Glycinergic neurotransmission is mediated via inhibitory glycine receptors (GlyRs) which are heterogeneous during development. Electrophysiological studies performed on recombinant GlyRs have identified different pharmacological properties and attributed them to differences in their subunit composition. Here, we report on age-related changes in the response properties of native GlyRs in the mammalian brain. Whole-cell patch-clamp recordings were obtained from neurons of the medial nucleus of the trapezoid body (MNTB), a major relay station in the mammalian auditory brainstem. Experiments were performed in acute medullary slices of rats between postnatal day (P) 1 and P15, a period during which synapse maturation occurs. Glycine-induced currents were present throughout the period under investigation and displayed age-related modifications in their amplitude, kinetic characteristics, and sensitivity to drugs. Current amplitudes and GlyR desensitization behavior increased with age. The alpha 1 subunit-specific GlyR antagonist cyanotriphenylborate (CTB) was barely effective in reducing glycine-induced currents during the first few postnatal days, yet a significant increase of the inhibitory effect occurred after the first postnatal week. This finding indicates that alpha 1 subunit-containing GlyRs become expressed only postnatally in the MNTB. Picrotoxin, which most effectively blocks recombinant alpha 2-homooligomers, reduced glycine-induced currents in neonatal MNTB neurons, suggesting that alpha 2-homooligomers may form native GlyR isoforms. Our results show that the physiology and pharmacology of GlyRs in the auditory brainstem underlie age-related changes which are most probably produced through a replacement of "neonatal" alpha 2 subunits with "adult" alpha 1 subunits.