Molecular cloning, testicular postnatal expression, and oocyte-activating potential of porcine phospholipase Czeta.
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.
Yoneda
Akihiro
A
Laboratory of Animal Breeding and Reproduction, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
Kashima
Masashi
M
Yoshida
Shigeki
S
Terada
Kei
K
Nakagawa
Shoma
S
Sakamoto
Akiko
A
Hayakawa
Koji
K
Suzuki
Keita
K
Ueda
Junji
J
Watanabe
Tomomasa
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000032:analysis / Q000378:metabolism; D020013:Calcium Signaling; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005260:Female; D005307:Fertilization in Vitro; D015870:Gene Expression; D008297:Male; D051379:Mice; D008845:Microinjections; D008969:Molecular Sequence Data; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D009866:Oogenesis / Q000187:drug effects / Q000502:physiology*; D018075:RNA, Complementary / Q000494:pharmacology; D016415:Sequence Alignment; D013091:Spermatogenesis; D013552:Swine / Q000378:metabolism*; D013737:Testis / Q000201:enzymology* / Q000254:growth & development; D010738:Type C Phospholipases / Q000235:genetics / Q000502:physiology*
D018075:RNA, Complementary; D004247:DNA; D010738:Type C Phospholipases; D002118:Calcium