Sperm-borne protein, PAWP, initiates zygotic development in Xenopus laevis by eliciting intracellular calcium release.
We previously reported postacrosomal sheath WW domain binding protein (PAWP) as a candidate sperm borne, oocyte-activating factor. PAWP enters the oocyte during fertilization and induces oocyte activation events including meiotic resumption, pronuclear formation, and egg cleavage. However, in order to provide proof that PAWP is a primary initiator of zygotic development it is imperative to show that PAWP initiates intracellular calcium signaling, which is considered essential for oocyte activation. Utilizing Xenopus laevis as our model, we injected recombinant PAWP or Xenopus sperm into metaphase II-arrested oocytes and observed a significant rise in intracellular calcium levels over controls. Concurring intensities and durations of PAWP and sperm-induced calcium waves, detected by infrared two-photon laser-scanning fluorescence microscopy, were prevented by coinjection of a competitive PPGY-containing peptide derived from PAWP but not by the point-mutated form of this peptide. This study also correlates PAWP and sperm-induced calcium release with meiotic resumption in Xenopus. The similar mode of oocyte activation, and the ability of the competitive peptide in blocking both sperm- and PAWP-induced calcium release, provide evidence for the first time that sperm-anchored PAWP is a primary initiator of zygotic development.
Aarabi
Mahmoud
M
Department of Anatomy and Cell Biology, Queen's University, Kingston, Ontario, Canada K7L 3N6.
Qin
Zheng
Z
Xu
Wei
W
Mewburn
Jeffrey
J
Oko
Richard
R
D020816:Amino Acid Motifs / Q000235:genetics; D000704:Analysis of Variance; D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D016009:Chi-Square Distribution; D005260:Female; D005306:Fertilization / Q000502:physiology; D005452:Fluoresceins; D008297:Male; D008540:Meiosis; D036641:Microscopy, Fluorescence, Multiphoton; D009154:Mutation; D009865:Oocytes / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D029589:Seminal Plasma Proteins / Q000235:genetics / Q000378:metabolism*; D013094:Spermatozoa / Q000378:metabolism; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis; D015053:Zygote / Q000737:chemistry / Q000378:metabolism*
D000814:Aniline Compounds; D002352:Carrier Proteins; D005452:Fluoresceins; C503942:Oregon green 488 BAPTA-1; D011994:Recombinant Proteins; D029589:Seminal Plasma Proteins; D029867:Xenopus Proteins; D002118:Calcium