Cultured cerebellar neurons: endogenous and exogenous components of Purkinje cell activity and membrane response to putative transmitters.

Modified explant cultures of fetal rat cerebellum were developed for electrophysiological and pharmacological studies, at the membrane level, of Purkinje neurons. The goals of the present series of experiments were to identify possible endogenous and exogenous components to the electrical activity of Purkinje neurons, to assess the sensitivity of these neurons to putative excitatory and inhibitory neurotransmitters, and to characterize the membrane response to the transmitters. Intracellular recordings were made from Purkinje neurons, identified on a morphological basis, using conventional electrophysiological techniques. Virtually all Purkinje neurons displayed spontaneous activity. A contribution of both endogenous and exogenous components to the spontaneous activity was indicated by alterations in the pattern and amount of activity when the membrane potential was varied and by the characteristics of the individual potentials themselves. Several types of activity were considered to be endogenous: the most common type consisted of pacemaker-like potentials which generated a pattern of firing similar to that characterized as simple spike activity in previous in vivo studies; another type of endogenous activity consisted of large membrane depolarizations that evoked one or two spikes. These depolarizing responses were similar to the membrane response generated by climbing fiber input to Purkinje cells in vivo. The exogenous components to the spontaneous activity consisted of synaptic potentials including excitatory (EPSPs) and inhibitory (IPSPs) synaptic potentials and biphasic EPSP/IPSPs. Several putative transmitters thought to mediate these synaptic potentials were tested by focal micropressure application to determine if they could mimic the action of the endogenous transmitters. The putative transmitter glutamate depolarized the cultured Purkinje neurons and evoked action potentials, characteristics which were displayed by the excitatory synaptic potentials. The putative inhibitory transmitter GABA hyperpolarized the cultured Purkinje neurons and depressed activity, characteristics which were displayed by the inhibitory synaptic potentials. The putative inhibitory transmitters glycine and taurine were ineffective. Norepinephrine, the transmitter mediating the inhibitory input from the locus coeruleus to Purkinje neurons, was also tested. When applied in the microM range, NE effects were variable. When applied in the mM range, NE depressed the spontaneous activity in a manner suggestive of a presynaptic action.