Molecular cloning and hormonal control in the ovary of connexin 31.5 mRNA and correlation with the appearance of oocyte maturational competence in red seabream.
Gap junctions are aggregates of intercellular channels, composed of the protein connexin (Cx), between adjacent cells. This study examined whether, in the ovary of the red seabream Pagrus major, the connexin gene essential for the production of RNA and protein during the acquisition of oocyte maturational competence is active. Mixed primers for this reaction were designed on the basis of the high sequence homology of selected regions of known connexin genes. Polymerase-chain-reaction-amplified cDNA fragments generated by 3' and 5' rapid amplication of cDNA ends were combined to generate full-length cDNA sequences. The resulting 2400 base pair cDNA had an open reading frame encoding a polypeptide containing 275 amino acid residues (31493 Da; Cx31.5). Hydropathicity analysis of the predicted amino acid sequence indicated that red seabream Cx31.5 has four major hydrophobic regions and four major hydrophilic regions indicative of a topology similar to that of known connexins. Typical connexin consensus sequences were also observed in the first and second extracellular loops. During the acquisition of oocyte maturational competence, red seabream Cx31.5 mRNA transcription levels increased after treatment with gonadotropin-II. It is therefore proposed that expression of Cx31.5 contributes to the acquisition of oocyte maturational competence in this species.
Choi
C Y
CY
Department of Aquatic Biosciences, Tokyo University of Fisheries, Konan, Minato-ku, Tokyo 108-8477, Japan. choi_cy@hotmail.com
Takashima
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017630:Connexins / Q000737:chemistry / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006065:Gonadotropins, Pituitary / Q000494:pharmacology; D008969:Molecular Sequence Data; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development; D010053:Ovary / Q000187:drug effects / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D021541:Sea Bream / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
D017630:Connexins; D017931:DNA Primers; D018076:DNA, Complementary; D006065:Gonadotropins, Pituitary; D012333:RNA, Messenger; C418923:connexin 31.5