A novel CHH gene from the Pacific white shrimp Litopenaeus vannamei was characterized and found highly expressed in gut and less in eyestalk and other extra-eyestalk tissues

Feb 11, 2016Gene

New CHH Gene in Pacific White Shrimp Is Highly Active in the Gut and Less in the Eyestalk and Other Body Tissues

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Abstract

A new gene containing 4 exons and 3 introns was isolated from Litopenaeus vannamei.

Two full-length transcripts of the crustacean hyperglycemic hormone (CHH) were identified, measuring 1578bp in males and 974bp in females.
The length variation of transcripts is attributed to two polyadenylation sites in the 3'UTR that produce different termination signals.
A unique protein encoded by the gene can be classified as a type I CHH subfamily member, despite some structural differences.
Phylogenetic analysis places the peptide closer to type II CHHs rather than within type I.
The gene is expressed in neuroendocrine organs associated with CHH production, as well as in gill, gut, pericardial cavity, and male reproductive organs.

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The crustacean hyperglycemic hormone (CHH) family is an important group of neuropeptides involved in controlling growth, reproduction, and stress response in decapod species. In this study, a new gene containing 4 exons-3 introns flanked by canonical 5'-GT-AG-3' intron splice-site junctions was isolated from Litopenaeus vannamei. Two full length transcripts of this CHH were isolated from eyestalk and pericardial tissue of males and females using rapid amplification of cDNA ends (RACE). Transcripts sequences were 1578bp in length in males pericardial tissues and in males and females eyestalk with 100% identity, but the transcript isolated from females pericardial tissues was shorter (974bp). The differences in transcripts length is a result of two polyadenylation sites present in the 3'UTR resulting in two transcription termination signals. Transcript sequences encoded one unique protein that can be classified as type I CHH subfamily because of the 4 exons and 3 introns structure, although the CPRP region is not-well conserved and there is no amidation in the C-terminal of the deduced amino acid sequence. Furthermore, there is a glycine inserted in the mature peptide not at position 12 as in type II CHHs but after amino acid 31 and the phylogenetic analysis did not group the peptide within type I, but closer to type II CHHs. We demonstrated by endpoint-PCR, qPCR, and in situ hybridization (ISH), that this gene is expressed in neuroendocrine organs known to express CHHs in penaeid shrimp, including X-organ and optic nerve in eyestalk, supraesophageal ganglion (SoG), but it is also expressed in other organs as gill, gut, pericardial cavity, as well as in terminal ampoule or spermatophore and vas deferens of males.

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A novel CHH gene from the Pacific white shrimp Litopenaeus vannamei was characterized and found highly expressed in gut and less in eyestalk and other extra-eyestalk tissues

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