Endocannabinoid (eCB) signaling serves as an on-demand neuroprotective system. eCBs are produced postsynaptically in response to depolarization or activation of metabotropic glutamate receptors (mGluRs) and act on presynaptic cannabinoid receptor-1 to suppress synaptic transmission. Here, we examined the effects of epileptiform activity on these two forms of eCB signaling in hippocampal cultures. Treatment with bicuculline and 4-aminopyridine (Bic + 4-AP), which induced burst firing, inhibited metabotropic-induced suppression of excitation (MSE) and prolonged the duration of depolarization-induced suppression of excitation (DSE). The Homer family of proteins provides a scaffold for signaling molecules including mGluRs. It is known that seizures induce the expression of the short Homer isoform 1a (H1a) that acts in a dominant negative manner to uncouple Homer scaffolds. Bic + 4-AP treatment increased H1a mRNA. A group I mGluR antagonist blocked the Bic + 4-AP-evoked increase in burst firing, the increase in H1a expression, and the inhibition of MSE. Bic + 4-AP treatment reduced mGluR-mediated Ca2+ mobilization from inositol trisphosphate-sensitive stores relative to untreated cells. Expression of H1a, but not a mutant form that cannot bind Homer ligands, mimicked Bic + 4-AP inhibition of MSE and mGluR-mediated Ca2+ mobilization. In cells expressing shRNA targeted to Homer 1 mRNA, Bic + 4-AP did not affect mGluR-mediated Ca2+ release. Furthermore, knockdown of H1a prevented the inhibition of MSE induced by Bic + 4-AP. Thus, an epileptic stimulus increased H1a expression, which subsequently uncoupled mGluR-mediated eCB production. These results indicate that seizure activity modulates eCB-mediated synaptic plasticity, suggesting a changing role for the eCB system following exposure to aberrant patterns of excitatory synaptic activity. Highlights: Epileptiform activity inhibits mGluR-mediated endocannabinoid production. Epileptiform activity prolongs depolarization-induced endocannabinoid production. An epileptiform stimulus increased Homer 1a expression to uncouple mGluR signaling. Seizures alter the mechanism of endocannabinoid-mediated synaptic plasticity.
Bibliographical noteFunding Information:
This study was supported by National Institutes of Health grants DA07304 and DA11806 and National Science Foundation grant IOS0814549 to S.A. Thayer.
- Homer proteins
- Metabotropic glutamate receptor