Neuronal development in the inner ear is initiated by expression of the proneural basic Helix-Loop-Helix (bHLH) transcription factor Neurogenin1 that specifies neuronal precursors in the otocyst. The initial specification of the neuroblasts within the otic epithelium is followed by the expression of an additional bHLH factor, Neurod1. Although NEUROD1 is essential for inner ear neuronal development, the different aspects of the temporal and spatial requirements of NEUROD1 for the inner ear and, mainly, for auditory neuron development are not fully understood. In this study, using Foxg1Cre for the early elimination of Neurod1 in the mouse otocyst, we showed that Neurod1 deletion results in a massive reduction of differentiating neurons in the otic ganglion at E10.5, and in the diminished vestibular and rudimental spiral ganglia at E13.5. Attenuated neuronal development was associated with reduced and disorganized sensory epithelia, formation of ectopic hair cells, and the shortened cochlea in the inner ear. Central projections of inner ear neurons with conditional Neurod1 deletion are reduced, unsegregated, disorganized, and interconnecting the vestibular and auditory systems. In line with decreased afferent input from auditory neurons, the volume of cochlear nuclei was reduced by 60% in Neurod1 mutant mice. Finally, our data demonstrate that early elimination of Neurod1 affects the neuronal lineage potential and alters the generation of inner ear neurons and cochlear afferents with a profound effect on the first auditory nuclei, the cochlear nuclei.

Department of Biology University of Iowa Iowa City IA United States

Department of Physiology and Cell Biology Institute for Neuroscience University of Nevada Reno NV United States

Laboratory of Molecular Pathogenesis Institute of Biotechnology CAS Vestec Czechia

Zobrazit více v PubMed

Abrams S. R., Reiter J. F. (2021). Ciliary Hedgehog Signaling Regulates Cell Survival to Build the Facial Midline. Elife 10, e68558. 10.7554/elife.68558 PubMed DOI PMC

Bermingham N. A., Hassan B. A., Price S. D., Vollrath M. A., Ben-Arie N., Eatock R. A., et al. (1999). Math1 : An Essential Gene for the Generation of Inner Ear Hair Cells. Science 284, 1837–1841. 10.1126/science.284.5421.1837 PubMed DOI

Bérubé N. G., Mangelsdorf M., Jagla M., Vanderluit J., Garrick D., Gibbons R. J., et al. (2005). The Chromatin-Remodeling Protein ATRX Is Critical for Neuronal Survival During Corticogenesis. J. Clin. Invest. 115, 258–267. 10.1172/jci200522329 PubMed DOI PMC

Bohuslavova R., Smolik O., Malfatti J., Berkova Z., Novakova Z., Saudek F., et al. (2021). NEUROD1 Is Required for the Early Alpha and Beta Endocrine Differentiation in the Pancreas. Int. J. Mol. Sci. 22, 6713. 10.3390/ijms22136713 PubMed DOI PMC

Brulet R., Matsuda T., Zhang L., Miranda C., Giacca M., Kaspar B. K., et al. (2017). NEUROD1 Instructs Neuronal Conversion in Non-reactive Astrocytes. Stem Cel Rep. 8, 1506–1515. 10.1016/j.stemcr.2017.04.013 PubMed DOI PMC

Cherry T. J., Wang S., Bormuth I., Schwab M., Olson J., Cepko C. L. (2011). NeuroD Factors Regulate Cell Fate and Neurite Stratification in the Developing Retina. J. Neurosci. 31, 7365–7379. 10.1523/jneurosci.2555-10.2011 PubMed DOI PMC

Dastidar S. G., Landrieu P. M. Z., D'Mello S. R. (2011). FoxG1 Promotes the Survival of Postmitotic Neurons. J. Neurosci. 31, 402–413. 10.1523/jneurosci.2897-10.2011 PubMed DOI PMC

Duggan C. D., DeMaria S., Baudhuin A., Stafford D., Ngai J. (2008). Foxg1 Is Required for Development of the Vertebrate Olfactory System. J. Neurosci. 28, 5229–5239. 10.1523/jneurosci.1134-08.2008 PubMed DOI PMC

Duncan J. S., Fritzsch B. (2013). Continued Expression of GATA3 Is Necessary for Cochlear Neurosensory Development. PLoS One 8, e62046. 10.1371/journal.pone.0062046 PubMed DOI PMC

Dvorakova M., Macova I., Bohuslavova R., Anderova M., Fritzsch B., Pavlinkova G. (2020). Early Ear Neuronal Development, but Not Olfactory or Lens Development, Can Proceed Without SOX2. Dev. Biol. 457, 43–56. 10.1016/j.ydbio.2019.09.003 PubMed DOI PMC

Eagleson K. L., Schlueter McFadyen-Ketchum L. J., Ahrens E. T., Mills P. H., Does M. D., Nickols J., et al. (2007). Disruption of Foxg1 Expression by Knock-In of Cre Recombinase: Effects on the Development of the Mouse Telencephalon. Neuroscience 148, 385–399. 10.1016/j.neuroscience.2007.06.012 PubMed DOI PMC

Elliott K. L., Fritzsch B., Duncan J. S. (2018). Evolutionary and Developmental Biology Provide Insights into the Regeneration of Organ of Corti Hair Cells. Front Cel Neurosci 12, 252. 10.3389/fncel.2018.00252 PubMed DOI PMC

Elliott K. L., Pavlínková G., Chizhikov V. V., Yamoah E. N., Fritzsch B. (2021). Development in the Mammalian Auditory System Depends on Transcription Factors. Ijms 22, 4189. 10.3390/ijms22084189 PubMed DOI PMC

Evsen L., Sugahara S., Uchikawa M., Kondoh H., Wu D. K. (2013). Progression of Neurogenesis in the Inner Ear Requires Inhibition of Sox2 Transcription by Neurogenin1 and Neurod1. J. Neurosci. 33, 3879–3890. 10.1523/jneurosci.4030-12.2013 PubMed DOI PMC

Filova I., Dvorakova M., Bohuslavova R., Pavlinek A., Elliott K. L., Vochyanova S., et al. (2020). Combined Atoh1 and Neurod1 Deletion Reveals Autonomous Growth of Auditory Nerve Fibers. Mol. Neurobiol. 57, 5307–5323. 10.1007/s12035-020-02092-0 PubMed DOI PMC

Fritzsch B., Duncan J. S., Kersigo J., Gray B., Elliott K. L. (2016). Neuroanatomical Tracing Techniques in the Ear: History, State of the Art, and Future Developments. Springer, 243–262. 10.1007/978-1-4939-3615-1_14 PubMed DOI PMC

Fritzsch B., Elliott K. L. (2017). Gene, Cell, and Organ Multiplication Drives Inner Ear Evolution. Dev. Biol. 431, 3–15. 10.1016/j.ydbio.2017.08.034 PubMed DOI PMC

Fritzsch B., Pan N., Jahan I., Duncan J. S., Kopecky B. J., Elliott K. L., et al. (2013). Evolution and Development of the Tetrapod Auditory System: An Organ of Corti-Centric Perspective. Evol. Dev. 15, 63–79. 10.1111/ede.12015 PubMed DOI PMC

Fritzsch B., Straka H. (2014). Evolution of Vertebrate Mechanosensory Hair Cells and Inner Ears: Toward Identifying Stimuli that Select Mutation Driven Altered Morphologies. J. Comp. Physiol. A. 200, 5–18. 10.1007/s00359-013-0865-z PubMed DOI PMC

Frullanti E., Amabile S., Lolli M. G., Bartolini A., Livide G., Landucci E., et al. (2016). Altered Expression of Neuropeptides in FoxG1-Null Heterozygous Mutant Mice. Eur. J. Hum. Genet. 24, 252–257. 10.1038/ejhg.2015.79 PubMed DOI PMC

Goebbels S., Bode U., Pieper A., Funfschilling U., Schwab M. H., Nave K.-A. (2005). Cre/loxP-mediated Inactivation of the bHLH Transcription Factor Gene NeuroD/BETA2. Genesis 42, 247–252. 10.1002/gene.20138 PubMed DOI

Goodrich L. V. (2016). Early Development of the Spiral Ganglion, the Primary Auditory Neurons of the Mammalian Cochlea. Springer 1, 11–48. 10.1007/978-1-4939-3031-9_2 DOI

Guo Z., Zhang L., Wu Z., Chen Y., Wang F., Chen G. (2014). In Vivo Direct Reprogramming of Reactive Glial Cells into Functional Neurons After Brain Injury and in an Alzheimer's Disease Model. Cell Stem Cell 14, 188–202. 10.1016/j.stem.2013.12.001 PubMed DOI PMC

Hébert J. M., McConnell S. K. (2000). Targeting of Cre to the Foxg1 (BF-1) Locus Mediates loxP Recombination in the Telencephalon and Other Developing Head Structures. Dev. Biol. 222, 296–306. 10.1006/dbio.2000.9732 PubMed DOI

Hevner R. F., Hodge R. D., Daza R. A. M., Englund C. (2006). Transcription Factors in Glutamatergic Neurogenesis: Conserved Programs in Neocortex, Cerebellum, and Adult hippocampus. Neurosci. Res. 55, 223–233. 10.1016/j.neures.2006.03.004 PubMed DOI

Huang E. J., Liu W., Fritzsch B., Bianchi L. M., Reichardt L. F., Xiang M. (2001). Brn3a Is a Transcriptional Regulator of Soma Size, Target Field Innervation and Axon Pathfinding of Inner Ear Sensory Neurons. Development 128, 2421–2432. 10.1242/dev.128.13.2421 PubMed DOI PMC

Jahan I., Kersigo J., Pan N., Fritzsch B. (2010). Neurod1 Regulates Survival and Formation of Connections in Mouse Ear and Brain. Cell Tissue Res 341, 95–110. 10.1007/s00441-010-0984-6 PubMed DOI PMC

Jahan I., Pan N., Kersigo J., Fritzsch B. (2010). Neurod1 Suppresses Hair Cell Differentiation in Ear Ganglia and Regulates Hair Cell Subtype Development in the Cochlea. PLoS One 5, e11661. 10.1371/journal.pone.0011661 PubMed DOI PMC

Kasberg A. D., Brunskill E. W., Steven Potter S. (2013). SP8 Regulates Signaling Centers During Craniofacial Development. Dev. Biol. 381, 312–323. 10.1016/j.ydbio.2013.07.007 PubMed DOI PMC

Kawaguchi D., Sahara S., Zembrzycki A., O’Leary D. D. M. (2016). Generation and Analysis of an Improved Foxg1-IRES-Cre Driver Mouse Line. Dev. Biol. 412, 139–147. 10.1016/j.ydbio.2016.02.011 PubMed DOI PMC

Khan S., Chang R. (2013). Anatomy of the Vestibular System: A Review. Nre 32, 437–443. 10.3233/nre-130866 PubMed DOI

Kim W. Y., Fritzsch B., Serls A., Bakel L. A., Huang E. J., Reichardt L. F., et al. (2001). NeuroD-null Mice Are Deaf Due to a Severe Loss of the Inner Ear Sensory Neurons During Development. Development 128, 417–426. 10.1242/dev.128.3.417 PubMed DOI PMC

Krüger M., Schmid T., Krüger S., Bober E., Braun T. (2006). Functional Redundancy of NSCL-1 and NeuroD during Development of the Petrosal and Vestibulocochlear Ganglia. Eur. J. Neurosci. 24, 1581–1590. 10.1111/j.1460-9568.2006.05051.x PubMed DOI

Liberman M. C. (1991). The Olivocochlear Efferent Bundle and Susceptibility of the Inner Ear to Acoustic Injury. J. Neurophysiol. 65, 123–132. 10.1152/jn.1991.65.1.123 PubMed DOI

Liu M., Pereira F. A., Price S. D., Chu M.-j., Shope C., Himes D., et al. (2000). Essential Role of BETA2/NeuroD1 in Development of the Vestibular and Auditory Systems. Genes Dev. 14, 2839–2854. 10.1101/gad.840500 PubMed DOI PMC

Liu M., Pleasure S. J., Collins A. E., Noebels J. L., Naya F. J., Tsai M.-J., et al. (2000). Loss of BETA2/NeuroD Leads to Malformation of the Dentate Gyrus and Epilepsy. Proc. Natl. Acad. Sci. 97, 865–870. 10.1073/pnas.97.2.865 PubMed DOI PMC

Ma L., Harada T., Harada C., Romero M., Hebert J. M., McConnell S. K., et al. (2002). Neurotrophin-3 Is Required for Appropriate Establishment of Thalamocortical Connections. Neuron 36, 623–634. 10.1016/s0896-6273(02)01021-8 PubMed DOI

Ma Q., Anderson D. J., Fritzsch B. (2000). Neurogenin 1 Null Mutant Ears Develop Fewer, Morphologically Normal Hair Cells in Smaller Sensory Epithelia Devoid of Innervation. Jaro 1, 129–143. 10.1007/s101620010017 PubMed DOI PMC

Ma Q., Chen Z., Barrantes I. d. B., Luis de la Pompa J., Anderson D. J. (1998). neurogenin1 Is Essential for the Determination of Neuronal Precursors for Proximal Cranial Sensory Ganglia. Neuron 20, 469–482. 10.1016/s0896-6273(00)80988-5 PubMed DOI

Macova I., Pysanenko K., Chumak T., Dvorakova M., Bohuslavova R., Syka J., et al. (2019). Neurod1 Is Essential for the Primary Tonotopic Organization and Related Auditory Information Processing in the Midbrain. J. Neurosci. 39, 984–1004. 10.1523/jneurosci.2557-18.2018 PubMed DOI PMC

Matei V., Pauley S., Kaing S., Rowitch D., Beisel K. W., Morris K., et al. (2005). Smaller Inner Ear Sensory Epithelia in Neurog1 Null Mice Are Related to Earlier Hair Cell Cycle Exit. Dev. Dyn. 234, 633–650. 10.1002/dvdy.20551 PubMed DOI PMC

Matsuda T., Irie T., Katsurabayashi S., Hayashi Y., Nagai T., Hamazaki N., et al. (2019). Pioneer Factor NeuroD1 Rearranges Transcriptional and Epigenetic Profiles to Execute Microglia-Neuron Conversion. Neuron 101, 472–485. 10.1016/j.neuron.2018.12.010 PubMed DOI

Meas S. J., Zhang C.-L., Dabdoub A. (2018). Reprogramming Glia into Neurons in the Peripheral Auditory System as a Solution for Sensorineural Hearing Loss: Lessons from the Central Nervous System. Front. Mol. Neurosci. 11, 77. 10.3389/fnmol.2018.00077 PubMed DOI PMC

Miyata T., Maeda T., Lee J. E. (1999). NeuroD Is Required for Differentiation of the Granule Cells in the Cerebellum and hippocampus. Genes Dev. 13, 1647–1652. 10.1101/gad.13.13.1647 PubMed DOI PMC

Mostafapour S. P., Cochran S. L., Del Puerto N. M., Rubel E. W. (2000). Patterns of Cell Death in Mouse Anteroventral Cochlear Nucleus Neurons after Unilateral Cochlea Removal. J. Comp. Neurol. 426, 561–571. 10.1002/1096-9861(20001030)426::4<561aid-cne5>3.0.co;2-g PubMed DOI

Packard A., Giel-Moloney M., Leiter A., Schwob J. E. (2011). Progenitor Cell Capacity of NeuroD1-Expressing Globose Basal Cells in the Mouse Olfactory Epithelium. J. Comp. Neurol. 519, 3580–3596. 10.1002/cne.22726 PubMed DOI PMC

Panaliappan T. K., Wittmann W., Jidigam V. K., Mercurio S., Bertolini J. A., Sghari S., et al. (2018). Sox2 Is Required for Olfactory Pit Formation and Olfactory Neurogenesis through BMP Restriction and Hes5 Upregulation. Development 145, dev153791. 10.1242/dev.153791 PubMed DOI PMC

Pang Z. P., Yang N., Vierbuchen T., Ostermeier A., Fuentes D. R., Yang T. Q., et al. (2011). Induction of Human Neuronal Cells by Defined Transcription Factors. Nature 476, 220–223. 10.1038/nature10202 PubMed DOI PMC

Pavlinkova G. (2020). Molecular Aspects of the Development and Function of Auditory Neurons. Int. J. Mol. Sci. 22, 131. 10.3390/ijms22010131 PubMed DOI PMC

Pennesi M. E., Cho J.-H., Yang Z., Wu S. H., Zhang J., Wu S. M., et al. (2003). BETA2/NeuroD1 Null Mice: A New Model for Transcription Factor-dependent Photoreceptor Degeneration. J. Neurosci. 23, 453–461. 10.1523/jneurosci.23-02-00453.2003 PubMed DOI PMC

Romer A. I., Singer R. A., Sui L., Egli D., Sussel L. (2019). Murine Perinatal β-Cell Proliferation and the Differentiation of Human Stem Cell-Derived Insulin-Expressing Cells Require NEUROD1. Diabetes 68, 2259–2271. 10.2337/db19-0117 PubMed DOI PMC

Rubel E. W., Fritzsch B. (2002). Auditory System Development: Primary Auditory Neurons and Their Targets. Annu. Rev. Neurosci. 25, 51–101. 10.1146/annurev.neuro.25.112701.142849 PubMed DOI

Schlosser G., Northcutt R. G. (2000). Development of Neurogenic Placodes inXenopus Laevis. J. Comp. Neurol. 418, 121–146. 10.1002/(sici)1096-9861(20000306)418::2<121aid-cne1>3.0.co;2-m PubMed DOI

Schmidt H., Fritzsch B. (2019). Npr2 Null Mutants Show Initial Overshooting Followed by Reduction of Spiral Ganglion Axon Projections Combined with Near-normal Cochleotopic Projection. Cel Tissue Res 378, 15–32. 10.1007/s00441-019-03050-6 PubMed DOI PMC

Schwarzer S., Asokan N., Bludau O., Chae J., Kuscha V., Kaslin J., et al. (2020). Correction: Neurogenesis in the Inner Ear: the Zebrafish Statoacoustic Ganglion Provides New Neurons from a Neurod/Nestin-Positive Progenitor Pool Well into Adulthood. Development 147, 7. 10.1242/dev.191775 PubMed DOI

Shen W., Ba R., Su Y., Ni Y., Chen D., Xie W., et al. (2018). Foxg1 Regulates the Postnatal Development of Cortical Interneurons. Cereb. Cortex 29(4):1547. 10.1093/cercor/bhy051 PubMed DOI PMC

Wu D. K., Kelley M. W. (2012). Molecular Mechanisms of Inner Ear Development. Cold Spring Harbor Perspect. Biol. 4, a008409. 10.1101/cshperspect.a008409 PubMed DOI PMC

Irx3/5 Null Deletion in Mice Blocks Cochlea-Saccule Segregation and Disrupts the Auditory Tonotopic Map

Harmony in the Molecular Orchestra of Hearing: Developmental Mechanisms from the Ear to the Brain

NEUROD1: transcriptional and epigenetic regulator of human and mouse neuronal and endocrine cell lineage programs

NEUROD1 reinforces endocrine cell fate acquisition in pancreatic development

ISL1 is necessary for auditory neuron development and contributes toward tonotopic organization