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These signals have been shown to regulate not only the refinement of neural circuits along the auditory pathway towards functional maturity, but also the maturation of the hair cells into sensory receptors. Although the origin and interplay of these Ca\n                    <jats:sup>2+<\/jats:sup>\n                    signals during cochlear development have recently been investigated in live mice, the impact of anaesthesia on in vivo functional measurements was not explored. Here, we investigate the effects of different anaesthetic regimes (ketamine and xylazine; 2.5% isoflurane; and 1.0%\u20131.5% isoflurane with the sedative acepromazine) that provided an effective unconsciousness to perform the surgery and Ca\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010imaging recordings from the intact cochlea of live mice. The IHCs, supporting cells and spiral ganglion neuron terminals onto the IHCs showed spontaneous Ca\n                    <jats:sup>2+<\/jats:sup>\n                    \u2010dependent activity under all anaesthetic regimes, with a few significant differences observed between conditions. Calcium waves from supporting cells synchronized the activity of IHCs. Moreover, we found that the endocochlear potential, which is crucial for cochlear function, was unaffected by the different anaesthetics. However, low concentrations of isoflurane produced the most stable recordings of vital physiological signs in mice, including heart rate and breathing rate. Although all anaesthetic regimes tested appeared to be suitable for performing Ca\n                    <jats:sup>2+<\/jats:sup>\n                    imaging from the cochlea of pre\u2010hearing live mice, a low concentration of isoflurane (1.0%\u20131.5%), combined with the pre\u2010anaesthetic sedative acepromazine and oxygenation, represents the most suitable approach to maintain a stable and long\u2010lasting depth of anaesthesia.\n                  <\/jats:p>","DOI":"10.1113\/ep093267","type":"journal-article","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T17:06:49Z","timestamp":1760116009000},"page":"1253-1267","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Spontaneous Ca\n                    <sup>2+<\/sup>\n                    signals in the developing mammalian cochlea of live mice under different anaesthetic regimes"],"prefix":"10.1113","volume":"111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7691-5635","authenticated-orcid":false,"given":"Francesca","family":"De Faveri","sequence":"first","affiliation":[{"name":"School of Biosciences University of Sheffield  Sheffield UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8770-7628","authenticated-orcid":false,"given":"Walter","family":"Marcotti","sequence":"additional","affiliation":[{"name":"School of Biosciences University of Sheffield  Sheffield UK"},{"name":"Neuroscience Institute University of Sheffield  Sheffield UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5366-341X","authenticated-orcid":false,"given":"Federico","family":"Ceriani","sequence":"additional","affiliation":[{"name":"School of Biosciences University of Sheffield  Sheffield UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,10,10]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0800793105"},{"key":"e_1_2_9_3_1","first-page":"443","article-title":"Optimization of intraperitoneal injection anesthesia in mice: Drugs, dosages, adverse effects, and anesthesia depth","volume":"51","author":"Arras M.","year":"2001","journal-title":"Comparative Medicine"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.2178-20.2020"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1097\/ACO.0000000000000916"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1205"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.21-13-04593.2001"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1017114108"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0304-3940(92)90162-Z"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.15252\/embj.2022112118"},{"key":"e_1_2_9_11_1","first-page":"71","article-title":"Differential actions of isoflurane and ketamine\u2010based anaesthetics on cochlear function in the mouse","volume":"292","author":"Cederholm J. 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