Suzuki Osamu, Koura Minako, Noguchi Yoko, Takano Kaoru, Uchio-Yamada Kozue, Matsuda Junichiro
Laboratory of Experimental Animal Models, National Institute of Biomedical Innovation, Saito-Asagi 7-6-8, Ibaraki, Osaka 567-0085, Japan.
Gen Comp Endocrinol. 2007 Feb;150(3):514-9. doi: 10.1016/j.ygcen.2006.09.012. Epub 2006 Nov 13.
To examine the molecular basis for efficient induction of superovulation in the rabbit, we determined the cDNA sequences of the luteinizing hormone beta-subunit (LHB) from Japanese White (JW), New Zealand White (NZW), and Dutch-Belted (Dutch) rabbits, and we compared these LHB sequences with those of other mammals. Using 5'- and 3'-rapid amplification of cDNA ends (RACE) with pituitary cDNA libraries, we found that the LHB cDNAs of all three breeds are the same length (523 bp from the 5'-end to the polyA site) and have putative AATAAA polyadenylation signal sequences at nucleotides 504 to 509. Northern blot analysis indicated that the approximately 600-nt mRNA encoding JW LHB is slightly longer than the LHB mRNAs of the other two breeds. The NZW and Dutch rabbit LHB coding sequences are 426 bp long, and their G+C contents are higher (>73%) than those of other mammalian LHBs (<70%). The predicted 141-amino-acid sequences of the JW and NZW LHB proteins are identical, and the Dutch LHB and JW/NZW sequences differ at only two residues. The exon-intron configuration of the NZW LHB gene (three exons and two introns) is similar to that of other mammalian LHB genes, and the sequences of NZW rabbit and other mammalian LHB promoter regions are highly conserved. Phylogenetic analysis of the deduced amino acid sequences of the three rabbit LHB proteins indicated that the rabbit occupies a phylogenetic position between rodents and domestic animals, and is far from humans. The results suggest that LH prepared from rodents or domestic animals, if available, would be a better inducer for superovulation in rabbits than human LH/CG.
为了研究家兔高效诱导超排卵的分子基础,我们测定了日本白兔(JW)、新西兰白兔(NZW)和荷兰带兔(Dutch)促黄体生成素β亚基(LHB)的cDNA序列,并将这些LHB序列与其他哺乳动物的序列进行了比较。利用垂体cDNA文库进行5'和3' cDNA末端快速扩增(RACE),我们发现所有三个品种的LHB cDNA长度相同(从5'端到多聚腺苷酸位点为523 bp),并且在核苷酸504至509处具有推定的AATAAA多聚腺苷酸化信号序列。Northern印迹分析表明,编码JW LHB的约600 nt mRNA略长于其他两个品种的LHB mRNA。NZW和荷兰兔LHB编码序列长426 bp,其G+C含量高于其他哺乳动物LHB(>73%)(<70%)。JW和NZW LHB蛋白预测的141个氨基酸序列相同,荷兰LHB与JW/NZW序列仅在两个残基处不同。NZW LHB基因的外显子-内含子结构(三个外显子和两个内含子)与其他哺乳动物LHB基因相似,NZW兔和其他哺乳动物LHB启动子区域的序列高度保守。对三种兔LHB蛋白推导氨基酸序列的系统发育分析表明,兔在系统发育上位于啮齿动物和家畜之间,与人类相距甚远。结果表明,如果有从啮齿动物或家畜制备的LH,它将比人LH/CG更适合作为家兔超排卵的诱导剂。