Insufficient developmental excitatory neuronal activity fails to foster establishment of normal levels of inhibitory neuronal activity

Abstract

The nervous system is composed of excitatory and inhibitory neurons. One major class of inhibitory neurons release the neurotransmitter γ-Aminobutyric acid (GABA). GABAergic inhibitory activity maintains the balance that is disrupted in conditions such as epilepsy. At least some GABAergic neurons are initially excitatory and undergo a developmental conversion to convert to inhibitory neurons. The mechanism(s) behind this conversion are thought to include a critical developmental increase in excitatory activity.To test this hypothesis, we subjected ex vivo developing neuronal networks on multi-electrode arrays to various stimulation and pharmacological regimens. Synaptic activity of networks initially consists of epileptiform-like high-amplitude individual “spikes”, which convert to organized bursts of activity over the course of approximately 1 month. Stimulation of networks with a digitized synaptic signal for 5days hastened the decrease of epileptiform activity. By contrast, stimulation for a single day delayed the appearance of bursts and instead increased epileptiform signaling.GABA treatment reduced total signals in unstimulated networks and networks stimulated for 5days, but instead increased signaling in networks stimulated for 1day. This increase was prevented by co-treatment with (2R)-amino-5-phosphonopentanoate and 6-cyano-7-nitroquinoxaline-2,3-dione, confirming that GABA invoked excitatory activity in networks stimulated for 1day. Glutamate increased signals in networks subjected to all stimulation regimens; the GABA receptor antagonist bicuculline prevented this increase only in networks stimulated for 1day. These latter findings are consistent with the induction of so-called “mixed” synapses (which release a combination of excitatory and inhibitory neurotransmitters) in networks stimulated for 1day, and support the hypothesis that a critical level of excitatory activity fosters the developmental transition of GABAergic neurons from excitatory to inhibitory.