Activation of type 1 cannabinoid receptor (CB1R) promotes neurogenesis in murine subventricular zone cell cultures

• Autor(a) principal: Xapelli, Sara Alves
• Data de Publicação: 2013
• Outros Autores: Agasse, Fabienne, Sardà-Arroyo, Laura, Bernardino, Liliana, Santos, Tiago, Ribeiro, Filipa F., Valero, Jorge, Bragança, José, Schitine, Clarissa, de Melo Reis, Ricardo A, Sebastião, Ana M., Malva, João O.
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10316/109774; https://doi.org/10.1371/journal.pone.0063529
• topic: Animals; Arachidonic Acids; Basic Helix-Loop-Helix Transcription Factors; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cerebral Ventricles; GABAergic Neurons; Gene Expression Regulation; Immunohistochemistry; Mice; Mice, Inbred C57BL; Multipotent Stem Cells; Nerve Tissue Proteins; Neurites; Neurons; Receptor, Cannabinoid, CB1; Stem Cells; Neurogenesis

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.