Paraiso-Luna_2020_Development_147_

The endocannabinoid (eCB) system, via the cannabinoid CB(1) receptor, regulates neurodevelopment by controlling neural progenitor proliferation and neurogenesis. CB(1) receptor signalling in vivo drives corticofugal deep layer projection neuron development through the regulation of BCL11B and SATB2 transcription factors. Here, we investigated the role of eCB signalling in mouse pluripotent embryonic stem cell-derived neuronal differentiation. Characterization of the eCB system revealed increased expression of eCB-metabolizing enzymes, eCB ligands and CB(1) receptors during neuronal differentiation. CB(1) receptor knockdown inhibited neuronal differentiation of deep layer neurons and increased upper layer neuron generation, and this phenotype was rescued by CB(1) re-expression. Pharmacological regulation with CB(1) receptor agonists or elevation of eCB tone with a monoacylglycerol lipase inhibitor promoted neuronal differentiation of deep layer neurons at the expense of upper layer neurons. Patch-clamp analyses revealed that enhancing cannabinoid signalling facilitated neuronal differentiation and functionality. Noteworthy, incubation with CB(1) receptor agonists during human iPSC-derived cerebral organoid formation also promoted the expansion of BCL11B(+) neurons. These findings unveil a cell-autonomous role of eCB signalling that, via the CB(1) receptor, promotes mouse and human deep layer cortical neuron development.