Measurement of intracellular IP3 during Ca2+ oscillations in mouse eggs with GFP-based FRET probe.

Abstract:

:Intracellular Ca2+ oscillations in fertilized mammalian eggs, the key signal that stimulates egg activation and early embryonic development, are regulated by inositol 1,4,5-trisphosphate (IP3) signaling pathway. We investigated temporal changes in intracellular IP3 concentration ([IP3]i) in mouse eggs, using a fluorescent probe based on fluorescence resonance energy transfer between two green fluorescent protein variants, during Ca2+ oscillations induced by fertilization or expression of phospholipase Czeta (PLCzeta), an egg-activating sperm factor candidate. Fluorescence measurements suggested the elevation of [IP3]i in fertilized eggs, and the enhancement of PLCzeta-mediated IP3 production by cytoplasmic Ca2+ was observed during Ca2+ oscillations or in response to CaCl2 microinjection. The results supported the view that PLCzeta is the sperm factor to stimulate IP3 pathway, and suggested that high Ca2+ sensitivity of PLCzeta activity and positive feedback from released Ca2+ are important for triggering and maintaining Ca2+ oscillations.

authors

Shirakawa H,Ito M,Sato M,Umezawa Y,Miyazaki S

doi

10.1016/j.bbrc.2006.04.133

subject

Has Abstract

pub_date

2006-06-30 00:00:00

pages

781-8

issue

2

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(06)00970-3

journal_volume

345

pub_type

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