title: Localization and production of peptide endocannabinoids in the rodent CNS and adrenal medulla.
abstract: The endocannabinoid system (ECS) comprises the cannabinoid receptors CB1 and CB2 and their endogenous arachidonic acid-derived agonists 2-arachidonoyl glycerol and anandamide, which play important neuromodulatory roles. Recently, a novel class of negative allosteric CB1 receptor peptide ligands, hemopressin-like peptides derived from alpha hemoglobin, has been described, with yet unknown origin and function in the CNS. Using monoclonal antibodies we now identified the localization of RVD-hemopressin (pepcan-12) and N-terminally extended peptide endocannabinoids (pepcans) in the CNS and determined their neuronal origin. Immunohistochemical analyses in rodents revealed distinctive and specific staining in major groups of noradrenergic neurons, including the locus coeruleus (LC), A1, A5 and A7 neurons, which appear to be major sites of production/release in the CNS. No staining was detected in dopaminergic neurons. Peptidergic axons were seen throughout the brain (notably hippocampus and cerebral cortex) and spinal cord, indicative of anterograde axonal transport of pepcans. Intriguingly, the chromaffin cells in the adrenal medulla were also strongly stained for pepcans. We found specific co-expression of pepcans with galanin, both in the LC and adrenal gland. Using LC-MS/MS, pepcan-12 was only detected in non-perfused brain (∼ 40 pmol/g), suggesting that in the CNS it is secreted and present in extracellular compartments. In adrenal glands, significantly more pepcan-12 (400-700 pmol/g) was measured in both non-perfused and perfused tissues. Thus, chromaffin cells may be a major production site of pepcan-12 found in blood. These data uncover important areas of peptide endocannabinoid occurrence with exclusive noradrenergic immunohistochemical staining, opening new doors to investigate their potential physiological function in the ECS. This article is part of the Special Issue entitled 'Fluorescent Tools in Neuropharmacology'.
authors.affiliation: Institute of Biochemistry and Molecular Medicine, University of Bern, Bühlstrasse 28, CH-3012 Bern, Switzerland.
authors.forename: Stefanie C
authors.initials: SC
authors.lastname: Hofer
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
authors.forename: William T
authors.initials: WT
authors.lastname: Ralvenius
authors.affiliation: Institute of Biochemistry and Molecular Medicine, University of Bern, Bühlstrasse 28, CH-3012 Bern, Switzerland.
authors.forename: M Salomé
authors.initials: MS
authors.lastname: Gachet
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Fritschy
authors.affiliation: Institute of Pharmacology and Toxicology, University of Zürich, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland; Institute of Pharmaceutical Chemistry, Swiss Federal Institute of Technology (ETH) Zürich, Vladimir-Prelog-Weg 4, 8093 Zürich, CH-8057 Zürich, Switzerland.
authors.forename: Hanns Ulrich
authors.initials: HU
authors.lastname: Zeilhofer
authors.affiliation: Institute of Biochemistry and Molecular Medicine, University of Bern, Bühlstrasse 28, CH-3012 Bern, Switzerland. Electronic address: gertsch@ibmm.unibe.ch.
authors.forename: Jürg
authors.initials: J
authors.lastname: Gertsch
mesh_terms: D000313:Adrenal Medulla / Q000254:growth & development / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001596:Benzylamines / Q000494:pharmacology; D002490:Central Nervous System / Q000254:growth & development / Q000378:metabolism*; D002853:Chromatography, Liquid; D003078:Colchicine / Q000494:pharmacology; D063388:Endocannabinoids / Q000737:chemistry* / Q000378:metabolism*; D005260:Female; D006454:Hemoglobins; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D014179:Neurotransmitter Uptake Inhibitors / Q000494:pharmacology; D010446:Peptide Fragments; D010455:Peptides / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D053719:Tandem Mass Spectrometry; D050257:Tubulin Modulators / Q000494:pharmacology
chemical_list: D001596:Benzylamines; D063388:Endocannabinoids; D006454:Hemoglobins; D014179:Neurotransmitter Uptake Inhibitors; D010446:Peptide Fragments; D010455:Peptides; C000590773:RVD-hemopressin, human; D050257:Tubulin Modulators; C012102:DSP 4; D003078:Colchicine
keywords: Adrenal gland; CB1 receptor; Hemopressin; Locus coeruleus; Neuropeptides; Noradrenergic system; Pepcan-12; Pepcans