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LSO neurons weigh interaural level differences (ILDs) through precise integration of glutamatergic excitation from the cochlear nucleus (CN) and glycinergic inhibition from the medial nucleus of the trapezoid body (MNTB). Sound sources can be localized even during sustained perception, an accomplishment that requires robust neurotransmission. Virtually nothing is known about the sustained performance and the temporal precision of MNTB\u2013LSO inputs after postnatal day (P)12 (time of hearing onset) and whether acoustic experience guides development. Here we performed whole\u2010cell patch\u2010clamp recordings to investigate neurotransmission of single MNTB\u2010LSO fibres upon sustained electrical stimulation (1\u2013200\u00a0Hz\/60\u00a0s) at P11 and P38 in wild\u2010type (WT) and deaf otoferlin (<jats:italic>Otof<\/jats:italic>) knock\u2010out (KO) mice. At P11, WT and KO inputs performed remarkably similarly. In WTs, the performance increased drastically between P11 and P38, e.g. manifested by an 8 to 11\u2010fold higher replenishment rate (RR) of synaptic vesicles and action potential robustness. Together, these changes resulted in reliable and highly precise neurotransmission at frequencies \u2264100\u00a0Hz. In contrast, KO inputs performed similarly at both ages, implying impaired synaptic maturation. Computational modelling confirmed the empirical observations and established a reduced RR per release site for P38 KOs. In conclusion, acoustic experience appears to contribute massively to the development of reliable neurotransmission, thereby forming the basis for effective ILD detection. Collectively, our results provide novel insights into experience\u2010dependent maturation of inhibitory neurotransmission and auditory circuits at the synaptic level.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key points<\/jats:title><jats:p><jats:list list-type=\"bullet\">\n<jats:list-item><jats:p>Inhibitory glycinergic inputs from the medial nucleus of the trapezoid body (MNTB) to the lateral superior olive (LSO) are involved in sound localization. This brainstem circuit performs reliably throughout life. How such reliability develops is unknown.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>Here we investigated the role of acoustic experience on the functional maturation of MNTB\u2010LSO inputs at juvenile (postnatal day P11) and young adult ages (P38) employing deaf mice lacking otoferlin (KO).<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>We analysed neurotransmission at single MNTB\u2010LSO fibres in acute brainstem slices employing prolonged high\u2010frequency stimulation (1\u2013200\u00a0Hz\/60\u00a0s).<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>At P11, KO inputs still performed normally, as manifested by normal synaptic attenuation, fidelity, replenishment rate, temporal precision and action potential robustness. Between P11 and P38, several synaptic parameters increased substantially in wild\u2010type mice, collectively resulting in high\u2010fidelity and temporally precise neurotransmission. In contrast, maturation of synaptic fidelity was largely absent in KOs after P11.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>Collectively, reliable neurotransmission at inhibitory MNTB\u2013LSO inputs develops under the guidance of acoustic experience.<\/jats:p><\/jats:list-item>\n<\/jats:list><\/jats:p><\/jats:sec>","DOI":"10.1113\/jp280403","type":"journal-article","created":{"date-parts":[[2022,4,19]],"date-time":"2022-04-19T19:34:13Z","timestamp":1650396853000},"page":"2461-2497","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Development of synaptic fidelity and action potential robustness at an inhibitory sound localization circuit: effects of otoferlin\u2010related deafness"],"prefix":"10.1113","volume":"600","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4389-6268","authenticated-orcid":false,"given":"Nicolas I.C.","family":"M\u00fcller","sequence":"first","affiliation":[{"name":"Animal 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