Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus

Abstract Molecular profiling of inner ear neurons has broadened the classification of the primary afferents that support neural coding for hearing and balance. To extend spatiotemporal characterization of auditory and vestibular neuron diversity, we established a transgenic reporter mouse model (Prp...

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Main Authors: Lily J. Pearson, Jeremy L. Pinyon, Jennie M. E. Cederholm, Georg von Jonquieres, Florence Bartlett, Xabier Vázquez-Campos, Fabien Delerue, Lars M. Ittner, Gary D. Housley
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-94011-3
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author Lily J. Pearson
Jeremy L. Pinyon
Jennie M. E. Cederholm
Georg von Jonquieres
Florence Bartlett
Xabier Vázquez-Campos
Fabien Delerue
Lars M. Ittner
Gary D. Housley
author_facet Lily J. Pearson
Jeremy L. Pinyon
Jennie M. E. Cederholm
Georg von Jonquieres
Florence Bartlett
Xabier Vázquez-Campos
Fabien Delerue
Lars M. Ittner
Gary D. Housley
author_sort Lily J. Pearson
collection DOAJ
description Abstract Molecular profiling of inner ear neurons has broadened the classification of the primary afferents that support neural coding for hearing and balance. To extend spatiotemporal characterization of auditory and vestibular neuron diversity, we established a transgenic reporter mouse model (Prph p-mCherry), where elements of the peripherin promoter (Prph p) drive expression of the mCherry fluorescent reporter. Type III intermediate filament protein peripherin expression is a marker for type II spiral ganglion neurons (SGN) that innervate the cochlear outer hair cells, and the small diameter ‘bouton’ vestibular ganglion neurons (VGN) innervating the type II vestibular hair cells. Using Nanopore genome sequencing, the integration site of the transgene construct was identified within the class III metabotropic glutamate receptor 8 gene (Grm8, chromosome 6). Use of CUBIC / PEGASOS clearing of early postnatal to adult inner ear tissues enabled in situ 3D spatial localization of a dispersed population of cochlear mCherry + ve SGN, with highest expression and density in the hook (high frequency encoding) basal region. Of these mCherry + ve SGN, type II SGN (peripherin-immunopositive) were all co-labeled in the basal region, but the majority of the overall mCherry-delineated SGN auditory subpopulation were type I SGN innervating inner hair cells. In the VGN, mCherry + ve neurons represented ~ 15% of the adult population, dispersed as a small diameter subpopulation throughout both the inferior and superior VGN regions. These findings resolve heterogeneous type I and type II cochlear SGN subpopulations, particularly in the structurally complex hook region, and further differentiate vestibular primary afferents across postnatal development.
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spelling doaj-art-3d84ca12b1ec47008715dc9b43cb570e2025-08-20T02:10:12ZengNature PortfolioScientific Reports2045-23222025-03-0115112010.1038/s41598-025-94011-3Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locusLily J. Pearson0Jeremy L. Pinyon1Jennie M. E. Cederholm2Georg von Jonquieres3Florence Bartlett4Xabier Vázquez-Campos5Fabien Delerue6Lars M. Ittner7Gary D. Housley8Translational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, UNSWTranslational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, UNSWTranslational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, UNSWTranslational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, UNSWKGLMF Mark Wainwright Analytical Centre, UNSWSchool of Biotechnology and Biomolecular Sciences, UNSWDepartment of Genetics, The University of Texas MD Anderson Cancer CenterFaculty of Medicine, Health and Human Sciences, Department of Biomedical Sciences, Dementia Research Centre, Macquarie UniversityTranslational Neuroscience Facility, Department of Physiology, School of Biomedical Sciences, UNSWAbstract Molecular profiling of inner ear neurons has broadened the classification of the primary afferents that support neural coding for hearing and balance. To extend spatiotemporal characterization of auditory and vestibular neuron diversity, we established a transgenic reporter mouse model (Prph p-mCherry), where elements of the peripherin promoter (Prph p) drive expression of the mCherry fluorescent reporter. Type III intermediate filament protein peripherin expression is a marker for type II spiral ganglion neurons (SGN) that innervate the cochlear outer hair cells, and the small diameter ‘bouton’ vestibular ganglion neurons (VGN) innervating the type II vestibular hair cells. Using Nanopore genome sequencing, the integration site of the transgene construct was identified within the class III metabotropic glutamate receptor 8 gene (Grm8, chromosome 6). Use of CUBIC / PEGASOS clearing of early postnatal to adult inner ear tissues enabled in situ 3D spatial localization of a dispersed population of cochlear mCherry + ve SGN, with highest expression and density in the hook (high frequency encoding) basal region. Of these mCherry + ve SGN, type II SGN (peripherin-immunopositive) were all co-labeled in the basal region, but the majority of the overall mCherry-delineated SGN auditory subpopulation were type I SGN innervating inner hair cells. In the VGN, mCherry + ve neurons represented ~ 15% of the adult population, dispersed as a small diameter subpopulation throughout both the inferior and superior VGN regions. These findings resolve heterogeneous type I and type II cochlear SGN subpopulations, particularly in the structurally complex hook region, and further differentiate vestibular primary afferents across postnatal development.https://doi.org/10.1038/s41598-025-94011-3
spellingShingle Lily J. Pearson
Jeremy L. Pinyon
Jennie M. E. Cederholm
Georg von Jonquieres
Florence Bartlett
Xabier Vázquez-Campos
Fabien Delerue
Lars M. Ittner
Gary D. Housley
Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
Scientific Reports
title Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
title_full Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
title_fullStr Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
title_full_unstemmed Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
title_short Developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin-promoter reporter within the Grm8 locus
title_sort developmental differentiation of mouse inner ear neuron subpopulations resolved with a peripherin promoter reporter within the grm8 locus
url https://doi.org/10.1038/s41598-025-94011-3
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