title: Activation of type 1 cannabinoid receptor (CB1R) promotes neurogenesis in murine subventricular zone cell cultures.
abstract: The endocannabinoid system has been implicated in the modulation of adult neurogenesis. Here, we describe the effect of type 1 cannabinoid receptor (CB1R) activation on self-renewal, proliferation and neuronal differentiation in mouse neonatal subventricular zone (SVZ) stem/progenitor cell cultures. Expression of CB1R was detected in SVZ-derived immature cells (Nestin-positive), neurons and astrocytes. Stimulation of the CB1R by (R)-(+)-Methanandamide (R-m-AEA) increased self-renewal of SVZ cells, as assessed by counting the number of secondary neurospheres and the number of Sox2+/+ cell pairs, an effect blocked by Notch pathway inhibition. Moreover, R-m-AEA treatment for 48 h, increased proliferation as assessed by BrdU incorporation assay, an effect mediated by activation of MAPK-ERK and AKT pathways. Surprisingly, stimulation of CB1R by R-m-AEA also promoted neuronal differentiation (without affecting glial differentiation), at 7 days, as shown by counting the number of NeuN-positive neurons in the cultures. Moreover, by monitoring intracellular calcium concentrations ([Ca(2+)]i) in single cells following KCl and histamine stimuli, a method that allows the functional evaluation of neuronal differentiation, we observed an increase in neuronal-like cells. This proneurogenic effect was blocked when SVZ cells were co-incubated with R-m-AEA and the CB1R antagonist AM 251, for 7 days, thus indicating that this effect involves CB1R activation. In accordance with an effect on neuronal differentiation and maturation, R-m-AEA also increased neurite growth, as evaluated by quantifying and measuring the number of MAP2-positive processes. Taken together, these results demonstrate that CB1R activation induces proliferation, self-renewal and neuronal differentiation from mouse neonatal SVZ cell cultures.
authors.affiliation: Center for Neuroscience and Cell Biology of Coimbra, University of Coimbra, Coimbra, Portugal.
authors.forename: Sara
authors.initials: S
authors.lastname: Xapelli
authors.forename: Fabienne
authors.initials: F
authors.lastname: Agasse
authors.forename: Laura
authors.initials: L
authors.lastname: Sardà-Arroyo
authors.forename: Liliana
authors.initials: L
authors.lastname: Bernardino
authors.forename: Tiago
authors.initials: T
authors.lastname: Santos
authors.forename: Filipa F
authors.initials: FF
authors.lastname: Ribeiro
authors.forename: Jorge
authors.initials: J
authors.lastname: Valero
authors.forename: José
authors.initials: J
authors.lastname: Bragança
authors.forename: Clarissa
authors.initials: C
authors.lastname: Schitine
authors.forename: Ricardo A
authors.initials: RA
authors.lastname: de Melo Reis
authors.forename: Ana M
authors.initials: AM
authors.lastname: Sebastião
authors.forename: João O
authors.initials: JO
authors.lastname: Malva
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002552:Cerebral Ventricles / Q000166:cytology*; D059330:GABAergic Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000378:metabolism; D055495:Neurogenesis / Q000187:drug effects / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D043884:Receptor, Cannabinoid, CB1 / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; D043884:Receptor, Cannabinoid, CB1; C088155:methanandamide; C486336:Neurog1 protein, mouse