Crustacean hyperglycemic hormones of two cold water crab species, Chionoecetes opilio and C. japonicus: Isolation of cDNA sequences and localization of CHH neuropeptide in eyestalk ganglia

• CHH cDNA sequences are isolated from cold water crab C. opilio and C. japonicus.
• The putative preproCHH sequence is identical in both species.
• ChoCHH expression is higher in the embryos at developing-eye stage than yolk stage and in the ES of males than females.
• The presence of Cho- and ChjCHH is found in the X-organ, axonal tract, and sinus gland.
• Cho- and ChjCHH cDNAs harbor putative translational regulatory sites: uORF and IRES.
• Translation of Cho- and ChjCHH may be tightly regulated.

Crustacean hyperglycemic hormone (CHH) is primarily known for its prototypical function in hyperglycemia which is induced by the release of CHH. The CHH release takes place as an adaptive response to the energy demands of the animals experiencing stressful environmental, physiological or behavioral conditions. Although >63 decapod CHH nucleotide sequences are known (GenBank), the majority of them is garnered from the species inhabiting shallow and warm water. In order to understand the adaptive role of CHH in Chionoecetes opilio and Chionoecetes japonicus inhabiting deep water environments, we first aimed for the isolation of the full-length cDNA sequence of CHH from the eyestalk ganglia of C. opilio (ChoCHH) and C. japonicus (ChjCHH) using degenerate PCR and 5′ and 3′ RACE. Cho- and ChjCHH cDNA sequences are identical in 5′ UTR and ORF with 100% sequence identity of the putative 138 aa of preproCHHs. The length of 3′ UTR ChjCHH cDNA sequence is 39 nucleotides shorter than that of ChoCHH. This is the first report in decapod crustaceans that two different species have the identical sequence of CHH. ChoCHH expression increases during embryogenesis of C. opilio and is significantly higher in adult males and females. C. japonicus males have slightly higher ChjCHH expression than C. opilio males, but no statistical difference. In both species, the immunostaining intensity of CHH is stronger in the sinus gland than that of X-organ cells. Future studies will enable us to gain better understanding of the comparative metabolic physiology and endocrinology of cold, deep water species of Chionoecetes spp.