Glycine facilitates transmitter release at developing synapses: a patch clamp study from Purkinje neurons of the newborn rat

Abstract

Synaptic currents in immature Purkinje cells from rats on postnatal days 0–14 ( P0–P14) were studied using whole-cell patch-electrodes applied to cerebellar slices (200 μm in thickness). Purkinje cells (held at −40 mV) showed excitatory postsynaptic currents (EPSCs) and inhibitory postsynaptic currents (IPSCs) spontaneously. From P2 to P12 the frequencies of miniature EPSCs and miniature IPSCs in the Purkinje cells increased by 10-fold or more, suggesting progressive formation of functional synapses during this period. Application of glycine (100 μM) to an immature Purkinje cell at P3–10 immediately increased the frequencies of both EPSCs and IPSCs. The effects of glycine showed maximum at P5–6 for EPSCs and at P9–10 for IPSCs and decreased thereafter. Facilitatory effects of glycine were suppressed by strychnine (1 μM), a specific blocker of the ionotropic glycine receptor, while the effects were also induced by other glycinergic agonists, including α- l-alanine (1 mM), l-serine (1 mM) and taurine (500 μM). The site of glycinergic effects was studied by removing the action potential generation in cerebellar slices. Following the addition of tetrodotoxin (TTX, 1 μM), the glycine-induced facilitation of EPSC almost disappeared, while that of IPSC remained (i.e. miniature IPSCs) and reached more than half of the value without TTX. These findings suggest that the ionotropic glycinergic receptors are expressed transiently but profoundly in the developing cerebellum, and that the distributions of these receptors causing excitation are different at excitatory and inhibitory presynaptic neurons. The glycine receptors may play distinct roles in the maturation and organization of cerebellar neural circuits.