi Yu

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Methods Mentioned

BETA
PMA
fluorescence resonance energy transfer
FRET
myristoylation
for
by

Software Mentioned

Vivo
Image J
SigmaPlot
In

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