Yoneda Akihiro, Kashima Masashi, Yoshida Shigeki, Terada Kei, Nakagawa Shoma, Sakamoto Akiko, Hayakawa Koji, Suzuki Keita, Ueda Junji, Watanabe Tomomasa
Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Reproduction. 2006 Sep;132(3):393-401. doi: 10.1530/rep.1.01018.
The molecular mechanism by which sperm triggers Ca2+ oscillation, oocyte activation, and early embryonic development has not been clarified. Recently, oocyte activation has been shown to be induced by sperm-specific phospholipase Czeta (PLCzeta). The ability of PLCzeta to induce oocyte activation is highly conserved across vertebrates. In the present study, porcine PLCzeta cDNA was identified and the nucleotide sequence was determined. The expression pattern of porcine PLCzeta mRNA during the period of postnatal testicular development was shown to be similar to that of mouse PLCzeta. PLCzeta mRNA expression in the pig and mouse was detected only in the testes when the elongated spermatids had differentiated, and was detected from day 96 after birth in the pig. Histological examination of porcine testis during the period of postnatal development revealed the presence of spermatozoa from day 110 after birth. These findings suggest that the synthesis of PLCzeta mRNA starts when spermiogenesis is initiated. Microinjection of porcine PLCzeta complementary RNA into porcine oocytes demonstrated that porcine PLCzeta has the ability to trigger repetitive Ca2+ transients in porcine oocytes similar to that observed during fertilization. It was also found that porcine PLCzeta cRNA has the potential to induce oocyte activation and initiate embryonic development up to the blastocyst stage.
精子触发钙离子振荡、卵母细胞激活和早期胚胎发育的分子机制尚未阐明。最近,已证明卵母细胞激活是由精子特异性磷脂酶Czeta(PLCzeta)诱导的。PLCzeta诱导卵母细胞激活的能力在脊椎动物中高度保守。在本研究中,鉴定了猪PLCzeta cDNA并确定了其核苷酸序列。结果表明,猪PLCzeta mRNA在出生后睾丸发育期间的表达模式与小鼠PLCzeta相似。仅在伸长的精子细胞分化时,在猪和小鼠的睾丸中检测到PLCzeta mRNA表达,在猪中从出生后第96天开始检测到。对猪出生后发育期间睾丸的组织学检查显示,从出生后第110天开始存在精子。这些发现表明,PLCzeta mRNA的合成在精子发生开始时启动。将猪PLCzeta互补RNA显微注射到猪卵母细胞中表明,猪PLCzeta具有在猪卵母细胞中触发重复性钙离子瞬变的能力,类似于受精期间观察到的情况。还发现猪PLCzeta cRNA具有诱导卵母细胞激活并启动胚胎发育直至囊胚阶段的潜力。