Premium
Identification of phospholipase C (PLC ) and its possible roles in egg activation in Chaetopterus eggs
Author(s) -
Yin Xunqin,
Eckberg William R,
Eckberg William R
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.lb99-d
Subject(s) - oocyte activation , biology , phospholipase c , protein kinase c , second messenger system , microbiology and biotechnology , signal transduction , complementary dna , gene isoform , biochemistry , oocyte , gene , embryo
Phospholipase C (PLC) is a critical enzyme that is involved in generation of the second messenger inositol‐1,4,5‐trisphosphate (IP 3 ) for the calcium release that brings about egg activation in vertebrates and invertebrates. In the present study, we for the first time identified a novel PLC isoform, termed cp‐PLC γ , in Chaetopterus eggs by reverse transcription/degenerate oligonucleotide primered polymerase chain reaction (RT/DOP‐PCR) and rapid amplification of cDNA end polymerase chain reaction (RACE PCR) strategies. Cp‐PLC γ is expressed as a 150kDa protein in unfertilized and fertilized Chaetopterus eggs. The general PLC antagonist U‐73122 blocked egg activation in response to fertilization. However, the inactive analog, U‐73343, had no effect on early embryonic development. We further determined that protein tyrosine kinase (PTK) antagonists, genistein and tyrophostein B42, inhibit both normal and parthenogenetic egg activation in a dose‐dependent manner. These data indicates that PLC activity from the eggs is required for egg activation and that PLC γ may play an important role by PTK‐PLC γ signal cascade. This research was supported by the NSF HRD‐0401697