Cloning and sequencing of the FSH-β and LH β-subunit in the three-spined stickleback, Gasterosteus aculeatus, and effects of photoperiod and temperature on LH-β and FSH-β mRNA expression
Cloning and sequencing of the FSH-β and LH β-subunit in the three-spined stickleback, Gasterosteus aculeatus, and effects of photoperiod and temperature on LH-β and FSH-β mRNA expression
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DOI：
10.1016/j.ygcen.2003.09.003
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发表时间：
2004-01-15
影响因子：
2.7
通讯作者：
Schmitz, M
中科院分区：
文献类型：
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作者：
Hellqvist, A;Bornestaf, C;Schmitz, M
本研究的目的是表征的FSH-β和LH-β亚基的三刺鱼，在实验研究中广泛使用的鱼，并使用克隆的cDNA作为探针测量FSH-β和LH-β mRNA表达在不同的光周期和温度处理的刺鱼。从雄性棘鱼的10个垂体制备第一链cDNA，并使用基于已知硬骨鱼FSH-β和LH-β cDNA序列的高度保守区域的简并引物通过PCR扩增cDNA片段。为了获得FSH-β和LH-β的全长cDNA，进行RACE扩增。FSH-β的cDNA长度为540，编码122个氨基酸的蛋白质，LH-β的cDNA长度为568，编码148个氨基酸的蛋白质。在迄今为止发表的促性腺激素（GTH）β亚基中，与棘鱼GTH最相似的是在其他percomorph鱼类中发现的，FSH-β亚基的氨基酸相似性为46-55%，LH-β亚基的氨基酸相似性为75-77%。以克隆的cDNA为探针，分析了不同光周期和温度下刺鱼垂体LH和FSH mRNA的表达。将200只雄性分为4组，分别置于不同的光周期和温度下：光暗比（LD）16：8，20 ℃; LD 16：8，7 ℃; LD 8：16，20 ℃和LD 8：16，7 ℃。在20 ℃下，LD为16：8的条件下，棘鱼出现红色繁殖色和明显的肾脏肥大，雄激素依赖性雄性第二性征，而在其他组中没有出现。LD 8：16，20 ℃组的肾上皮高度（KEH）显著低于其他各组，这也是唯一一个精子发生活跃的组。LD 8：16 20 ℃组FSH-β和LH-β的表达均显著低于其他组。因此，当精子发生活跃时，以及当低KEH表明雄激素水平处于最低水平时，两种GTH均显示出最低表达。(C)2003年爱思唯尔公司All rights reserved.
The aim of this study was to characterize the FSH-beta and LH-beta subunit in the three-spined stickleback, a fish used extensively in experimental studies, and to use the cloned cDNAs as probes for measuring FSH-beta and LH-beta mRNA expression in sticklebacks treated with different photoperiods and temperatures. A first strand cDNA was prepared from 10 pituitaries from male sticklebacks, and cDNA fragments were amplified by PCR using degenerated primers based on highly conserved regions of known teleost FSH-beta and LH-beta cDNA sequences. To obtain full-length cDNAs of FSH-beta and LH-beta, RACE amplifications were performed. The cDNA of FSH-beta was 540 by long, encoding a protein of 122 amino acids and LH-beta cDNA was 568 by long, encoding a protein of 148 amino acids. Of gonadotropin (GTH) beta-subunits published so far, those most similar to stickleback GTHs are found among other percomorph fishes, with amino acid similarities of 46-55% for FSH-beta and 75-77% for LH-beta subunits. The cloned cDNAs were used as probes to analyze LH and FSH mRNA expression in pituitaries from sticklebacks exposed to different photoperiods and temperatures. Two hundred males were divided into four groups and kept under different photoperiods and temperatures; light:dark (LD) 16:8, 20degreesC; LD 16:8, 7degreesC; LD 8:16, 20degreesC, and LD 8:16, 7degreesC. Red breeding colors and a marked kidney hypertrophy, androgen-dependent male secondary sexual characters in the stickleback, appeared in the group kept under LD 16:8 at 20degreesC, but not in the other groups. The kidney epithelium height (KEH) was significantly lower in the LD 8:16, 20degreesC group than in all other groups, and this was also the only group with active spermatogenesis. The LD 8:16 20degreesC had significantly lower expression of both FSH-beta and LH-beta than all other groups. Thus, both GTHs displayed their lowest expression when spermatogenesis was active and when low KEH indicated that androgens levels were at their lowest. (C) 2003 Elsevier Inc. All rights reserved.