Molecular characterization of marbled eel (Anguilla marmorata) gonadotropin subunits and their mRNA expression profiles during artificially induced gonadal development
Molecular characterization of marbled eel (Anguilla marmorata) gonadotropin subunits and their mRNA expression profiles during artificially induced gonadal development
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DOI：
10.1016/j.ygcen.2009.02.012
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发表时间：
2009-06-01
影响因子：
2.7
通讯作者：
Lin, Hao Ran
中科院分区：
文献类型：
--
作者：
Huang, Hai;Zhang, Yong;Lin, Hao Ran
从大理石纹鳗鱼中分离到3个促性腺激素亚基的cDNA序列，即共同糖蛋白α亚基（GTH α）、FSH β和LH β亚基。GTH α cDNA编码116个氨基酸，其中信号肽24个氨基酸，成熟肽92个氨基酸。FSH β亚基由127个氨基酸组成，具有22个氨基酸的信号肽和105个氨基酸的成熟肽，而LH β亚基由140个氨基酸组成，具有24个氨基酸的信号肽和116个氨基酸的成熟肽。大理石鳗鱼GTH α，FSH β和LH β与其他鱼类的推导的氨基酸序列的比较显示，在半胱氨酸残基的数量和潜在的N-连接的糖基化位点的高度保守。RT-PCR结果显示，GTH α、FSH β和LH β的mRNA不仅在垂体中有表达，而且在卵巢和睾丸中也有表达。实时荧光定量PCR分析表明，在人工诱导性腺发育过程中，雌、雄鳗垂体中GTH α和LH β的转录水平逐渐升高，分别在卵黄发生后期和精子形成期达到峰值。雌鳗垂体中FSH β mRNA在卵黄发生前期、卵黄发生早期和卵黄发生中期维持较高水平，在卵黄发生晚期和移行核期急剧下降。雄鳗垂体中FSH β mRNA的表达水平在精子发生早期高于精子发生晚期和排精期。这些结果表明，FSH将在启动和维持的性腺生长和配子发生的控制，而LH将参与最终的性腺成熟和精子/排卵的热带花鳗鲡。(C)2009 Elsevier Inc. All rights reserved.
Three cDNA sequences encoding the gonadotropin subunits, common glycoprotein alpha subunit (GTH alpha), FSH beta and LH beta subunits were isolated from marbled eel. The cDNA of GTH alpha encodes 116 amino acids with a signal peptide of 24 amino acids and a mature peptide of 92 amino acids. The FSH beta subunit consists of 127 amino acids with a 22 amino acid signal peptide and a 105 amino acid mature peptide, while the LH beta subunit consists of 140 amino acids with a 24 amino acid signal peptide and a 116 amino acid mature peptide. Comparison of the deduced amino acid sequences of marbled eel GTH alpha, FSH beta, and LH beta with that of other fishes shows a high degree of conservation in the number of cysteine residues and potential N-linked glycosylation sites. The mRNA of GTH alpha, FSH beta and LH beta were not only detected in pituitary, but also in ovary and testes by RT-PCR. Quantitative realtime PCR analysis revealed that the GTH alpha and LH beta transcriptional levels in pituitaries of female and male eels gradually increased during the artificially inducing gonadal development, and peaked at late vitellogenic stage and spermiation stage, respectively. FSH beta mRNA in the pituitaries of female eels maintained a high level at previtellogenic stage, early vitellogenic stage as well as mid-vitellogenic stage but declined sharply at late vitellogenic stage and migratory nucleus stage. In male eels, the mRNA levels of FSH beta in the pituitaries were higher at early spermatogenesis stage than at both late spermatogenesis stage and spermiation stage. These results suggested that FSH would be in control of initiation and maintenance of gonadal growth and gametogenesis, whereas LH would be involved in the final gonadal maturation and spermiation/ovulation in the tropic eel Anguilla marmorata. (C) 2009 Elsevier Inc. All rights reserved.