کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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5901650 | 1156129 | 2011 | 8 صفحه PDF | دانلود رایگان |
Molt-inhibiting hormone (MIH), a member of the crustacean hyperglycemic hormone (CHH) family, inhibits the synthesis of ecdysteroid in Y-organ (YO) and plays a significant role in the regulation of molting and growth of crustaceans. A complete cDNA sequence encoding MIH (Ers-MIH, GenBank Accession No.: DQ341280) was cloned from eyestalk of Chinese mitten crab (Eriocheir sinensis) by 5â² and 3â² RACEs and PCR cloning. The full-length cDNA consists of 1457Â bp with a 330Â bp open reading frame, encoding 110 amino acids, containing a 75 amino acid mature peptide. The deduced amino acid sequence contains a typical CHH domain. Transcripts of Ers-MIH mRNA were detected in eyestalk by Northern blotting. The production of purified recombinant Ers-MIH (rErs-MIH) expressed in Escherichia coli was 0.3Â g/L. The LC-ESI-MS analysis showed that two peptide fragments of the recombinant protein were identical to the deduced amino acid sequence of Ers-MIH. By in vitro assay on E. sinensis YOs, a cGMP mediated suppression of rErs-MIH on ecdysteroidogenesis could be observed. Accumulation of cGMP in YOs showed a concentration-dependent manner within 0.01-1Â nmol/mL of rErs-MIH; ecdysteroid secretion was inhibited significantly at the range of 0.01-100Â nmol/mL rErs-MIH; furthermore, a significant inhibition effect on ecdysteroid releasing was shown when cGMP analog (8-Br-cGMP) concentration rose up to 100Â nmol/mL. This study would facilitate to investigate the roles of MIH in molt cycle regulation.
⺠A MIH (Ers-MIH) gene was cloned from eyestalk ganglia of Chinese mitten crab (Eriocheir sinensis). ⺠Transcripts of Ers-MIH mRNA were detected in eyestalk by Northern blotting. ⺠Recombinant Ers-MIH (rErs-MIH) was expressed in Escherichia coli. ⺠By in vitro assay on E. sinensis YOs, a cGMP mediated suppression of rErs-MIH on ecdysteroidogenesis was verified.
Journal: General and Comparative Endocrinology - Volume 173, Issue 3, 15 September 2011, Pages 467-474