{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:19:25Z","timestamp":1785403165495,"version":"3.56.0"},"posted":{"date-parts":[[2025,8,5]]},"group-title":"Cell Biology","reference-count":72,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,8,5]],"date-time":"2025-08-05T00:00:00Z","timestamp":1754352000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["DC019918"],"award-info":[{"award-number":["DC019918"]}],"id":[{"id":"https:\/\/ror.org\/04mhx6838","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,8,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Cochlear injury activates the resident macrophages (RM) and recruits the blood-circulating monocytes and monocyte-derived macrophages (Mo\/Mo-M), but their specific functions in the injured cochlea are unknown. It is well established that the chemokine fractalkine receptor (CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1), expressed by cochlear macrophages, influences the density of those macrophages and promotes synaptic repair and spiral ganglion neuron survival in the injured cochlea. As CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1 is expressed on both RM and Mo\/Mo-M, it remains unclear if CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1-expressing RM and Mo\/Mo-M are distinct and differentially promote SGN survival after cochlear injury. Here, we demonstrate the use of fate mapping via a tamoxifen-inducible CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1 mouse model (CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1\n                  <jats:sup>YFP\u2212CreERT2\/wildtype<\/jats:sup>\n                  :R26RFP) wherein CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1-expressing RM and Mo\/Mo-M are endogenously labeled with different fluorescent reporters to define the heterogeneity in cochlear macrophages regarding their origin, turnover, spatiotemporal distribution, morphology, and fate following a loud acoustic trauma. After 60 days of tamoxifen injections at 4 weeks of age, long-lived cochlear RM were YFP+ RFP+ with 98.0 \u00b1 1.7% recombinant efficiency, and short-lived blood-circulating CX\n                  <jats:sub>3<\/jats:sub>\n                  CR1 lineage (Mo\/Mo-M) were YFP+ RFP-with 2.5 \u00b1 1.1% recombinant efficiency. Following an acoustic trauma of 112 dB SPL at 8-16 kHz octave band for 2 hours, morphologically similar RM and Mo\/Mo-M were observed in the spiral ganglion, lamina, ligament, and around the sensory epithelium. Quantification of RM and Mo\/Mo-M in the spiral lamina and ganglion revealed distinct spatial and temporal distribution patterns. Furthermore, recruited Mo\/Mo-M expressed classical monocyte markers such as Ly6C and CCR2. Both RM and Mo\/Mo-M were positive for proliferation marker, Ki67, and negative for apoptotic marker, cleaved caspase-3, suggesting that the overall increase in macrophage numbers in the noise-injured cochlea is a contribution of both the proliferation of RM and recruitment of Mo\/Mo-M. Probing for blood-clotting protein, fibrinogen, showed its presence in the cochlea after acoustic trauma, suggesting vascular damage that positively and strongly correlated with the time course of recruitment of blood-circulating Mo\/Mo-M in the noise-injured cochlea. These data imply that macrophages in the noise-injured cochlea are heterogeneous regarding their ontogeny, distribution, and fate. They offer a robust tool to study the precise roles of resident and recruited macrophages in healthy and pathological ears.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Summary<\/jats:title>\n                  <jats:p>\n                    Using the novel CX\n                    <jats:sub>3<\/jats:sub>\n                    CR1 fate-mapping model, our data uncover diversity of macrophages with respect to their ontogeny, turnover, spatiotemporal distribution, and fate in the normal and noise-injured cochlea.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.08.04.668473","type":"posted-content","created":{"date-parts":[[2025,8,5]],"date-time":"2025-08-05T12:25:22Z","timestamp":1754396722000},"source":"Crossref","is-referenced-by-count":1,"title":["CX\n                  <sub>3<\/sub>\n                  CR1 Fate-Mapping In Vivo Distinguishes Cochlear Resident and Recruited Macrophages After Acoustic Trauma"],"prefix":"10.64898","author":[{"given":"Sree Varshini","family":"Murali","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew R","family":"Stothert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elyssa","family":"Pereyra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lyudmila V","family":"Batalkina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tejbeer","family":"Kaur","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025081611300851000_2025.08.04.668473v2.1","doi-asserted-by":"publisher","DOI":"10.1038\/nn2014"},{"key":"2025081611300851000_2025.08.04.668473v2.2","doi-asserted-by":"crossref","first-page":"516","DOI":"10.3390\/brainsci13030516","article-title":"Identification and Characterization of TMEM119-Positive Cells in the Postnatal and Adult Murine Cochlea","volume":"13","year":"2023","journal-title":"Brain Sciences"},{"key":"2025081611300851000_2025.08.04.668473v2.3","doi-asserted-by":"publisher","DOI":"10.1002\/glia.22315"},{"key":"2025081611300851000_2025.08.04.668473v2.4","doi-asserted-by":"crossref","unstructured":"Chiot, A. , Felgner, M. 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