-
View Affiliations Hide Affiliations
- Publisher: Hamad bin Khalifa University Press (HBKU Press)
- Source: Qatar Foundation Annual Research Conference Proceedings, Qatar Foundation Annual Research Conference Proceedings Volume 2014 Issue 1, Nov 2014, Volume 2014, HBPP0176
oa Role Of Stim1 And Orai1 In Mammalian Oocyte Activation
Abstract
Upon fertilization, the mammalian oocyte undergoes a series of Ca2+ oscillations, which results in its activation and initiation of embryo development. It is believed that these periodic and extended Ca2+ responses are a consequence of intracellular Ca2+ release coupled to Ca2+ influx across the plasma membrane due to the activation of a process known as Store Operated Calcium Entry (SOCE). The underlying mechanism of SOCE is known to be controlled by Stim-Orai channels, where IP3-mediated emptying of ER calcium stores results in the aggregation of Stim1 molecules followed by interaction with Orai1 channels on the plasma membrane and activation of Ca2+ entry. Stim1 was shown to redistribute in the form of patches after mouse oocyte fertilization in a manner similar to its redistribution after pharmacological Ca2+ store depletion, while down-regulation of Orai1 expression in pig oocytes inhibited oscillations induced by fertilization. We have further shown that overexpression of Stim1 and Orai1 in mouse oocytes disrupts Ca2+ dynamics and inhibits egg activation. These data support a role for SOCE in Ca2+ signaling during fertilization. To further investigate the importance of Stim1 and Orai1 proteins in regulating events of egg activation, we are generating oocyte-specific Stim1 and Orai1 knockout mice by crossing Stim1-flox and Orai1-flox mice with ZP3-Cre transgenic mice. ZP3 is expressed only in oocytes leading to Cre expression only in this cell type and as such an oocyte-specific knockout. Studying rates of fertilization and Ca2+ entry in these knockout oocytes will provide insight into the role of these two proteins in the initiation and persistence of Ca2+oscillations after fertilization, and will help us determine whether there is an absolute requirement for Stim1 and/or Orai1 in regulating Ca2+ responses in early stages of fertilization.