Activation of mouse oocytes requires multiple sperm factors but not sperm PLCgamma1.
A sperm cytosolic factor is responsible for oocyte activation at fertilization in mammals. The molecular identity of this factor is not yet known, although a sperm phospholipase Cgamma (PLCgamma) is a potential candidate. In this study, cation-exchange chromatography with a Heparin column was used for the fractionation of porcine sperm cytosolic extracts. Oocyte activation potential of the resulting fractions was tested and active fractions were subjected to Western blot analysis using antibodies specific to PLCgamma1. PLCgamma1 was detected in fractions other than those supporting oocyte activation (Ca(2+)-release and pronuclear formation). The active Heparin fraction was then purified on a Mono Q anion-exchange column. One of the resulting fractions still contained Ca(2+)-releasing activity, but pronuclear formation did not occur. We conclude that sperm PLCgamma1 is not involved in oocyte activation and that Ca(2+)-release and pronuclear formation requires multiple factors from sperm cytosol.
Heyers
S
S
Department of Endocrinology and Reproductive Medicine, Universitäts-Frauenklinik Bonn, Sigmund-Freud-Str. 25, 53105, Bonn, Germany.
Sousa
M
M
Cangir
O
O
Schmoll
F
F
Schellander
K
K
van der Ven
H
H
Montag
M
M
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002467:Cell Nucleus / Q000502:physiology; D003600:Cytosol / Q000502:physiology; D005260:Female; D005306:Fertilization / Q000502:physiology*; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000502:physiology*; D008297:Male; D051379:Mice; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D051966:Phospholipase C gamma; D013094:Spermatozoa / Q000502:physiology*; D013552:Swine; D010738:Type C Phospholipases / Q000502:physiology*
D007527:Isoenzymes; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D002118:Calcium