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Studies on crustacean hyperglycaemic hormone of the norway lobster nephrops norvegicus (L.)
journal contributionposted on 1994-01-01, 00:00 authored by Richard Smullen, M G Bentley
The Norway lobster, Nephrops norvegicus, displays a significant elevation of haemo-lymph glucose following the injection of sinus gland crude extracted, although the time course of this elevation appears to be different from other decapods studied. HPLC separation has been used to isolate and purify several neuropeptides which have hyperglycaemic properties from the sinus glands of Nephrops norvegicus. Enzyme linked immunosorbent assay (ELISA) experiments have shown that these peptides react with polyclonal rabbit antiserum raised against the CHH of the crayfish Orconectes limosus, while sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of these peptides has provided an estimation of their molecular weight. Pairs of oligonucleotide primers based on the sequence of the lobster Homarus americanus, complementary to regions of the CHH sequence, were used in the polymerase chain reactions (PCR). Complementary first strand DNA (cDNA) was synthesised from total Nephrops eyestalk RNA. PCR amplification produced a 100 bp and a 230 bp double stranded DNA product which was resolved by electrophoresis on a tris-borate-EDTA gel. The product size was compared with known standards and is being cloned into a vector in order to obtain a sequence so that a precise comparison to known CHH sequences can then be made. © 1994 Balaban.