Molecular characterization of gonad‐inhibiting hormone of Penaeus monodon and elucidation of its inhibitory role in vitellogenin expression by RNA interference

Jan 29, 2008The FEBS journal

Molecular study of gonad-inhibiting hormone in Penaeus monodon and its role in reducing egg yolk protein production using gene silencing

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Abstract

Pem-GIH cDNA is 861 bp in size and encodes a peptide that may inhibit vitellogenin synthesis in shrimp.

The cDNA for Pem-GIH includes a single open reading frame (ORF) of 288 bp.
Pem-GIH consists of a 17-residue signal peptide and a mature peptide region of 79 amino acids.
Transcript levels of Pem-GIH were found in multiple tissues, including the eyestalk and brain.
RNA interference targeting Pem-GIH led to decreased transcript levels in both eyestalk and abdominal nerve cord cultures.
A notable increase in vitellogenin transcript levels was observed in the ovaries of GIH-knockdown shrimp, suggesting that Pem-GIH negatively regulates vitellogenin expression.

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One of the important peptide hormones that control reproduction in crustaceans is gonad-inhibiting hormone (GIH). GIH is known to modulate gonad maturation by inhibiting synthesis of vitellogenin (Vg), the precursor of yolk proteins. In this study, a cDNA encoding a GIH (Pem-GIH) from the eyestalk of Penaeus monodon was cloned using RT-PCR and RACE techniques. Pem-GIH cDNA is 861 bp in size with a single ORF of 288 bp. The deduced Pem-GIH consists of a 17-residue signal peptide and a mature peptide region of 79 amino acids with features typical of type II peptide hormones from the CHH family. Pem-GIH transcript was detected in eyestalk, brain, thoracic and abdominal nerve cords of adult P. monodon. The gonad-inhibiting activity of Pem-GIH was investigated using the RNA interference technique. Double-stranded RNA, corresponding to the mature Pem-GIH sequence, can trigger a decrease in Pem-GIH transcript levels both in eyestalk ganglia and abdominal nerve cord explant culture and in female P. monodon broodstock. The conspicuous increase in Vg transcript level in the ovary of GIH-knockdown shrimp suggests a negative influence for Pem-GIH on Vg gene expression, and thus implies its role as a gonad-inhibiting hormone. This is the first report to demonstrate the use of double-stranded RNA to elucidate the function of GIH in P. monodon.

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Molecular characterization of gonad‐inhibiting hormone of Penaeus monodon and elucidation of its inhibitory role in vitellogenin expression by RNA interference

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