Meeting Abstract
P3.77 Tuesday, Jan. 6 SEQUENCE ANALYSIS OF CRUSTACEAN HYPERGLYCEMIC HORMONE FROM TANNER CRAB CHIONOECETES BAIRDI TAMONE, SL*; CHUNG, JS; University of Alaska Southeast; UMBI-Center of Marine Biotechnology sltamone@uas.alaska.edu
Crustacean hyperglycemic hormone (CHH) is an eyestalk neuropeptide secreted from sinus glands that functions in the regulation of circulating glucose. This neuropetide is important for crustacean metabolism and should serve an important role in animals under environmental stress; animals that have increased metabolic rates to compensate for environmental change. Crustacean hyperglycemic peptide has been purified from multiple brachyuran crab species to date, but not from the cold water Tanner crab, Chionoecetes bairdi, a species found in Alaskan waters. The purpose of purifying and sequencing C. bairdi CHH is to establish tools for further study of cold water crab metabolic physiology. Cold water crabs such as those in the genus Chionoecetes are currently a good model for understanding the role that climate change and associated water temperature changes might have on metabolic physiology. Sinus glands were dissected from male C. bairdi and the neuropeptides were purified using reverse-phase HPLC. Two neuropeptides were identified as CHH with an enzyme-linked immunosorbant assay (ELISA) using a heterologous antibody specific for Callinectes sappidus CHH. The bioactivity of these two peaks was assessed by injecting CHH containing samples into eyestalk-ablated Tanner crabs and measuring subsequent circulating glucose. The full length cDNA sequence of the larger eyestalk CHH from C. bairdi was determined through amplification, cloning and sequencing of first strand cDNA and was subsequently constructed from overlapping clones. A phylogenetic analysis of amino acid sequence data from six brachyuran crab species showed C. bairdi CHH most closely related to the majid crab, Libinia emarginata (P55688). Future studies will include comparative metabolic physiology with the warm water crab C. sappidus.
