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Several Phospholipase C (PLC) isof orms have been found in male and female mammalian gametes and splicing isoforms of PLCδ4 are predominantly expressed in testis. To understand a physiological function of PLCδ4, we generated <I>PLCδ4</I> gene-deficient mice. PLCδ4 gene-disrupted male mice either produced few small litters or were sterile. In vivo and in vitro fertilization studies showed that PLCδ4 has a functional role in sperm at the early step of fertilization. We also observed that the calcium transients in eggs associated with fertilization were absent or delayed when we used PLCδ4<SUP>-/-</SUP> sperm. PLCδ4<SUP>-/-</SUP> sperm were unable to initiate the acrosome reaction, an exocytotic event required for fertilization and induced by interaction with the egg coat, the zona pellucida (ZP) . Next we have monitored Ca<SUP>2+</SUP> responses in single sperm, and we found that the [Ca<SUP>2+</SUP>] <SUB>i</SUB> increase in response to ZP is absent in PLCδ4<SUP>-/-</SUP> sperm. Progesterone, another physiological inducer of the acrosome reaction, failed to induce sustained [Ca<SUP>2+</SUP>] <SUB>i</SUB> increases in PLCδ<SUP>-/-</SUP> sperm and consequently the acrosome reaction was partially inhibited. Calcium imaging studies revealed that the [Ca<SUP>2+</SUP>] <SUB>i</SUB> increases induced by exposure to ZP and progesterone started at different sites within the sperm head, indicating that these agonists induce the acrosome reaction via different Ca<SUP>2+</SUP> mechanisms. Furthermore, storeoperated channel (SOC) activity was severely impaired in PLCδ4<SUP>-/-</SUP> sperm. These results support that PLCδ4 is an essential protein for calcium mobilization in sperm that drives acrosome reactions.
- 公益社団法人 日本実験動物学会の論文
公益社団法人 日本実験動物学会 | 論文
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