Nomikos, Michail, Sanders, Jessica Rose, Theodoridou, Maria, Kashir, Junaid, Matthews, Emily, Nounesis, George, Lai, Francis ORCID: https://orcid.org/0000-0003-2852-8547 and Swann, Karl ORCID: https://orcid.org/0000-0002-4355-1449 2014. Sperm-specific post-acrosomal WW-domain binding protein (PAWP) does not cause Ca2+ release in mouse oocytes. Molecular Human Reproduction 20 (10) , pp. 938-947. 10.1093/molehr/gau056 |
Abstract
Mature mammalian oocytes undergo a prolonged series of cytoplasmic calcium (Ca2+) oscillations at fertilization that are the cause of oocyte activation. The Ca2+ oscillations in mammalian oocytes are driven via inositol 1,4,5-trisphosphate (IP3) generation. Microinjection of the sperm-derived phospholipase C-zeta (PLCζ), which generates IP3, causes the same pattern of Ca2+ oscillations as observed at mammalian fertilization and it is thought to be the physiological agent that triggers oocyte activation. However, another sperm-specific protein, ‘post-acrosomal WW-domain binding protein’ (PAWP), has also been reported to elicit activation when injected into mammalian oocytes, and to produce a Ca2+ increase in frog oocytes. Here we have investigated whether PAWP can induce fertilization-like Ca2+ oscillations in mouse oocytes. Recombinant mouse PAWP protein was found to be unable to hydrolyse phosphatidylinositol 4,5-bisphosphate in vitro and did not cause any detectable Ca2+ release when microinjected into mouse oocytes. Microinjection with cRNA encoding either the untagged PAWP, or yellow fluorescent protein (YFP)-PAWP, or luciferase-PAWP fusion proteins all failed to trigger Ca2+ increases in mouse oocytes. The lack of response in mouse oocytes was despite PAWP being robustly expressed at similar or higher concentrations than PLCζ, which successfully initiated Ca2+ oscillations in every parallel control experiment. These data suggest that sperm-derived PAWP is not involved in triggering Ca2+ oscillations at fertilization in mammalian oocytes.
Item Type: | Article |
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Date Type: | Publication |
Status: | Published |
Schools: | Biosciences Medicine |
Subjects: | R Medicine > R Medicine (General) |
Publisher: | Oxford University Press |
ISSN: | 1360-9947 |
Date of Acceptance: | 18 July 2014 |
Last Modified: | 16 Dec 2023 15:40 |
URI: | https://orca.cardiff.ac.uk/id/eprint/78531 |
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