Neuropeptide hormones from the eyestalks of Jasus Lalandii

Doctoral Thesis


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University of Cape Town

The X-organ sinus gland complex, situated in the eyestalks of decapod crustaceans, are known to be a source of a variety of neuropeptide hormones that regulate a number of diverse physiological processes. This neuroendocrine complex was investigated in 3 crustacean species, viz. the European shore crab Carcinus maellas, and 2 spiny lobster species Jasus lalandii and Panulirus homarus by means of tissue immunocytochemistry and an enzyme-linked immunosorbent assay (ELISA). Positive immunoreactions, associated with the X-organ - sinus gland system only, were obtained with antisera raised against crustacean hyperglycaemic hormone (cHH) of the American lobster (Homarus americanus), the Mexican crayfish (Procambarus bouvieri) and the edible crab (Cancer pagurus), as well as with antisera raised against vitellogenesis-inhibiting hormone (VIH) of the H. americanus and moult-inhibiting hormone (MIH) of C. pagurus. This is the first time that the immunolocalisation of these 3 hormones have been studied in a single crustacean species. The chief results of this comparative immunocytochemical study showed that (1) neuropeptide hormones of the shore crab and the 2 spiny lobster species were sufficiently homologous in primary structure to be recognised by the heterologous antisera, thus, an indication of conserved peptide structures across the species and infraorder boundaries; (2) preabsorbed complexes of purified peptides and antisera from the edible crab did not produce any immunoreactions in tissue immunocytochemistry, nor in ELISA, thus, indicating the specificity of the anti-cHH and anti-MIH sera; (3) the anti-VIH serum demonstrated the ability to bind epitopes on cHH and MIH peptides and is, thus, not a specific antiserum in this study; (4) there is co-localisation of cHH, MIH, VIH immunoreactivity in the eyestalk neuroendocrine complexes of all 3 species studied which suggests that the different peptide hormones can be synthesized in the same neuronal cell bodies. This co-localisation of neuropeptides in the eyestalk of J lalandii was confirmed by a double-staining immunoflourescence experiment, and finally (5) immunoreactivity of antisera raised against cHH of H. americanus and MIH of C. pagurus was associated with distinct and unique peak fractions, following reverse-phase high pressure liquid chromatographic (RP-HPLC) separation of sinus gland extracts from J. lalandii. A total of 6 neuropeptide hormones belonging to the cHH/MIH/VIH peptide family were isolated, functionally characterised and sequenced from extracts of sinus glands from the South African west coast rock lobster, Jasus lalandii. This is the first complete report on these peptides from any species belonging to the Palinuridae infraorder.

Bibliography: p. 242-264.