Activation of follicle cell surface phospholipase by tyrosine kinase dependent pathway is an essential event in Ascidian fertilization.

Abstract:

:Eggs of Ascidia ceratodes and Phallusia mammillata block polyspermy by releasing a phosphatidylinositol-linked glycosidase from the follicle cell and egg surface that binds to and blocks all unoccupied sperm binding sites on the vitelline coat. Release of this glycosidase is thought to be under the control of a membrane-bound phospholipase. To elucidate the mechanism of phospholipase activation, intact eggs and isolated follicle cells are activated by either sperm or the tyrosine kinase activator 9, 10-dimethyl-1,2-benzanthracene (DMBA). Both treatments caused release of comparable quantities of glycosidase activity, the earliest event following fertilization. A corresponding increase in phospholipase activity accompanied this glycosidase release. The tyrosine kinase inhibitor genistein blocked release by DMBA at concentrations as low as 1 microM, but had no effect on sperm-induced release even when used up to 100 microM. Tyrphostin A23, another tyrosine kinase inhibitor, when used at 200 microM blocked glycosidase release and decreased phospholipase activity following both DMBA activation and fertilization. Western blot analysis probing for phosphotyrosine content of disrupted intact eggs with their follicle cells revealed the absence of a band in tyrphostin-treated eggs corresponding to a 40 kDa protein that was present in both unfertilized and fertilized egg samples. Based on these results, we propose that phosphorylation of specific tyrosine residues is necessary for phospholipase activation and is sufficient to trigger subsequent glycosidase release.

journal_name

Mol Reprod Dev

authors

Robert LK,Lucio-Gough LM,Goode CA,Mckinney K,Lambert CC

doi

10.1002/(SICI)1098-2795(199909)54:1<69::AID-MRD10>

keywords:

subject

Has Abstract

pub_date

1999-09-01 00:00:00

pages

69-75

issue

1

eissn

1040-452X

issn

1098-2795

pii

10.1002/(SICI)1098-2795(199909)54:1<69::AID-MRD10>

journal_volume

54

pub_type

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