4.2 Article

Stimulation of molt by RNA interference of the molt-inhibiting hormone in the crayfish Cherax quadricarinatus

Journal

GENERAL AND COMPARATIVE ENDOCRINOLOGY
Volume 178, Issue 2, Pages 227-236

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ygcen.2012.05.007

Keywords

Cherax quadricarinatus; Molt-inhibiting hormone; RNAi; dsRNA; Molt cycle; Ecdysone

Funding

  1. UMBI-BARD Program [MB 8714 08]
  2. University of Maryland Biotechnology Institute
  3. United States-Israel Binational Agricultural Research and Development Fund
  4. Planning & Budget Commission of the Council for Higher Education (VATAT)

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In crustaceans, molting is known to be under the control of neuropeptide hormones synthesized and secreted from the eyestalk ganglia. While the role of molt-inhibiting hormone (MIH) in regulating molting has been described in several species using classical methods, an in vivo specific MIH targeted manipulation has not been described yet. In the present study, an MIH cDNA was isolated and sequenced from the eyestalk ganglia of the Australian freshwater red claw crayfish Cherax quadricarinatus (Cq) by 5' and 3' RACE. We analyzed the putative Cq-MIH based on sequence homology, a three dimensional structure model and transcript's tissue specificity. We further examined the involvement of Cq-MIH in the control of molt in the crayfish through RNAi by in vivo injections of Cq-MIH double-stranded RNA, which resulted in, similarly to eyestalk ablation, acceleration of molt cycles. This acceleration was reflected by a significant reduction (up to 32%) in molt interval and an increased rate in molt mineralization index (MMI), which correlated with the induction of ecdysteroid hormones compared to control. Altogether, this study provides a proof of function for the involvement of the Cq-MIH gene in molt regulation in the crayfish. (C) 2012 Elsevier Inc. All rights reserved.

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