Novel Protocol for the Chemical Synthesis of Crustacean Hyperglycemic Hormone Analogues — An Efficient Experimental Tool for Studying Their Functions

Mosco, Alessandro; Zlatev, Vientsislav; Guarnaccia, Corrado; Pongor, Sándor; Campanella, Antonella; Zahariev, Sotir; Giulianini, Piero Giulio

doi:10.1371/journal.pone.0030052

Cited by 9 publications

(19 citation statements)

References 47 publications

(64 reference statements)

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“…Serrano et al (Serrano et al, 2003) demonstrated that, in P. leptodactylus, a more prominent increase of osmolarity and sodium concentration in the circulation was caused by D-CHH than by L-CHH. Finally, a different time course and intensity of increase in circulating glucose was triggered by the two isomers in P. leptodactylus (Lebaupain et al, 2012;Mosco et al, 2012;Serrano et al, 2003). Earlier hyperglycemia was induced by L-CHH but it lasted for a shorter period and was less intensive than that induced by D-CHH.…”

Section: Introductionmentioning

confidence: 91%

“…A broader time scale of 48 h was examined in experiment 2 and results are presented in supplementary material Fig. S1, revealing the typical time course and intensity caused by each of the applied isomers (Mosco et al, 2012).…”

Section: Morphology and Physiology Of The Experimental Femalesmentioning

confidence: 99%

“…No external abdominally incubated ova were observed in females participating in either of the experiments. The source recombinant CHHs and their synthesis procedure were described by Mosco et al (Mosco et al, 2012). The technical procedure of injection into the circulation, the evaluation of circulating glucose level, the female seasonal reproductive status, and the carapace length and molt stage were detailed previously .…”

Section: Crayfish Maintenance and Experimental Proceduresmentioning

confidence: 99%

“…Each of the two isomers is synthesized in specific Xorgan cells (Soyez et al, 1998) and the L-isomer of H. americanus (Soyez et al, 1997) and Orconectes limosus (Ollivaux and Soyez, 2000) is about three times more abundant than the D-isomer. Recently, the two CHH enantiomers of Pontastacus leptodactylus were chemically synthesized and the physiological effects of the recombinant proteins have been demonstrated (Lebaupain et al, 2012;Mosco et al, 2012). Access to large amounts of synthesized isomers obviates the tedious extraction and purification procedures of the native isomers from the XOSG.…”

Section: Introductionmentioning

confidence: 99%

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Different transcription regulation routes are exerted by L- and D-amino acid enantiomers of peptide hormones

Tom

Manfrin

Mosco

et al. 2014

Journal of Experimental Biology

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Conversion of one or more amino acids in eukaryotic peptides to the D-enantiomer configuration is catalyzed by specific L/D-peptide isomerases and it is a poorly investigated post-translational modification. No common modified amino acid or specific modified position has been recognized, and mechanisms underlying changes in the peptide function provided by this conversion are not widely studied. The 72 amino acid crustacean hyperglycemic hormone (CHH) in Astacidea crustaceans exhibits a co-existence of two peptide enantiomers with either D-or L-phenylalanine as their third residue. It is a pleiotropic hormone regulating several physiological processes in different target tissues and along different time scales. CHH enantiomers differently affect time courses and intensities of examined processes. The short-term effects of the two isomers on gene expression were examined in the hepatopancreas, gills, hemocytes and muscles of the astacid Pontastacus leptodactylus. Gene expression in muscles and hemocytes was not affected by either of the isomers. Two modes of action for CHH were elucidated in the hepatopancreas and the gills: specific gene induction in both organs by D-CHH, and targeted attenuation caused by both enantiomers in the gills. Consequently, a two-receptor system is proposed for conveying the effect of the two CHH isomers.

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Section: Introductionmentioning

confidence: 91%

Section: Morphology and Physiology Of The Experimental Femalesmentioning

confidence: 99%

Section: Crayfish Maintenance and Experimental Proceduresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Different transcription regulation routes are exerted by L- and D-amino acid enantiomers of peptide hormones

Tom

Manfrin

Mosco

et al. 2014

Journal of Experimental Biology

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“…Although P. leptodactylus is not considered a model organism, it is one of the most studied species among decapods with about 143 published papers regarding its physiology [1], [36], [37], [38], its resistance to different types of pollution and stress conditions [39], [40], [41] and concerning the cHH and its variety of functions [42], [43], [44], [45]. Unfortunately this variety of studies does not reflect the fast progress of genomics and transcriptomics.…”

Section: Introductionmentioning

confidence: 99%

Application of D-Crustacean Hyperglycemic Hormone Induces Peptidases Transcription and Suppresses Glycolysis-Related Transcripts in the Hepatopancreas of the Crayfish Pontastacus leptodactylus — Results of a Transcriptomic Study

et al. 2013

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The crustacean Hyperglycemic Hormone (cHH) is a neuropeptide present in many decapods. Two different chiral isomers are simultaneously present in Astacid crayfish and their specific biological functions are still poorly understood. The present study is aimed at better understanding the potentially different effect of each of the isomers on the hepatopancreatic gene expression profile in the crayfish Pontastacus leptodactylus, in the context of short term hyperglycemia. Hence, two different chemically synthesized cHH enantiomers, containing either L- or D-Phe3, were injected to the circulation of intermolt females following removal of their X organ-Sinus gland complex. The effects triggered by the injection of the two alternate isomers were detected after one hour through measurement of circulating glucose levels. Triggered changes of the transcriptome expression profile in the hepatopancreas were analyzed by RNA-seq. A whole transcriptome shotgun sequence assembly provided the assumedly complete transcriptome of P. leptodactylus hepatopancreas, followed by RNA-seq analysis of changes in the expression level of many genes caused by the application of each of the hormone isomers. Circulating glucose levels were much higher in response to the D-isoform than to the L-isoform injection, one hour from injection. Similarly, the RNA-seq analysis confirmed a stronger effect on gene expression following the administration of D-cHH, while just limited alterations were caused by the L-isomer. These findings demonstrated a more prominent short term effect of the D-cHH on the transcription profile and shed light on the effect of the D-isomer on specific functional gene groups. Another contribution of the study is the construction of a de novo assembly of the hepatopancreas transcriptome, consisting of 39,935 contigs, that dramatically increases the molecular information available for this species and for crustaceans in general, providing an efficient tool for studying gene expression patterns in this organ.

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Functional analysis of a mutated analogue of the crustacean hyperglycaemic hormone from the crayfish Pontastacus leptodactylus

et al. 2015

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The crustacean hyperglycaemic hormone (CHH), a pleiotropic neuropeptide, belongs to a family of structurally related peptides, having six cysteine residues in conserved positions forming three disulphide bridges, and regulating several physiological processes in crustaceans and insects. Structure-activity studies have shown that amidation of the C-terminus is important to confer biological activity to CHH. In this study we investigated the function of the d-Phe(3) of the N-terminal motif of Pontastacus leptodactylus CHH by a mutational analysis. The d-Phe in position 3 was substituted by a d-Ala and the functionality of the mutated analogue (Glp-d-A-CHH) was tested by in vivo biological assays. The mutated analogue resulted far less active than its wild-type counterparts, either in d- (Glp-d-CHH) or l- (Glp-l-CHH) configuration. These results suggest that Phe(3) is essential for the biological activity of P. leptodactylus CHH, demonstrating that also the N-terminus is involved in the binding with the receptor, and identifying in the Phe(3) a hot spot for the peptide-receptor binding.

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Novel Protocol for the Chemical Synthesis of Crustacean Hyperglycemic Hormone Analogues — An Efficient Experimental Tool for Studying Their Functions

Cited by 9 publications

References 47 publications

Different transcription regulation routes are exerted by L- and D-amino acid enantiomers of peptide hormones

Different transcription regulation routes are exerted by L- and D-amino acid enantiomers of peptide hormones

Application of D-Crustacean Hyperglycemic Hormone Induces Peptidases Transcription and Suppresses Glycolysis-Related Transcripts in the Hepatopancreas of the Crayfish Pontastacus leptodactylus — Results of a Transcriptomic Study

Functional analysis of a mutated analogue of the crustacean hyperglycaemic hormone from the crayfish Pontastacus leptodactylus

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