Two type I crustacean hyperglycemic hormone (CHH) genes in Morotoge shrimp (Pandalopsis japonica): Cloning and expression of eyestalk and pericardial organ isoforms produced by alternative splicing and a novel type I CHH with predicted structure shared with type II CHH peptides

Apr 25, 2012Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology

Two related hormones in Morotoge shrimp: gene versions from eye stalk and heart organs made by alternative splicing, including a new hormone type with features shared by a different hormone group

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Abstract

Three cDNAs encoding crustacean hyperglycemic hormone family members were isolated, with Pj-CHH1ES expressed more in the eyestalk X-organ/sinus gland complex.

Pj-CHH1ES and Pj-CHH1PO are derived from the same gene via alternative mRNA splicing, while Pj-CHH2 originates from a different gene.
The Pj-CHH1 and Pj-CHH2 genes contain four exons and three introns, indicating they may have arisen from gene duplication.
Pj-CHH1ES shows higher expression in the eyestalk X-organ/sinus gland complex compared to the gill.
Pj-CHH1PO has elevated expression levels in the eyestalk X-organ/sinus gland complex, brain, and abdominal ganglion.
Eyestalk ablation led to reduced expression of Pj-CHH1ES in the brain and Pj-CHH1PO and Pj-CHH2 in the thoracic ganglion.
Molecular modeling suggests Pj-CHH2 has a unique structure with a short alpha helix, indicating it may differ functionally from other type I CHHs.

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Crustacean hyperglycemic hormone (CHH) peptide family members play critical roles in growth and reproduction in decapods. Three cDNAs encoding CHH family members (Pj-CHH1ES, Pj-CHH1PO, and Pj-CHH2) were isolated by a combination of bioinformatic analysis and conventional cloning strategies. Pj-CHH1ES and Pj-CHH1PO were products of the same gene that were generated by alternative mRNA splicing, whereas Pj-CHH2 was the product of a second gene. The Pj-CHH1 and Pj-CHH2 genes had four exons and three introns, suggesting the two genes arose from gene duplication. The three cDNAs were classified in the type I CHH subfamily, as the deduced amino acid sequences had a CHH precursor-related peptide sequence positioned between the N-terminal signal sequence and C-terminal mature peptide sequence. The Pj-CHH1ES isoform was expressed at a higher level in the eyestalk X-organ/sinus gland (XO/SG) complex and at a lower level in the gill. The Pj-CHH1PO isoform was expressed at higher levels in the XO/SG complex, brain, abdominal ganglion, and thoracic ganglion and at a lower level in the epidermis. Pj-CHH2 was expressed at a higher level in the thoracic ganglion and at a lower level in the gill. Real-time polymerase chain reaction was used to quantify the effects of eyestalk ablation on the mRNA levels of the three Pj-CHHs in the brain, thoracic ganglion, and gill. Eyestalk ablation reduced expression of Pj-CHH1ES in the brain and Pj-CHH1PO and Pj-CHH2 in the thoracic ganglion. Sequence alignment of the Pj-CHHs with CHHs from other species indicated that Pj-CHH2 had an additional alanine at position #9 of the mature peptide. Molecular modeling showed that the Pj-CHH2 mature peptide had a short alpha helix (α1) in the N-terminal region, which is characteristic of type II CHHs. This suggests that Pj-CHH2 differs in function from other type I CHHs.

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Two type I crustacean hyperglycemic hormone (CHH) genes in Morotoge shrimp (Pandalopsis japonica): Cloning and expression of eyestalk and pericardial organ isoforms produced by alternative splicing and a novel type I CHH with predicted structure shared with type II CHH peptides

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