Functional Identification and Characterization of the Diuretic Hormone 31 (DH31) Signaling System in the Green Shore Crab, Carcinus maenas

Abstract

The functional characterization of crustacean neuropeptides and their cognate receptors has not kept pace with the recent advances in sequence determination and, therefore, our understanding of the physiological roles of neuropeptides in this important arthropod sub-phylum is rather limited. We identified a candidate receptor-ligand pairing for diuretic hormone 31 (DH31) in a neural transcriptome of the crab, Carcinus maenas. In insects, DH31 plays species -specific but central roles in many facets of physiology, including fluid secretion, myoactivity, and gut peristalsis but little is known concerning its functions in crustaceans. The C. maenas DH31 transcript codes for a 147 amino acid prepropeptide, and a single receptor transcript translates to a secretin-like (Class B1) G protein-coupled receptor (GPCR). We used an in vitro aequorin luminescence Ca2+ mobilization assay to demonstrate that this candidate DH31R is activated byCarcinus and insect DH31s in a dose-dependent manner (EC50 15–30 nM). Whole mount immunohistochemical and in situ hybridization localization revealed extensive DH31 expressing neurons throughout the central nervous system, most notably in the abdominal ganglion where large, unpaired cells give rise to medial nerves, which terminate in extensive DH31 immunopositive dendritic fields intimately associated with oesophageal musculature. This system constitutes a large and hitherto undescribed neurohemal area adjacent to key muscle groups associated with the gastric system. DH31 expressing neurons were also seen in the cardiac, commissural, oesophageal, and stomatogastric ganglia and intense labeling was seen in dendrites innervating fore- and hindgut musculature but not with limb muscles. These labeling patterns, together with measurement of DH31R mRNA in the heart and hindgut, prompted us test the effects of DH31 on semi-isolated heart preparations. Cardiac superfusion with peptide evoked increased heart rates (10–100 nM). The neuroanatomical distribution of DH31 and its receptor transcripts, particularly that associated with gastric and cardiac musculature, coupled with the cardio- acceleratory effects of the peptide implicate this peptide in key myoactive roles, likely related to rhythmic coordination.

Characterization of G-protein coupled receptors from the blackback land crab Gecarcinus lateralis Y organ transcriptome over the molt cycle

“…CRF is structurally related to mammalian corticotropin, and is called Drome-DH31 in D. melanogaster [136]. DH31 was recently identified in several tissues of the green shore crab C. maenas and its function in rhythmic coordination was established [137]. CT peptide is structurally related to mammalian calcitonin-like peptide and is called Drome-DH44 in D. melanogaster [138].…”

Characterization of an evolutionarily conserved calcitonin signaling system in a lophotrochozoan, the Pacific oyster (Crassostrea gigas)

“…The arrangement of the oyster CTPs with two mature peptides is particularly intriguing as deuterostome CT-type peptide precursors contain either a single peptide (Muff et al, 1995;Sekiguchi et al, 2009) or two peptides, as observed in echinoderm CT-type precursors (Rowe et al, 2014;Suwansa-ard et al, 2018;Zandawala et al, 2017). In the protostomes, the annelid P. dumerilii CT and DH31 precursors (Conzelmann et al, 2013) and the DH31 and CT-A precursors in arthropods (Alexander et al, 2018;Christie, 2008;Christie et al, 2010;Coast et al, 2001;Gard et al, 2009;Li et al, 2007;Veenstra, 2014 comprise only one peptide. In molluscs, this tandem peptide arrangement is observed in bivalves (Zhang et al, 2018) and also in gastropods (Fig.…”

Signaling Pathways That Regulate the Crustacean Molting Gland

“…GPCRs are characterized by seven transmembrane domains, an external N-terminal domain, and a C-terminal cytosolic domain (25). These are divided among three large classes: rhodopsin-like (Class A), which represents the largest number of GPCRs; secretinlike (Class B); and metabotropic glutamate (Class C) (25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44).…”