Members of neuropeptide B/W signaling system have been predominantly detected and mapped within the CNS. In the rat, this system includes neuropeptide B (NPB), neuropeptide W (NPW) and their specific receptor NPBWR1. This signaling system has a wide spectrum of functions including a role in modulation of inflammatory pain and neuroendocrine functions. Expression of NPB, NPW and NPBWR1 in separate heart compartments, dorsal root ganglia (DRG) and stellate ganglia was proven by RT-qPCR, Western blot (WB) and immunofluorescence. Presence of mRNA for all tested genes was detected within all heart compartments and ganglia. The presence of proteins preproNPB, preproNPW and NPBWR1 was confirmed in all the chambers of heart by WB. Expression of preproNPW and preproNPB was proven in cardiac ganglionic cells obtained by laser capture microdissection. In immunofluorescence analysis, NPB immunoreactivity was detected in nerve fibers, some nerve cell bodies and smooth muscle within heart and both ganglia. NPW immunoreactivity was present in the nerve cell bodies and nerve fibers of heart ganglia. Weak nonhomogenous staining of cardiomyocytes was present within heart ventricles. NPBWR1 immunoreactivity was detected on cardiomyocytes and some nerve fibers. We confirmed the presence of NPB/W signaling system in heart, DRG and stellate ganglia by proteomic and genomic analyses.

Biomedical Center Faculty of Medicine in Pilsen Charles University 323 00 Pilsen Czech Republic

Department of Pharmacology and Toxicology Faculty of Medicine in Pilsen Charles University 323 00 Pilsen Czech Republic

Department of Physiology Faculty of Medicine in Pilsen Charles University 323 00 Pilsen Czech Republic

Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology Department of Chemistry Ugo Schiff University of Florence 500 19 Sesto Fiorentino Italy

Platform and CNRS BioCIS CY Cergy Paris Université 950 31 Cergy Pontoise CEDEX France

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Laser Capture Microdissection Coupled Capillary Immunoassay to Study the Expression of PCK-2 on Spatially-Resolved Islets of Rat Langerhans