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In the present study, we applied a multiple\u2010site optical recording technique with a voltage\u2010sensitive dye and surveyed the developmental organization of the auditory system in the mouse embryo. Stimulation of the cochlear (auditory) nerve elicited excitatory postsynaptic potential (EPSP)\u2010related optical responses in the lateral brainstem, which corresponded to the auditory sensory nucleus, namely, the cochlear nucleus. The EPSP was mediated by glutamate and mainly dependent on\n                    <jats:italic>N<\/jats:italic>\n                    \u2010methyl\u2010d\u2010aspartate (NMDA) receptors. The EPSP was first detected at E (embryonic day) 12, indicating that functional connections between the periphery and neurons in the cochlear nucleus are established at this stage. At later developmental stages, cochlear nerve stimulation elicited additional postsynaptic responses in the ipsilateral and contralateral ventral brainstem, which corresponded to the higher center of the auditory pathway, the superior olivary complex (SOC). The EPSP in the SOC was detected from E15 in normal physiological solution and E14 in a Mg\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010free solution. Thus, the synaptic connections between the cochlear nucleus and the SOC are established by E14, but they are suppressed by the Mg\n                    <jats:sup>2+<\/jats:sup>\n                    block on the NMDA receptors before E15. Overall, our results suggest that postsynaptic responses in the cochlear nucleus and the SOC exhibit much earlier than previously reported, and functional synapses are generated soon after the arrival of afferent fibers and before morphological differentiation of the brainstem nuclei has been completed.\n                  <\/jats:p>","DOI":"10.1111\/ejn.70106","type":"journal-article","created":{"date-parts":[[2025,4,12]],"date-time":"2025-04-12T04:28:21Z","timestamp":1744432101000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Functional Development of the Auditory Brainstem Nuclei During Embryogenesis of the Mouse Revealed by Optical Recording With a Voltage\u2010Sensitive Dye"],"prefix":"10.1111","volume":"61","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8613-0978","authenticated-orcid":false,"given":"Yoko","family":"Momose\u2010Sato","sequence":"first","affiliation":[{"name":"Department of Nutrition and Dietetics College of Nutrition, Kanto Gakuin University  Yokohama Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9565-209X","authenticated-orcid":false,"given":"Katsushige","family":"Sato","sequence":"additional","affiliation":[{"name":"Department of Health and Nutrition Sciences, Faculty of Human Health Komazawa Women's University  Tokyo Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,4,12]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bb.13.060184.001203"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pneurobio.2011.01.004"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1980.sp013443"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2178-20.2020"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00239.2022"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0165-3806(87)90140-4"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0165-3806(90)90164-T"},{"key":"e_1_2_10_9_1","first-page":"417","volume-title":"Nomina Anatomica Avium","author":"Breazile J. 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