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The Journal of Experimental Biology 206, 979-988 (2003)
doi: 10.1242/jeb.00178

Putative involvement of crustacean hyperglycemic hormone isoforms in the neuroendocrine mediation of osmoregulation in the crayfish Astacus leptodactylus

Laetitia Serrano¹, Gaëlle Blanvillain¹, Daniel Soyez², Guy Charmantier¹, Evelyse Grousset¹, Fabien Aujoulat¹ and Céline Spanings-Pierrot^1,*

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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