Structure and localization of mRNA encoding a pigment dispersing hormone (PDH) in the eyestalk of the crayfish Orconectes limosus

Abstract

The pigment-dispersing hormone (PDH) is produced in the eyestalks of Crustacea where it induces light-adapting movements of pigment in the compound eye and regulates the pigment dispersion in the chromatophores. To study this hormone at the mRNA level, we cloned and sequenced cDNA encoding PDH in the crayfish Orconectes Zimosus. The structure of the PDH preprohormone consists of a signal peptide, a PDH precursor-related peptide (PPRP) and the highly conserved PDH peptide at the carboxy-terminal end. In situ hybridization in combination with immunocytochemistry revealed four cell clusters expressing PDH in the optic ganglia of the eyestalk. Three clusters stained both with the PDH cRNA probe and the PDH antiserum, however, the perikarya in the lamina ganglionaris (LG) only stained with the PDH antiserum, suggesting the presence of a PDH-like peptide in the LG.

Identification of Neuropeptides from the Sinus Gland of the Crayfish Orconectes limosus Using Nanoscale On-line Liquid Chromatography Tandem Mass Spectrometry

“…Even using this relatively large amount of material, the PPRP, located between the signal peptide and the native PDH hormone (14), was not described to be present as a component of the SG. The question of whether the PPRP is transported and stored in the SG system has not been clarified yet (14).…”

“…Full characterization of these four CPRP variants remains still to be established. Under the standard conditions used for the on-line LC QTOF- (14), whereas in LC fractions 10 and 11 two new genetic variants of this hormone, PDHB and PDHC (Swiss-Prot accession nos. P83586 and P83587, respectively), were detected.…”

“…These neurohormones are classically considered as major physiological regulators involved in the control of carbohydrate metabolism and in various functions in development such as moult and reproduction (3, 6, 7). Additionally, the chromatophore-regulating hormones, like red pigmentconcentrating hormone (RPCH) and pigment-dispersing hormone (PDH), have been found in the XO-SG system of different crustacean species (8 -11).Different approaches, such as conventional chromatography, molecular cloning, and immunochemical methods, were used previously to investigate sinus gland components (12)(13)(14)(15)(16)(17). The limitations of these established methods to identify and characterize new peptides are the relatively low sensitivity in the case of conventional chromatography, the necessity of sufficient sequence homology in the case of molecular cloning, and the availability of specific antibodies in the case of immunochemical methods.…”

“…Different approaches, such as conventional chromatography, molecular cloning, and immunochemical methods, were used previously to investigate sinus gland components (12)(13)(14)(15)(16)(17). The limitations of these established methods to identify and characterize new peptides are the relatively low sensitivity in the case of conventional chromatography, the necessity of sufficient sequence homology in the case of molecular cloning, and the availability of specific antibodies in the case of immunochemical methods.…”

Cloning and differential expression of two β‐pigment‐dispersing hormone (β‐PDH) isoforms in the crab Cancer productus: Evidence for authentic β‐PDH as a local neurotransmitter and β‐PDH II as a humoral factor

“…The tissues were then incubated for approximately 72 hours in a 1:1,000 dilution of a rabbit polyclonal antibody generated against synthetic ␤-PDH (NSELINSILGLPKVMNDAamide) conjugated to bovine thyroglobulin via glutaraldehyde (Dr. K. Rao, University of West Florida, Pensacola, FL; Dircksen et al, 1987). This antibody has been used to map the distribution of ␤-PDH-like immunoreactivity in the nervous system of many crustaceans Mangerich and Keller, 1988;Mortin and Marder, 1991;de Kleijn et al, 1993), including both Cancer antennarius and Cancer borealis (Mortin and The Journal of Comparative Neurology Marder, 1991;Christie et al, 1995). Incubation of the antibody with synthetic ␤-PDH (10 -4 M) completely abolished all immunolabeling in both Cancer species (Mortin and Marder, 1991 Immunoresearch;.…”