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USH2A is a Meissner’s corpuscle protein necessary for normal vibration sensing in mice and humans

Abstract

Fingertip mechanoreceptors comprise sensory neuron endings together with specialized skin cells that form the end-organ. Exquisitely sensitive, vibration-sensing neurons are associated with Meissner’s corpuscles in the skin. In the present study, we found that USH2A, a transmembrane protein with a very large extracellular domain, was found in terminal Schwann cells within Meissner’s corpuscles. Pathogenic USH2A mutations cause Usher’s syndrome, associated with hearing loss and visual impairment. We show that patients with biallelic pathogenic USH2A mutations also have clear and specific impairments in vibrotactile touch perception, as do mutant mice lacking USH2A. Forepaw rapidly adapting mechanoreceptors innervating Meissner’s corpuscles, recorded from Ush2a−/− mice, showed large reductions in vibration sensitivity. However, the USH2A protein was not found in sensory neurons. Thus, loss of USH2A in corpuscular end-organs reduced mechanoreceptor sensitivity as well as vibration perception. Thus, a tether-like protein is required to facilitate detection of small-amplitude vibrations essential for the perception of fine-grained tactile surfaces.

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Fig. 1: USH2A and Ush2a mutations impair cutaneous vibration perception in humans and mice.
Fig. 2: USH2A is expressed in Meissner’s corpuscles and facilitates their vibrosensitivity.
Fig. 3: USH2A is expressed in some hair follicles and facilitates their vibrosensitivity.
Fig. 4: Pacinian corpuscle vibration sensitivity is unchanged in Ush2a−/− mice.
Fig. 5: D-hair and SAM end-organs do not express USH2A, and they have intact mechanosensitivity in Ush2a−/− mice.
Fig. 6: Impaired forepaw RAM mechanosensitivity underlies 5-Hz vibration perceptual deficits in Ush2a−/− mice.

Data availability

Electrophysiological data were acquired with specialized software packages that are not accessible to nonspecialized users. Thus, the raw data files for this manuscript are available upon request.

Code availability

Codes and scripts used in the manuscript are also available upon request.

References

  1. 1.

    Frenzel, H. et al. A genetic basis for mechanosensory traits in humans. PLoS Biol. 10, e1001318 (2012).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Moshourab, R. et al. Congenital deafness is associated with specific somatosensory deficits in adolescents. Sci. Rep. 7, 4251 (2017).

    PubMed  PubMed Central  Google Scholar 

  3. 3.

    Heidenreich, M. et al. KCNQ4 K+ channels tune mechanoreceptors for normal touch sensation in mouse and man. Nat. Neurosci. 15, 138–145 (2012).

    CAS  Google Scholar 

  4. 4.

    Wu, Z. et al. Mechanosensory hair cells express two molecularly distinct mechanotransduction channels. Nat. Neurosci. 20, 24–33 (2016).

    PubMed  PubMed Central  Google Scholar 

  5. 5.

    Eudy, J. D. et al. Mutation of a gene encoding a protein with extracellular matrix motifs in Usher syndrome type IIa. Science 280, 1753–1757 (1998).

    CAS  PubMed  Google Scholar 

  6. 6.

    van Wijk, E. et al. Identification of 51 novel exons of the Usher syndrome type 2A (USH2A) gene that encode multiple conserved functional domains and that are mutated in patients with Usher syndrome type II. Am. J. Human Genet. 74, 738–744 (2004).

    Google Scholar 

  7. 7.

    Adato, A. et al. Usherin, the defective protein in Usher syndrome type IIA, is likely to be a component of interstereocilia ankle links in the inner ear sensory cells. Hum. Mol. Genet. 14, 3921–3932 (2005).

    CAS  PubMed  Google Scholar 

  8. 8.

    Richardson, G. P. & Petit, C. Hair-bundle links: genetics as the gateway to function. Cold Spring Harbor Perspect. Med. 10, a033142 (2019).

    Google Scholar 

  9. 9.

    Milenkovic, N. et al. A somatosensory circuit for cooling perception in mice. Nat. Neurosci. 17, 1560–1566 (2014).

    CAS  PubMed  Google Scholar 

  10. 10.

    Paricio-Montesinos, R. et al. The sensory coding of warm perception. Neuron 106, 830–841.e3 (2020).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Liu, X. et al. Usherin is required for maintenance of retinal photoreceptors and normal development of cochlear hair cells. Proc. Natl Acad. Sci. USA 104, 4413–4418 (2007).

    CAS  PubMed  Google Scholar 

  12. 12.

    Wetzel, C. et al. Small-molecule inhibition of STOML3 oligomerization reverses pathological mechanical hypersensitivity. Nat. Neurosci. 20, 209–218 (2017).

    CAS  PubMed  Google Scholar 

  13. 13.

    Zou, J. et al. Individual USH2 proteins make distinct contributions to the ankle link complex during development of the mouse cochlear stereociliary bundle. Hum. Mol. Genet. 24, 6944–6957 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Hu, J. & Lewin, G. R. Mechanosensitive currents in the neurites of cultured mouse sensory neurones. J. Physiol. 577, 815–828 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Ranade, S. S. et al. Piezo2 is the major transducer of mechanical forces for touch sensation in mice. Nature 516, 121–125 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. 16.

    Usoskin, D. et al. Unbiased classification of sensory neuron types by large-scale single-cell RNA sequencing. Nat. Neurosci. 18, 145–153 (2015).

    CAS  PubMed  Google Scholar 

  17. 17.

    Abdo, H. et al. Specialized cutaneous Schwann cells initiate pain sensation. Science 365, 695–699 (2019).

    CAS  PubMed  Google Scholar 

  18. 18.

    Muniak, M. A., Ray, S., Hsiao, S. S., Dammann, J. F. & Bensmaia, S. J. The neural coding of stimulus intensity: linking the population response of mechanoreceptive afferents with psychophysical behavior. J. Neurosci. 27, 11687–11699 (2007).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Johnson, K. O. The roles and functions of cutaneous mechanoreceptors. Curr. Opin. Neurobiol. 11, 455–461 (2001).

    CAS  PubMed  Google Scholar 

  20. 20.

    Walcher, J. et al. Specialized mechanoreceptor systems in rodent glabrous skin. J. Physiol. 596, 4995–5016 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  21. 21.

    Milenkovic, N., Wetzel, C., Moshourab, R. & Lewin, G. R. Speed and temperature dependences of mechanotransduction in afferent fibers recorded from the mouse saphenous nerve. J. Neurophysiol. 100, 2771–2783 (2008).

    PubMed  Google Scholar 

  22. 22.

    Lechner, S. G. & Lewin, G. R. Hairy sensation. Physiology 28, 142–150 (2013).

    CAS  PubMed  Google Scholar 

  23. 23.

    Li, L. & Ginty, D. D. The structure and organization of lanceolate mechanosensory complexes at mouse hair follicles. eLife 3, e01901 (2014).

    PubMed  PubMed Central  Google Scholar 

  24. 24.

    Joost, S. et al. Single-cell transcriptomics reveals that differentiation and spatial signatures shape epidermal and hair follicle heterogeneity. Cell Systems 3, 221–237.e9 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. 25.

    Johansson, R. S., Landström, U. & Lundström, R. Responses of mechanoreceptive afferent units in the glabrous skin of the human hand to sinusoidal skin displacements. Brain Res. 244, 17–25 (1982).

    CAS  PubMed  Google Scholar 

  26. 26.

    Fleming, M. S. et al. A RET-ER81-NRG1 signaling pathway drives the development of pacinian corpuscles. J. Neurosci. 36, 10337–10355 (2016).

  27. 27.

    Wende, H. et al. The transcription factor c-Maf controls touch receptor development and function. Science 335, 1373–1376 (2012).

    CAS  PubMed  Google Scholar 

  28. 28.

    Prsa, M., Morandell, K., Cuenu, G. & Huber, D. Feature-selective encoding of substrate vibrations in the forelimb somatosensory cortex. Nature 567, 384–388 (2019).

    CAS  PubMed  Google Scholar 

  29. 29.

    Bolanowski, S. J. & Zwislocki, J. J. Intensity and frequency characteristics of pacinian corpuscles. I. Action potentials. J. Neurophysiol. 51, 793–811 (1984).

    PubMed  Google Scholar 

  30. 30.

    Johansson, R. S. & Vallbo, A. B. Detection of tactile stimuli. Thresholds of afferent units related to psychophysical thresholds in the human hand. J. Physiol. 297, 405–422 (1979).

    CAS  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Mountcastle, V. B., Talbot, W. H., Darian-Smith, I. & Kornhubert, H. H. Neural basis of the sense of flutter-vibration. Science 155, 597–600 (1967).

    CAS  PubMed  Google Scholar 

  32. 32.

    Johansson, R. S., Landström, U. & Lundström, R. Sensitivity to edges of mechanoreceptive afferent units innervating the glabrous skin of the human hand. Brain Res. 244, 27–32 (1982).

    CAS  PubMed  Google Scholar 

  33. 33.

    Maksimovic, S. et al. Epidermal Merkel cells are mechanosensory cells that tune mammalian touch receptors. Nature 509, 617–621 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  34. 34.

    Hoffman, B. U. et al. Merkel cells activate sensory neural pathways through adrenergic synapses. Neuron 100, 1401–1413 (2018).

  35. 35.

    Woo, S.-H. et al. Piezo2 is required for Merkel-cell mechanotransduction. Nature 509, 622–626 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Chiang, L.-Y. et al. Laminin-332 coordinates mechanotransduction and growth cone bifurcation in sensory neurons. Nat. Neurosci. 14, 993–1000 (2011).

    CAS  PubMed  Google Scholar 

  37. 37.

    Hu, J., Chiang, L.-Y., Koch, M. & Lewin, G. R. Evidence for a protein tether involved in somatic touch. EMBO J. 29, 855–867 (2010).

    CAS  PubMed  PubMed Central  Google Scholar 

  38. 38.

    Chung, Y. D., Zhu, J., Han, Y. & Kernan, M. J. nompA encodes a PNS-specific, ZP domain protein required to connect mechanosensory dendrites to sensory structures. Neuron 29, 415–428 (2001).

    CAS  PubMed  Google Scholar 

  39. 39.

    Poole, K., Herget, R., Lapatsina, L., Ngo, H.-D. & Lewin, G. R. Tuning Piezo ion channels to detect molecular-scale movements relevant for fine touch. Nat. Commun. 5, 3520 (2014).

    PubMed  PubMed Central  Google Scholar 

  40. 40.

    Fettiplace, R. & Kim, K. X. The physiology of mechanoelectrical transduction channels in hearing. Physiol. Rev. 94, 951–986 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  41. 41.

    Murthy, S. E. et al. The mechanosensitive ion channel Piezo2 mediates sensitivity to mechanical pain in mice. Sci. Trans. Med. 10, eaat9897 (2018).

  42. 42.

    Chesler, A. T. et al. The role of PIEZO2 in human mechanosensation. N. Engl. J. Med. 375, 1355–1364 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  43. 43.

    Kazmierczak, P. et al. Cadherin 23 and protocadherin 15 interact to form tip-link filaments in sensory hair cells. Nature 449, 87–91 (2007).

    CAS  PubMed  Google Scholar 

  44. 44.

    Rolke, R. et al. Quantitative sensory testing in the German Research Network on Neuropathic Pain (DFNS): standardized protocol and reference values. Pain 123, 231–243 (2006).

    CAS  PubMed  Google Scholar 

  45. 45.

    Geyer, M. A. & Dulawa, S. C. Assessment of murine startle reactivity, prepulse inhibition, and habituation. Curr. Protoc. Neurosci. Chapter 8, Unit 8 (17) (2003).

    Google Scholar 

  46. 46.

    Chaplan, S. R., Bach, F. W., Pogrel, J. W., Chung, J. M. & Yaksh, T. L. Quantitative assessment of tactile allodynia in the rat paw. J. Neurosci. Methods 53, 55–63 (1994).

    CAS  PubMed  Google Scholar 

  47. 47.

    Carandini, M. & Churchland, A. K. Probing perceptual decisions in rodents. Nat. Neurosci. 16, 824–831 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  48. 48.

    Macmillan, N. A. & Kaplan, H. L. Detection theory analysis of group data: estimating sensitivity from average hit and false-alarm rates. Psychol. Bull. 98, 185–199 (1985).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Braunschweig and H. Thränhardt for technical assistance, T. Li (National Institutes of Health) for providing us with Ush2a−/− mice, B. Purfürst for electron microscopy, M. Navarro for assistance with patient recruitment and A. Udhayachandran for advice. The present study was funded by grants from the Deutsche Forschungsgemeinshaft (grant nos. SFB665-B6 to G.R.L., SFB1315 to J.F.A.P. and SFB1158-A01 to S.G.L.) and grants from the European Research Council (grant nos. 789128 to G.R.L. and ERC-2015-CoG-682422 to J.F.A.P.). Additional funding was from the Institute of Health Carlos III (Spanish Ministry of Science and Innovation, grant no. FIS PI16/00539 to J.M.). We thank U. Müller for critical comments on the manuscript.

Author information

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Authors

Contributions

F.S., V.B., R.M. and G.R.L. conceived the human and mouse studies. F.S. carried out mouse skin electrophysiology and behavior experiments with guidance and help from R.P.M., G.R.L. and J.F.A.P. F.J.T. and S.G.L. established, performed and analyzed data from Pacinian afferent recordings. F.S., B.M. and T.D. carried out immunohistochemical and anatomical experiments. V.B. and R.M. recruited the control cohort, and collected and analyzed the quantitative sensory testing data of the control cohort. Psychophysical experiments with Spanish patients with Usher’s syndrome, recruited by J.M.M. and G.G.-G., were carried out and organized by V.B., F.S., G.G.-G., J.M.M., J.K. and J.O.A. F.S. and G.R.L. wrote the manuscript with input from all the authors.

Corresponding author

Correspondence to Gary R. Lewin.

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The authors declare no competing interests.

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Peer review information Nature Neuroscience thanks Victoria Abraira, Daniel Huber and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Mouse vibration perceptual training task.

a, Hit and False alarm lick rates for Ush2a+/+ and b, Ush2a-/- mice during training. Ush2a+/+ and Ush2a-/- mice learned to report 5Hz 60mN forepaw vibration after 4 days (Hit vs False lick rates within each genotype: ***P < 0.001; two-way repeated-measures ANOVA with Bonferroni post-hoc analysis. Ush2a+/+: F(6,60) = 4.486, P=0.006. Ush2a-/-: F(6,48) = 3.698,P=0.004). Light colour lines indicate individual mice, bold lines population mean. c, Perceptual sensitivity (d’) measurements indicate increasing sensitivity during training in both Ush2a+/+ and Ush2a-/- mice. Plot shows max and min values, box shows 1st, 2nd (median) and 3rd quartile values. d, Perceptual thresholds of Ush2a+/+ mice to 5Hz stimuli. Ush2a+/+ mice reported 5Hz stimuli ≥12mN, as shown by significantly higher Hit vs False Alarm lick rates ≥12mN (Hit vs False: Two-way repeated measures ANOVA F(5,30) = 16.52, P=0.0005, asterisk indicates Bonferroni post-hoc analyses ***P < 0.001. e, Ush2a-/- mice reported 5Hz vibration stimuli ≥24mN (Hit vs False: two-way repeated-measures ANOVA F(6,24) = 16.52, P<0.0001, asterisk indicates Bonferroni post-hoc analyses ***P < 0.001. f, Mean first lick latencies during hit trials of Ush2a-/- mice were longer at 36mN 5Hz vibration; Two-way repeated-measures ANOVA F(6,63) = 2.306, P=0.045 asterisk indicates Bonferroni post-hoc analysis *P < 0.05. g, Hit and False licking rates during 25Hz vibration detection task in Ush2a+/+ and Ush2a-/- mice. Dots show data from individual mice, horizontal lines show mean. h, Hit and False licking rates of mice during 100Hz vibration detection task in Ush2a+/+ and Ush2a-/- mice. i, Mice trained on the vibration detection did not respond to any sound generated by the vibration stimulus when the Piezo device placed 5mm below the forepaw (paired two-tailed t-tests, Ush2a+/+ P=0.97; Ush2a-/- P=0.17). Data = mean ± s.e.m. n = 11 mice. Filled dot shows hit rates, open dot shows false alarm lick rates.

Extended Data Fig. 2 Ush2a-/- mice have a mild hearing deficit in a paired-pulse inhibition behavioral task.

a, Response amplitudes (in arbitrary units) of Ush2a+/+ (n=10) and Ush2a-/- (n=9) mice to 20-40ms auditory tones of different amplitudes did not differ (Two-way repeated-measures ANOVA analysis F(1,18)=2.937, P=0.10). b, Lower amplitude tones reduced the startle response when given 200ms before the startling 129dB tone in both Ush2a+/+ (n=10) and Ush2a-/- (n=9) mice, however the amplitude of the response was significantly lower in Ush2a-/- mice compared to Ush2a+/+ mice at 73dB and 81bD pre-pulse tones; two-way repeated-measures ANOVA F(1,18)=2.711, P=0.006, asterisks indicate Bonferroni post-hoc analyses *P < 0.01. Data = mean ± s.e.m.

Extended Data Fig. 3 USH2A immunolabelling in cutaneous Meissner corpuscles.

a, USH2A (green) and NF200 (red) immunolabelling of dorsal root ganglia neurons. a’ USH2A channel only. a’’ NF200 channel only. A’’’ RNA scope probe for Ush2a mRNA shows no labelling in the DRG (green) a second probe for NF200 mRNA (red) shows large sensory neurons labeled. The Ush2a RNA labels Meissner corpuscle cells in the skin (green). b NF200 (red), and S100 (cyan) immunolabelling of Meissner corpuscles in the sectioned paw glabrous skin. b’ USH2A channel. b’’ NF200 channel. b’’’ S100 . c, USH2A (green) and NF200 (red) immunolabelling of deep nerve branches in whole mounted paw skin show an absence of USH2A in nerve bundles. c’ USH2A channel only. c’’ NF200 channel. d, USH2A (green) and NF200 (red) immunolabelling of Meissner corpuscles in the sectioned paw glabrous skin from an Ush2a-/- mouse show an absence of USH2A labelling. d’ USH2A channel only. d’’ NF200 channel only.

Extended Data Fig. 4 Ex vivo forepaw and hind paw skin nerve preparation recordings from cutaneous RAM Aβ-fibers.

a, The proportion of forepaw RAMs responding to 5Hz (top) and 50Hz (bottom) vibration stimuli in Ush2a+/+ (n = 11 from 5 mice) and Ush2a-/- mice (n = 10 from 4 mice). b, Ush2a-/- forepaw RAMs (n = 10) were less responsive to 10Hz, two-way repeated-measures ANOVA F(1,12) = 8.277, P=0.014 and c, 50Hz vibration, two-way repeated-measures ANOVA F(1,19) = 8.155, P=0.010 compared to RAMs in Ush2a+/+ mice (n = 11) asterisks indicate Bonferroni post-hoc analyses **P < 0.01, *** P < 0.001. d, Number of spikes per sinusoid decreases with stimulus amplitude in Ush2a+/+ forepaw RAMs (n = 11) compared to Ush2a-/- (n = 10). e. Number of spikes per sinusoid for vibration stimuli (5-50 Hz) was lower in Ush2a-/- forepaw RAMs (n = 10) compared to Ush2a+/+ (n = 11). f, Hind paw glabrous Aβ-fiber RAMs recorded from Ush2a-/- mice (n = 14 from 4 mice) were less responsive to 5Hz, two-way repeated-measures ANOVA F(1,25) = 8.028, P=0.009) (left asterisks indicate Bonferroni post-hoc analyses *,**P < 0.05, 0.01. g, but not 50Hz vibration compared to Ush2a+/+ mice (n = 13 from 4 mice), two-way repeated-measures ANOVA F(1,24) = 2.101, P=0.16. h, Hind paw glabrous RAM responses to different ramp velocities were significantly lower in Ush2a-/- mice (n = 10) compared to Ush2a+/+ mice (n = 11), two-way repeated-measures ANOVA. F(1,43) = 8.112, P=0.007, asterisks indicate Bonferroni post-hoc analyses *P < 0.05. i, Forepaw Aβ-fiber RAMs (n = 11 neurons from 5 mice) were significantly more sensitive to 5Hz and, j, 50Hz vibration compared to hind paw RAMs in Ush2a+/+ mice two-way repeated-measures ANOVA. 5Hz: F(1,21)=40.26, P<0.0001; 50Hz: F(1,16)=6.970, P=0.018; asterisks indicate Bonferroni post-hoc analyses *,**,***P < 0.05, 0.01, 0.001 (n = 13, 4 mice). k, Aβ-fiber SAM responses to 5Hz vibration were significantly higher in the forepaw (n = 12) compared to hindpaw (n = 14) in Ush2a+/+ mice, two-way repeated-measures ANOVA F(1,21) = 8.210, P=0.009) Data = mean ± s.e.m.

Extended Data Fig. 5 USH2A immunolabelling in the mouse skin.

a, Whole mount USH2A (green) and S100 immunolabelling of a hair follicle with lanceolate endings in the hind paw hairy skin. a’ USH2A channel only. a’’ S100 channel only. Scale bars 25µm. b, Wholemount USH2A (green) and S100 (red) immunolabelling of a hair follicle with circumferential (circ) endings showing USH2A expression in the hind paw hairy skin. b’ USH2A channel only. b’’ S100 channel only. Scale bars 25µm. c, Wholemount USH2A (green) and S100 (red) immunolabelling of a hair follicle with circumferential endings that is USH2A-negative in the hind paw hairy skin. c’ USH2A channel only. c’’ NF200 channel only. Scale bars 25µm. d, Whole mount USH2A (green) and CK20 (red: marker of Merkel cells) immunolabelling of Guard hair in the back hairy skin with surrounding Merkel cell complex. Note the lack of overlap between USH2A and CK20 immunolabelling. d’ USH2A channel only. d’’ S100 channel only. Scale bars 50µm. e, USH2A (green) and S100 (red) immunolabelling of D-hairs in sectioned glabrous hind paw skin, showing absence of USH2A staining in D-hairs. e’ USH2A channel only. e’’ S100 channel only. All scale bars = 50µm.

Extended Data Fig. 6 Aβ-fiber SAMs and Aδ-fiber D-hairs have normal mechanosensitivity in Ush2a-/- mice.

a, Example traces of single SAMs recorded from Ush2a+/+ (top) and Ush2a-/- (bottom) mice firing in response to the 5Hz vibration protocol. b, SAMs recorded from Ush2a+/+ (n=10 from 4 mice) and Ush2a-/- (n=11 from 4 mice) mice show comparable sensitivity to 25Hz and c, 50Hz vibration (Ush2a+/+ n=10; Ush2a-/- n=11); Two-way repeated-measures ANOVA F(1,19)=0.3011, P=0.59. d, Aδ-fiber D-hairs recorded from the hairy hind paw skin and glabrous hind paw skin in Ush2a+/+ (n=9 from 6 mice) and Ush2a-/- (n=11 from 6 mice) mice showed comparable sensitivity to 25Hz and e, 50Hz vibration (Two-way repeated-measures ANOVA 25Hz: F (1,19)=0.00006, P=0.99. 50Hz: F(1,19) > 0.1, P=0.68). f, D-hair vibration frequency tuning was not significantly different in Ush2a+/+ and Ush2a-/- mice (Ush2a+/+ n=9; Ush2a-/- n=11. Two-way repeated-measures ANOVA F(1,17) = 0.0001, P=0.99). All data = mean ± s.e.m.

Extended Data Fig. 7 Warm and cool detection by forepaw thermoreceptors is not impaired in Ush2a-/- mice.

a, The percentages of forepaw thermoreceptive C- and A-fibers classified by their response to different stimuli in Ush2a+/+ and Ush2a-/- mice (n=48 from 6 mice). C-MH = C-mechanoheat; C-MHC = C-mechanoheatcold; C-MC = C-mechanocold; C-C = C-cold; A-MC = A-mechanocold. b, Firing rates of cool-sensitive afferents recorded in Ush2a+/+ and Ush2a-/- mice during a 32-12 °C ramp (temperature change = 1 °C/second) was not statistically different (two-way repeated-measures ANOVA F(1,31)=0.0013, P=0.97). c, Similarly, firing rates of heat-sensitive thermoreceptors during a 32-48 °C ramp did not differ between Ush2a+/+ and Ush2a-/- mice (two-way repeated-measures ANOVA F(1,20)=0.2716, P=0.61). Data = mean ± s.e.m.

Extended Data Fig. 8 Tibial nerve sensory neurons and hind paw skin touch end-organs in Ush2a-/- mice show no anatomical defects.

a, electron micrographs of the tibial nerve from an Ush2a+/+ mouse. Myelinated fibers have thick grey circumference; Remak bundles of C-fibers lack myelination. a’, Representative electron micrograph of the tibial nerve from an Ush2a-/- mouse. Scale bars = 2µm. b, Mean numbers of A and C-fibers in the tibial nerves of Ush2a+/+ (n = 3) and Ush2a-/- mice (n = 3) did not appear to differ. Data quantified from EM images. c, Myelin thickness of tibial axons did not differ between Ush2a+/+ (n = 3) and Ush2a-/- mice (n = 3). d, Example S100 (green) and NF200 (red) labelling of Meissner corpuscles in the glabrous hind paw skin of an Ush2a+/+ mouse and e, an Ush2a-/- mouse showed no morphological deficits. Scale bars = 50µm. f, Mean number of fibers innervating each Meissner corpuscle did not differ between Ush2a+/+ (n = 5 mice) and Ush2a-/- mice (n = 5 mice: P < 0.05; two-way ANOVA, F(1,16)=0.0, P=0.99), whisker indicates max and min values, box indicates 1st 2nd (median) and 3rd quartiles. g, Immunolabelling S100 (green) and NF200 (red) labels hair follicles in the skin of an Ush2a+/+ mouse. Scale bar = 25µm. h, S100 (green) and NF200 (red) labelling hair follicles in the skin of an Ush2a-/- mouse showed no obvious deficits. Scale bar = 25µm. i, Hair bulb thickness of awl/zigzag hairs and Guard hairs did not differ between Ush2a+/+ (n = 5) and Ush2a-/- mice (n = 5), unpaired two-tailed t-tests, P=0.64 and P=0.77. Similarly, j, number of terminal Schwann cells per hair follicle (Ush2a+/+ n = 5, Ush2a-/- mice n = 5. P=0.30 and P=0.93.) k, mean terminal Schwann cell soma size (Ush2a+/+ n = 5, Ush2a-/- mice n = 5. P=0.57 and P=0.28) l, and branch lengths did not differ between Ush2a+/+ (n = 5) and Ush2a-/- mice (n = 5), unpaired two-tailed t-tests, P=0.65 and P=0.39). Data = mean ± s.e.m.

Extended Data Fig. 9 Impaired forepaw RAM mechanosensitivity underlies 25Hz vibration perceptual deficits in Ush2a-/- mice.

a, Box and whisker plot of median first lick latencies of Ush2a+/+ (n=6) and Ush2a-/- (n=5) mice at 5Hz, 25Hz and 100Hz frequency vibration, whisker indicates max and min values, box indicates 1st 2nd (median) and 3rd quartiles. b PSTHs of first lick responses to 25Hz 12mN vibration stimuli from all behaving Ush2a+/+ and Ush2a-/- trials. Peak first lick Hit responses above False lick rates indicate first responses of mice 25Hz stimuli. Ush2a-/- mice did not detect this stimulus c Spike timing raster plots of individual forepaw RAMs and SAMs recorded in response to the first 20 cycles of a 25Hz vibration stimulus in the ex vivo forepaw skin nerve preparation of Ush2a+/+ and Ush2a-/- mice. Each row represents one trial from a single afferent 1-3 trials per recording (left Ush2a+/+ RAM n = 4/11, SAM 2/10 afferents; right Ush2a-/- RAM n = 1/10, SAM 1/11 afferents). d, PSTHs of mean first lick responses to 60mN 25Hz vibration stimuli from all behaving Ush2a+/+ and Ush2a-/-, both genotypes detected this stimulus. e, Spike timings of individual RAMs and SAMs recorded in the ex vivo forepaw skin nerve preparation in Ush2a+/+ and Ush2a-/- mice in response to the first 20 cycles of 60mN 25Hz stimuli (left Ush2a+/+ RAM n = 10/11, SAM n = 10/10; right Ush2a-/- RAM n = 8/10, SAM n = 10/11). Data = mean ± s.e.m.

Extended Data Fig. 10 Piezo actuator calibration for displacement and force.

a, Force measurement system mounted in series with the Piezo actuator was used to deliver vibration stimuli to the ex vivo preparation. The displacement of the probe was measured and compared with the measured force for the same output voltage when used in the ex vivo preparation. This configuration was used to apply calibrated vibration stimuli to skin mechanoreceptors.

Supplementary information

Supplementary Information

Supplementary Tables 1–3.

Reporting summary

Supplementary Video 1

Animated z-projection of the Meissner’s corpuscle pictured in Fig. 2a.

Supplementary Video 2

Animated z-axis projections of a single Meissner’s corpuscle labeled for USH2A (green), S100B (magenta) and NF200 sensory axons (blue).

Supplementary Video 3

Animated z-axis projections of a single Meissner’s corpuscle labeled for USH2A (green), S100B (magenta) and NF200 sensory axons (blue).

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Schwaller, F., Bégay, V., García-García, G. et al. USH2A is a Meissner’s corpuscle protein necessary for normal vibration sensing in mice and humans. Nat Neurosci 24, 74–81 (2021). https://doi.org/10.1038/s41593-020-00751-y

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