Frontiers in Neuroscience (Jan 2012)
A role for the cannabinoid 1 receptor in neuronal differentiation of adult spinal cord progenitors in vitro is revealed through pharmacological inhibition and genetic deletion
Abstract
In contrast to the adult brain, the adult spinal cord is a non-neurogenic environment. Understanding how to manipulate the spinal cord environment to promote the formation of new neurons is an attractive therapeutic strategy for spinal cord injury and disease. The cannabinoid 1 receptor (CB1R) has been implicated as a modulator of neural progenitor cell proliferation and fate specification in the brain; however, no evidence exists for modulation of adult spinal cord progenitor cells. Using adult rat spinal cord primary cultures, we demonstrated that CB1R antagonism with AM251 significantly decreased the number of Nestin(+) cells, and increased the number of βIII tubulin(+) and DCX(+) cells, indicative of neuronal differentiation. AM251’s effect was blocked by co-application of the CB1R agonists, WIN 55,212-2 or ACEA. Consistent with our hypothesis, the chronic absence of CB1R via the use of knock-out (CB1-/-) mice resulted in significantly higher levels of DCX(+) cells compared to wild type (CB1+/+) cultures, indicative of enhanced neuronal differentiation in CB1-/- spinal cords. Moreover, AM251 promoted neuronal differentiation in CB1+/+, but not in CB1-/- cultures. Since CB1R modulates synaptic transmission, and synaptic transmission has been shown to influence progenitor cell fate, we evaluated whether AM251-induced neuronal differentiation was affected by chronic inactivity. Interestingly, either the presence of the voltage-dependent sodium channel blocker tetrodotoxin (TTX), or the removal of mature neurons, inhibited the AM251-induced increase in DCX (+) cells. In summary, antagonism or absence of CB1R promotes neuronal differentiation in adult spinal cord cultures, and this action appears to require TTX-sensitive neuronal activity.
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