Endocannabinoid signalling in stem cells and cerebral organoids drives differentiation to deep layer projection neurons via CB1 receptors

Development. 2020 Dec 23;147(24):dev192161. doi: 10.1242/dev.192161.


The endocannabinoid (eCB) system, via the cannabinoid CB1 receptor, regulates neurodevelopment by controlling neural progenitor proliferation and neurogenesis. CB1 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 CB1 receptors during neuronal differentiation. CB1 receptor knockdown inhibited neuronal differentiation of deep layer neurons and increased upper layer neuron generation, and this phenotype was rescued by CB1 re-expression. Pharmacological regulation with CB1 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 CB1 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 CB1 receptor, promotes mouse and human deep layer cortical neuron development.

Keywords: 2-Arachidonoylglycerol; Cerebral organoid; Cortical development; Endocannabinoids; Neurogenesis; Projection neuron; Tetrahydrocannabinol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics*
  • Cell Proliferation / drug effects
  • Cerebellum / growth & development
  • Embryonic Development / genetics
  • Endocannabinoids / agonists
  • Endocannabinoids / genetics
  • Endocannabinoids / metabolism
  • Gene Expression Regulation, Developmental / genetics
  • Humans
  • Induced Pluripotent Stem Cells / drug effects
  • Matrix Attachment Region Binding Proteins / genetics*
  • Mice
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism
  • Neurogenesis / drug effects
  • Neurons / metabolism*
  • Organoids / growth & development
  • Receptor, Cannabinoid, CB1 / genetics*
  • Repressor Proteins / genetics*
  • Signal Transduction / genetics
  • Transcription Factors / genetics*
  • Tumor Suppressor Proteins / genetics*


  • Bcl11b protein, mouse
  • Endocannabinoids
  • Matrix Attachment Region Binding Proteins
  • Receptor, Cannabinoid, CB1
  • Repressor Proteins
  • SATB2 protein, mouse
  • Transcription Factors
  • Tumor Suppressor Proteins

Associated data

  • Dryad/10.5061/dryad.h9w0vt4g9