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首页> 外文期刊>American Zoologist >Molecular cloning, expression, and tissue distribution of crustacean molt-inhibiting hormone
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Molecular cloning, expression, and tissue distribution of crustacean molt-inhibiting hormone

机译：甲壳类动物蜕皮抑制激素的分子克隆，表达及组织分布

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摘要

In crustaceans, secretion of ecdysteroid molting hormones by Y-organs is regulated by molt-inhibiting hormone (MIH), a neuropeptide produced by the X-organ/sinus gland complex of the eyestalks. The current review considers recent research on MIH, with a primary focus on MIH of brachyurans (crabs). New data on the production of recombinant MIH (rMIH) are also included. Available data indicate the MIH gene of brachyurans encodes a 113 amino acid prohormone composed of a 35 residue signal peptide and a 78 residue mature MIH. The primary structure of MIH is highly conserved among brachyurans. The MIH transcript is detectable in eyestalk neural ganglia throughout the molt cycle of the blue crab, Callinectes sapidus. Stage-specific changes in the abundance of MIH mRNA in C sapidus eyestalks are generally consistent with the hypothesis that MIH negatively regulates ecdysteroid production during the molt cycle. MIH transcripts have also been detected in the brain of two species. Recombinant MIH was produced using prokaryotic (pET vector/Escherichia coli) and eukaryotic (baculovirus/insect cells) expression systems. Recombinant MIH produced in E. coli was of the predicted size and was MIH immunoreactive; it did not have MIH bioactivity. Polyclonal antisera raised against the prokaryotically expressed rMIH bound specifically to neurosecretory cells in the X-organ, their associated axons, and axon terminals in the sinus gland. Recombinant MIH expressed using the baculovirus system was of the predicted size, was MIH immunoreactive, and inhibited ecdysteroid production by Y-organs in vitro.
机译：在甲壳类动物中，Y器官分泌蜕皮甾类蜕皮激素的过程受蜕皮抑制激素（MIH）的控制，MIH是由视线的X器官/窦腺复合体产生的一种神经肽。当前的审查考虑了最近对MIH的研究，主要关注短臂动物（蟹）的MIH。还包括有关重组MIH（rMIH）生产的新数据。现有数据表明，腕果动物的MIH基因编码113个氨基酸的激素，由35个残基的信号肽和78个残基的成熟MIH组成。 MIH的一级结构在短臂动物中高度保守。在蓝蟹Callinectes sapidus的整个蜕皮周期中，在眼柄神经节中可检测到MIH转录物。斜C眼柄中MIH mRNA丰度的阶段特定变化通常与以下假设相符：MIH在蜕皮周期中负面调节蜕皮类固醇的产生。在两个物种的大脑中也检测到了MIH转录本。使用原核（pET载体/大肠杆菌）和真核（杆状病毒/昆虫细胞）表达系统产生重组MIH。在大肠杆菌中产生的重组MIH具有预期大小，并且具有MIH免疫反应性。它没有MIH生物活性。抗原核表达的rMIH的多克隆抗血清特异性结合至X器官中的神经分泌细胞，其相关的轴突和窦腺的轴突末端。使用杆状病毒系统表达的重组MIH具有预期大小，具有MIH免疫反应性，并在体外抑制了Y-器官产生的蜕皮类固醇。

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