Kim Dae-Jung, Cho Yong Chul, Sohn Young Chang
National Fisheries Research and Development Institute, Gijang-Gun, Busan 619-902, Republic of Korea.
Gen Comp Endocrinol. 2005 May 1;141(3):282-90. doi: 10.1016/j.ygcen.2005.01.001.
In an attempt to understand the reproductive regulation in viviparous teleosts, gonadotropin (GTH) subunit cDNAs were characterized and the expression levels of GTH subunit mRNAs in the pituitary glands of the rockfish, Sebastes schlegeli (Scorpaeniformes, Scorpaenidae), were examined by Northern blot analysis. The complete sequences of rockfish GTH subunits (GTHalpha, FSHbeta, and LHbeta) were determined by rapid amplification of cDNA ends (RACE) and nucleotide sequencing. Based on the RACE analysis, the cDNAs of GTHalpha, FSHbeta, and LHbeta consisted of 655, 540, and 529 nucleotides encoding peptides of 132, 130, and 143 amino acids, respectively. The mature peptides of rockfish FSHbeta, LHbeta, and common GTHalpha showed high sequence identities (FSHbeta, 58-62%; LHbeta, 86-94%; and GTHalpha, 87-88%) to those of other perciforme (e.g., orange spotted grouper, red seabream, stripped bass, and sea bass). In a sequence alignment of the mature peptides, rockfish FSHbeta exhibited a unique feature, the lack of a conserved N-glycosylation site. This is in contrast to both LHbeta and GTHalpha which contain one and two N-glycosylation sites, respectively, and is consistent with those of other teleosts. The mRNA levels of the GTHalpha subunit increased at the vitellogenic stage and remained steady from ovulation to post-parturition. FSHbeta mRNA levels increased abruptly during the vitellogenic stages and then decreased during ovulation, embryonic development, and post-parturition. LHbeta mRNA levels were observed to rapidly increase during the vitellogenic stage, reached its highest levels during ovulation and was then followed by a decrease at post-parturition. These results suggest that in the female rockfish FSH and LH syntheses are predominant during vitellogenesis and oocyte maturation, respectively.
为了了解胎生硬骨鱼的生殖调控机制,对促性腺激素(GTH)亚基cDNA进行了表征,并通过Northern印迹分析检测了条石鲷(鲈形目,鲉科)垂体中GTH亚基mRNA的表达水平。通过cDNA末端快速扩增(RACE)和核苷酸测序确定了条石鲷GTH亚基(GTHα、FSHβ和LHβ)的完整序列。基于RACE分析,GTHα、FSHβ和LHβ的cDNA分别由655、540和529个核苷酸组成,分别编码132、130和143个氨基酸的肽段。条石鲷FSHβ、LHβ和常见GTHα的成熟肽与其他鲈形目鱼类(如点带石斑鱼、真鲷、条纹鲈和海鲈)的成熟肽具有较高的序列同一性(FSHβ为58 - 62%;LHβ为86 - 94%;GTHα为87 - 88%)。在成熟肽的序列比对中,条石鲷FSHβ表现出一个独特的特征,即缺乏保守的N - 糖基化位点。这与LHβ和GTHα相反,LHβ和GTHα分别含有一个和两个N - 糖基化位点,并且与其他硬骨鱼一致。GTHα亚基的mRNA水平在卵黄生成期升高,从排卵到产后保持稳定。FSHβ mRNA水平在卵黄生成期急剧升高,然后在排卵、胚胎发育和产后下降。观察到LHβ mRNA水平在卵黄生成期迅速升高,在排卵时达到最高水平,然后在产后下降。这些结果表明,在雌性条石鲷中,FSH和LH的合成分别在卵黄生成和卵母细胞成熟过程中占主导地位。