Glycine and GABA, the dominant inhibitory neurotransmitters in the CNS, assume a depolarizing role in early development, leading to increased cytoplasmic Ca 2ϩ levels and action potentials. The effect is thought to be of some significance for maturation. The depolarization is caused by Cl Ϫ efflux, and chloride transporters contribute to the phenomenon by raising the intracellular Cl

Summary Ribbon synapses convey sustained and phasic excitatory drive within retinal microcircuits. However, the properties of retinal inhibitory synapses are less well known. AII-amacrine cells are interneurons in the retina that exhibit large glycinergic synapses at their dendritic lobular appendages. Using membrane capacitance measurements we observe robust exocytosis elicited by the opening of L-type Ca2+ channels located on the lobular appendages. Two pools of synaptic vesicles were detected: a small, rapidly releasable pool and a larger and more slowly releasable pool. Depending on the stimulus, either paired-pulse depression or facilitation could be elicited. During early postnatal maturation, the coupling of the exocytosis Ca2+-sensor to Ca2+ channel becomes tighter. Light-evoked depolarizations of the AII-amacrine cell elicited exocytosis that was graded to light intensity. Our results suggest that AII-amacrine cell synapses are capable of providing both phasic and sustained inhibitory input to their postsynaptic partners without the benefit of synaptic ribbons.

Most fast-acting neurotransmitters are rapidly cleared from synaptic regions. This feature isolates synaptic sites, rendering the timecourse of synaptic responses independent of the number of active synapses. We describe a striking exception at glycinergic synapses on granule cells of the rat dorsal cochlear nucleus. The duration of IPSCs was dependent on the number of presynaptic axons that were stimulated and on the number of vesicles released from each axon. Increasing stimulus number or frequency, or blocking glycine uptake, slowed synaptic decays, while a low-affinity competitive antagonist of GlyRs accelerated IPSC decay. These effects could be explained by unique features of GlyRs when activated by pooling of glycine across synapses. Functionally, increasing the number of IPSPs markedly lengthened the period of spike inhibition following cessation of presynaptic stimulation. Thus, temporal properties of inhibition can be controlled by activity levels in multiple presynaptic cells or by adjusting release probability at individual synapses.

Thyroid hormone (TH) deficiency during perinatal life causes a multitude of functional and morphological deficits in the brain. In rats and mice, TH dependency of neural maturation is particularly evident during the first 1-2 weeks of postnatal development. During the same period, synaptic transmission via the inhibitory transmitters glycine and GABA changes from excitatory depolarizing effects to inhibitory hyperpolarizing ones in most neurons [depolarizing-hyperpolarizing (D/H) shift]. The D/H shift is caused by the activation of the K(+)-Cl(-) co-transporter KCC2 which extrudes Cl(-) from the cytosol, thus generating an inward-directed electrochemical Cl(-) gradient. Here we analyzed whether the D/H shift and, consequently, the onset of inhibitory neurotransmission are influenced by TH. Gramicidin perforated-patch recordings from auditory brainstem neurons of experimentally hypothyroid rats revealed depolarizing glycine effects until postnatal day (P)11, i.e. almost 1 week longer than in control rats, in which the D/H shift occurred at approximately P5-6. Likewise, until P12-13 the equilibrium potential E(Gly) in hypothyroids was more positive than the membrane resting potential. Normal E(Gly) could be restored upon TH substitution in P11-12 hypothyroids. These data demonstrate a disturbed Cl(-) homeostasis following TH deficiency and point to a delayed onset of synaptic inhibition. Interestingly, immunohistochemistry demonstrated an unchanged KCC2 distribution in hypothyroids, implying that TH deficiency did not affect KCC2 gene expression but may have impaired the functional status of KCC2. Hippocampal neurons of hypothyroid P16-17 rats also demonstrated an impaired Cl(-) homeostasis, indicating that TH may have promoted the D/H shift and maturation of synaptic inhibition throughout the brain.