Phospholipase Cζ mRNA expression and its potency during spermatogenesis for activation of quail oocyte as a sperm factor

Shusei Mizushima, Soichi Takagi, Tamao Ono, Yusuke Atsumi, Akira Tsukada, Noboru Saito, Kiyoshi Shimada

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

This study was conducted to investigate the role of a sperm-borne compound in oocyte activation in special reference to the time when oocyte activation is required by testicular cells during spermatogenesis in quail. First, effects of a microinjection of quail sperm extract (SE) and quail phospholipase Cζ (PLCζ) cRNA into quail oocytes were assessed by observation of pronuclear formation and cytoplasmic segmentation, respectively. Secondly, the effects of a microinjection of round spermatids with or without PLCζ cRNA into quail oocytes were studied by observation of development. When the oocytes were injected with SE at 0.13 mg protein/ml, both pronuclear formation and cytoplasmic segmentation were optimally induced. However, pronuclear formation was blocked when SE was pretreated with heat or when the oocyte was pretreated with BAPTA (a Ca2+ chelator) before SE injection. On the other hand, when the oocytes were injected with PLCζ cRNA at 60 mg/ml, not only pronuclear formation but also cytoplasmic segmentation were optimally induced. However, PLCζ cRNA-induced pronuclear formation was blocked by pretreatment with cycloheximide (an inhibitor of protein synthesis) or with BAPTA. Most interestingly, round spermatids alone cannot induce blastodermal development but microinjection of a round spermatid with PLCζ cRNA can induce development. In addition, RT-PCR revealed that PLCζ mRNA is expressed in elongated spermatids and testicular sperm but not in round spermatids. It is concluded that PLCζ is a functional sperm factor for oocyte activation to initiate resumption of meiotic division in quail and its potency is acquired after elongated spermatid formation during the spermatogenesis.

Original languageEnglish
Pages (from-to)1200-1207
Number of pages8
JournalMolecular Reproduction and Development
Volume76
Issue number12
DOIs
Publication statusPublished - Dec 1 2009
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

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