These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: PI-PLC releases a 25-40 kDa protein cluster from the hamster oolemma and affects the sperm penetration assay.
    Author: Coonrod S, Naaby-Hansen S, Shetty J, Herr J.
    Journal: Mol Hum Reprod; 1999 Nov; 5(11):1027-33. PubMed ID: 10541564.
    Abstract:
    The effects of phosphatidylinositol-specific phospholipase C (PI-PLC) on human sperm-hamster oocyte interaction were investigated to determine whether PI-PLC cleavable glycosylphosphatidyinositol (GPI)-anchored proteins are involved in sperm-egg binding and fusion. Two-dimensional electrophoresis was then utilized to visualize proteins released from hamster oocytes following PI-PLC treatment. For the binding and fusion assay, either spermatozoa or eggs were treated with 1 IU/ml PI-PLC for 30 min and washed prior to gamete co-incubation. Treatment of human spermatozoa with PI-PLC significantly (P </= 0.05) enhanced sperm-egg binding while having no effect on sperm-egg fusion. Treatment of zona-free hamster oocytes with PI-PLC blocked sperm-egg binding and fusion. In order to identify the oolemmal GPI-anchored proteins involved in fertilization, egg surface proteins were labelled with sulpho-NHS biotin and either mock treated or treated with PI-PLC. Egg protein extracts and egg supernatant proteins from each group were then analysed by two-dimensional gel electrophoresis followed by avidin blotting. Comparison of blots demonstrated that a predominant biotinylated 25-40 kDa protein cluster (pI 5-6) apparent in the mock treated egg extract blot was absent in the PI-PLC treated egg extract blot. A protein cluster of identical molecular weight and isoelectric point as the predominant 25-40 kDa protein cluster was observed in the PI-PLC supernatant blot while no proteins could be seen in the control supernatant blot. These results demonstrate that treatment of hamster oocytes with PI-PLC inhibits sperm-egg interaction and releases a 25-40 kDa protein cluster (pI 5-6) from the oolemma. It is likely that this released protein cluster represents an oolemmal GPI-linked surface protein(s) which is involved in human sperm-hamster egg interaction.
    [Abstract] [Full Text] [Related] [New Search]