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Putative involvement of crustacean hyperglycemic hormone isoforms in the neuroendocrine mediation of osmoregulation in the crayfish Astacus leptodactylus
1 Laboratoire Génome, Populations, Interactions, Adaptation, UMR
5000, Equipe Adaptation Ecophysiologique et Ontogenèse,
Université Montpellier II, Place E. Bataillon, CP 092, 34095
Montpellier Cédex 05, France
2 Groupe Biogenèse des Peptides Isomères, UMR CNRS Physiologie
et Physiopathologie, Université Paris VI, CC 256, 7 Quai Saint-Bernard,
75252 Paris Cédex 05, France
* Author for correspondence (e-mail: pierrot{at}univ-montp2.fr)
Accepted 19 December 2002
This study investigates the involvement of eyestalk neuroendocrine factors on osmoregulation in the crayfish Astacus leptodactylus maintained in freshwater. Eyestalk removal was followed by a significant decrease in hemolymph osmolality and Na+ concentration and by a 50% increase in mass after one molting cycle. Several neurohormones have been isolated from the sinus gland through high-performance liquid chromatography (HPLC), and different crustacean hyperglycemic hormone (CHH)-related peptides, including stereoisomers (L-CHH and D-Phe3 CHH), have been identified by direct enzyme-linked immunosorbent assay (ELISA). A glucose quantification bioassay demonstrated a strong hyperglycemic activity following injection of the immunoreactive chromatographic fractions and showed that the D-Phe3 CHH was the most efficient. Destalked crayfish were then injected with purified CHH HPLC fractions. The D-Phe3 CHH fraction significantly increased the hemolymph osmolality and Na+ content 24 h after injection. Two other CHH-related peptides caused a smaller increase in Na+ concentration. No significant variation was observed in hemolymph Cl- concentration following injection of any of the CHH isoforms. These results constitute the first observation of the effects of a CHH isoform, specifically the D-Phe3 CHH, on osmoregulatory parameters in a freshwater crustacean. The effects of eyestalk ablation and CHH injection on osmoregulation and the identification of different CHH-related peptides and isoforms in crustaceans are discussed.
Key words: Crustacea, crayfish, Astacus leptodactylus, eyestalk, crustacean hyperglycemic hormone, osmoregulation, neuropeptide, CHH, isoform, D-Phe3 CHH
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