Roles of the Espin Actin-Bundling Proteins in the Morphogenesis and Stabilization of Hair Cell Stereocilia Revealed in CBA/CaJ Congenic Jerker Mice

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Hearing and vestibular function depend on mechanosensory staircase collections of hair cell stereocilia, which are produced from microvillus-like precursors as their parallel actin bundle scaffolds increase in diameter and elongate or shorten. Hair cell stereocilia contain multiple classes of actin-bundling protein, but little is known about what each class contributes. To investigate the roles of the espin class of actin-bundling protein, we used a genetic approach that benefited from a judicious selection of mouse background strain and an examination of the effects of heterozygosity. A congenic jerker mouse line was prepared by repeated backcrossing into the inbred CBA/CaJ strain, which is known for excellent hearing and minimal age-related hearing loss. We compared stereocilia in wild-type CBA/CaJ mice, jerker homozygotes that lack espin proteins owing to a frameshift mutation in the espin gene, and jerker heterozygotes that contain reduced espin levels. The lack of espins radically impaired stereociliary morphogenesis, resulting in stereocilia that were abnormally thin and short, with reduced differential elongation to form a staircase. Mean stereociliary diameter did not increase beyond ∼0.10–0.14 µm, making stereocilia ∼30%–60% thinner than wild type and suggesting that they contained ∼50%–85% fewer actin filaments. These characteristics indicate a requirement for espins in the appositional growth and differential elongation of the stereociliary parallel actin bundle and fit the known biological activities of espins in vitro and in transfected cells. The stereocilia of jerker heterozygotes showed a transient proximal-distal tapering suggestive of haploinsufficiency and a slowing of morphogenesis that revealed previously unrecognized assembly steps and intermediates. The lack of espins also led to a region-dependent degeneration of stereocilia involving shortening and collapse. We conclude that the espin actin-bundling proteins are required for the assembly and stabilization of the stereociliary parallel actin bundle.

Vyšlo v časopise: Roles of the Espin Actin-Bundling Proteins in the Morphogenesis and Stabilization of Hair Cell Stereocilia Revealed in CBA/CaJ Congenic Jerker Mice. PLoS Genet 7(3): e32767. doi:10.1371/journal.pgen.1002032
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002032

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1. VollrathMAKwanKYCoreyDP 2007 The micromachinery of mechanotransduction in hair cells. Annu Rev Neurosci 30 339 365

2. SchwanderMKacharBMüllerU 2010 The cell biology of hearing. J Cell Biol 190 9 20

3. BartlesJR 2000 Parallel actin bundles and their multiple actin-bundling proteins. Curr Opin Cell Biol 12 72 78

4. TilneyLGTilneyMSDeRosierDJ 1992 Actin filaments, stereocilia and hair cells: how cells count and measure. Annu Rev Cell Biol 8 257 274

5. FrolenkovGIBelyantsevaIAFriedmanTBGriffithAJ 2004 Genetic insights into the morphogenesis of inner ear hair cells. Nat Rev Genet 5 489 498

6. TilneyLGDeRosierDJ 1986 Actin filaments, stereocilia, and hair cells of the bird cochlea. IV. How the actin filaments become organized in developing stereocilia and in the cuticular plate. Dev Biol 116 119 129

7. TilneyLGSaundersJC 1983 Actin filaments, stereocilia, and hair cells of the bird cochlea. I. Length, number, width, and distribution of stereocilia of each hair cell are related to the position of the hair cell on the cochlea. J Cell Biol 96 807 821

8. TilneyLGTilneyMSCotancheDA 1988 Actin filaments, stereocilia, and hair cells of the bird cochlea. V. How the staircase pattern of stereociliary lengths is generated. J Cell Biol 106 355 365

9. KaltenbachJAFalzaranoPRSimpsonTH 1994 Postnatal development of the hamster cochlea: II. Growth and differentiation of stereocilia bundles. J Comp Neurol 350 197 198

10. LiAXueJPetersonEH 2008 Architecture of the mouse utricle: macular organization and hair bundle heights. J Neurophysiol 99 718 733

11. DrorAAAvrahamKB 2009 Hearing loss: mechanisms revealed by genetics and cell biology. Annu Rev Genet 43 411 437

12. PetitCRichardsonGP 2009 Linking genes underlying deafness to hair-bundle development and function. Nat Neurosci 12 703 710

13. TilneyMSTilneyLGStephensREMerteCDrenckhahnD 1989 Preliminary characterization of the stereocilia and cuticular plate of hair cells in the chick cochlea. J Cell Biol 109 1711 1723

14. ZhengLSekerkováGVranichKTilneyLGMugnainiE 2000 The deaf jerker mouse has a mutation in the gene encoding the espin actin-bundling proteins of hair cell stereocilia and lacks espins. Cell 102 377 385

15. DaudetNLebartM-C 2002 Transient expression of the T-isoform of plastins/fimbrin in the stereocilia of developing auditory hair cells. Cell Motil Cytoskeleton 53 326 336

16. KitajiriSSakamotoTBelyantsevaIAGoodyearRJStepanyanR 2010 Actin-bundling protein TRIOBP forms resilient rootlets of hair cell stereocilia essential for hearing. Cell 141 786 798

17. ShinJBLongo-GuessCMGagnonLHSaylorKWDumontRA 2010 The R109H variant of fascin-2, a developmentally regulated actin crosslinker in hair-cell stereocilia, underlies early-onset hearing loss of DBA/2J mice. J Neurosci 30 9683 9694

18. PengAWBelyantsevaIAHsuPDFriedmanTBHellerS 2009 Twinfilin 2 regulates actin filament lengths in cochlear stereocilia. J Neurosci 29 15083 15088

19. RzadzinskaAKNevalainenNKProsserHMLappalainenPSteelKP 2009 Myosin VIIA interacts with twinfilin-2 at the tips of mechanosensory stereocilia in the inner ear. PLoS ONE 4 e7097 doi:10.1371/journal.pone.0007097

20. MburuPRomeroHRHiltonHParkerATownsendS 2010 Gelsolin plays a role in the actin polymerization complex of hair cell stereocilia. PLoS ONE 5 e11627 doi:10.1371/journal.pone.0011627

21. BelyantsevaIABogerETNazSFrolenkovGISellersJR 2005 Myosin-XVa is required for tip localization of whirlin and differential elongation of hair-cell stereocilia. Nat Cell Biol 7 148 156

22. DelpratBMichelVGoodyearRYamasakiYMichalskiN 2005 Myosin XVa and whirlin, two deafness gene products required for hair bundle growth, are located at stereocilia tips and interact directly. Hum Mol Genet 14 401 410

23. SallesFTMerrittRCJrManorUDohertyGWSousaAD 2009 Myosin IIIa boosts elongation of stereocilia by transporting espin 1 to the plus ends of actin filaments. Nat Cell Biol 11 443 450

24. SekerkováGZhengLLoomisPAMugnainiEBartlesJR 2006 Espins and the actin cytoskeleton of hair cell stereocilia and sensory cell microvilli. Cell Mol Life Sci 63 2329 241

25. BartlesJRWierdaAZhengL 1996 Identification and characterization of espin, an actin-binding protein localized to the F-actin-rich junctional plaque of Sertoli cell ectoplasmic specializations. J Cell Sci 110 1229 1239

26. SekerkováGLoomisPAChangyaleketBZhengLEytanR 2003 Novel espin actin-bundling proteins are localized to Purkinje cell dendritic spines and bind the Src homology 3 adapter protein insulin receptor substrate p53. J Neurosci 23 1310 1319

27. SekerkováGZhengLLoomisPAChangyaleketBWhitlonDS 2004 Espins are multifunctional actin cytoskeletal regulatory proteins in the microvilli of chemosensory and mechanosensory cells. J Neurosci 24 5445 5456

28. BartlesJRZhengLLiAWierdaAChenB 1998 Small espin: a third actin-bundling protein and potential forked protein ortholog in brush border microvilli. J Cell Biol 143 107 119

29. ChenBLiAWangDWangMZhengL 1999 Espin contains an additional actin-binding site in its N terminus and is a major actin-bundling protein of the Sertoli cell-spermatid ectoplasmic specialization junctional plaque. Mol Biol Cell 10 4327 4339

30. PurdyKRBartlesJRWongGC 2007 Structural polymorphism of the actin-espin system: a prototypical system of filaments and linkers in stereocilia. Phys Rev Lett 98 08105

31. ShinHPurdy DrewKRBartlesJRWongGCLGrasonGM 2010 Cooperativity and frustration in protein-mediated parallel actin bundles. Phys Rev Lett 103 238102

32. LoomisPAZhengLSekerkováGChangyaleketBMugnainiE 2003 Espin cross-links cause the elongation of microvillus-type parallel actin bundles in vivo. J Cell Biol 163 1045 1055

33. RzadzinskaASchneiderMNoben-TrauthKBartlesJRKacharB 2005 Balanced levels of espin are critical for stereociliary growth and length maintenance. Cell Motil Cytoskeleton 62 157 165

34. ZhengLZhengJWhitlonDSGarcía-AñoverosJBartlesJR 2010 Targeting of the hair cell proteins cadherin 23, harmonin, myosin XVa, espin and prestin in an epithelial cell model. J Neurosci 30 7187 7201

35. SekerkováGZhengLMugnainiEBartlesJR 2006 Differential expression of espin isoforms during epithelial morphogenesis, stereociliogenesis and postnatal maturation in the developing inner ear. Dev Biol 291 83 95

36. FurnessDNMahendrasingamSOhashiMFettiplaceRHackneyCM 2008 The dimensions and composition of stereociliary rootlets in mammalian cochlear hair cells: comparison between high- and low-frequency cells and evidence for a connection to the lateral membrane. J Neurosci 28 6342 6353

37. LiHLiuHBaltSMannSCorralesCE 2004 Correlation of expression of the actin filament-bundling protein espin with stereociliary bundle formation in the developing inner ear. J Comp Neurol 468 125 134

38. OshimaKShinKDiensthuberMPengAWRicciAJ 2010 Mechanosensitive hair cell-like cells from embryonic and induced pluripotent stem cells. Cell 141 704 716

39. NazSGriffithAJRiazuddinSHamptonLLBatteyJFJr 2004 Mutations of ESPN cause autosomal recessive deafness and vestibular dysfunction. J Med Genet 41 591 595

40. DonaudyFZhengLFicarellaRBallanaECarellaM 2006 Espin gene (ESPN) mutations associated with autosomal dominant hearing loss cause defects in microvillar elongation or organization. J Med Genet 43 157 161

41. GrünebergHBurnettJBSnellGD 1941 The origin of jerker, a new gene mutation of the house mouse, and linkage studies made with it. Proc Natl Acad Sci USA 27 562 565

42. DeolMS 1954 The anomalies of the labyrinth of the mutants varitint-waddler, shaker-2 and jerker in the mouse. J Genet 52 562 565

43. SteelKPBockGR 1983 Cochlear dysfunction in the jerker mouse. Behav Neurosci 97 381 391

44. JacksonTThomasJGreenEDNoben-TrauthK 2002 Genetic and physical maps of jerker (Espnje) on mouse chromosome 4. Biochem Biophys Res Commun 296 1143 1147

45. SjöströmBAnnikoM 1990 Morphologically specific vestibular hair cell degeneration in the jerker mutant mouse. Eur Arch Otorhinolaryngol 247 51 55

46. SjöströmBAnnikoM 1992 Cochlear structure and function in a recessive type of genetically induced inner ear degeneration. ORL J Otorhinolaryngol Relat Spec 54 220 228

47. SjöströmBAnnikoM 1992 Genetically induced inner ear degeneration. A structural and functional study. Act Otolaryngol Suppl (Stockh) 493 141 146

48. SjöströmBAnnikoM 1990 Variability in genetically induced age-related impairment of auditory brainstem response thresholds. Acta Otolaryngol 109 353 360

49. JohnsonKRZhengQYNoben-TrauthK 2006 Strain background effects and genetic modifiers of hearing in mice. Brain Res 1091 79 88

50. ZhengQYJohnsonKRErwayLC 1999 Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses. Hear Res 130 94 107

51. SekerkováGZhengLMugnainiEBartlesJR 2008 Espin actin-cytoskeletal proteins are in rat type I spiral ganglion neurons and include splice-isoforms with a functional nuclear localization signal. J Comp Neurol 509 661 676

52. RothBBrunsV 1992 Postnatal development of the rat organ of Corti: II. Hair cell receptors and their supporting elements. Anat Embryol (Berl) 185 571 581

53. ShnersonAWillottJF 1980 Ontogeny of the acoustic startle response in C57BL/6J mouse pups. J Comp Physiol Psychol 94 36 40

54. EtournayRLepelletierLBoutet de MonvelJMichelVCayetN 2010 Cochlear outer hair cells undergo an apical circumference remodeling constrained by the hair bundle shape. Development 137 1373 1383

55. DesaiSSZehCLysakowskiA 2005 Comparative morphology of rodent vestibular periphery. I. Saccular and utricular maculae. J Neurophysiol 93 251 266

56. LelliAAsaiYForgeAHoltJRGéléocGS 2009 Tonotopic gradient in the developmental acquisition of sensory transduction in outer hair cells of the mouse cochlea. J Neurophysiol 101 2961 2973

57. LoomisPAKellyAEZhengLChangyaleketBSekerkováG 2006 Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to actin bundles in cells. J Cell Sci 119 1655 1665

58. ZigmondSHFurukawaRFechheimerM 1992 Inhibition of actin filament depolymerization by the Dictyostelium 30,000-D actin-bundling protein. J Cell Biol 119 559 567

59. SchmollerKMSemmrichCBauschAR 2011 Slow-down of actin depolymerization by cross-linking molecules. J Struct Biol 173 350 357

60. ShinJHMahadevanLSoPTMatsudairaP 2004 Bending stiffness of a crystalline actin bundle. J Mol Biol 337 255 261

61. BelyantsevaIAPerrinBJSonnemannKJZhuMStepanyanR 2009 Gamma-actin is required for cytoskeletal maintenance but not development. Proc Natl Acad Sci USA 106 9703 9708

62. DesaiSSAliHLysakowskiA 2005 Comparative morphology of the rodent vestibular periphery. II. Cristae ampullares. J Neurophysiol 93 267 280

63. SansAChatM 1982 Analysis of temporal and spatial patterns of rat vestibular hair cell differentiation by tritiated thymidine radioautography. J Comp Neurol 206 1 8

64. GoldbergJMDesmadrylGBairdRAFernándezC 1990 The vestibular nerve of the chinchilla. V. Relation between afferent discharge properties and peripheral innervation patterns in the utricular macula. J Neurophysiol 63 791 804

65. WooltortonJRAGaboyardSHurleyKMPriceSDGarciaJL 2007 Developmental changes in two voltage-dependent sodium currents in utricular hair cells. J Neurophysiol 97 1684 1704

66. GoodyearRRichardsonG 1992 Distribution of the 275 kD hair cell antigen and cell surface specialisations on auditory and vestibular hair bundles in the chicken inner ear. J Comp Neurol 325 243 256

67. ZineAHafidiARomandR 1995 Fimbrin expression in the developing rat cochlea. Hear Res 87 165 169

68. VolkmannNDeRosierDMatsudairaPHaneinD 2001 An atomic model of actin filaments cross-linked by fimbrin and its implications for bundle assembly and function. J Cell Biol 153 947 956

69. ClaessensMMSemmrichCRamosLBauschAR 2008 Helical twist controls the thickness of actin bundles. Proc Natl Acad Sci USA 105 8819 8822

70. TilneyLGConnellyPSVranichKAShawMKGuildGM 1998 Why are two different cross-linkers necessary for actin bundle formation in vivo and what does each cross-linker contribute? J Cell Biol 143 121 133

71. Hunter-DuvarIM 1978 A technique for preparation of cochlear specimens for assessment with the electron microscope. Acta Otolaryngol 351 3 23

72. PearceMRichterC-PCheathamMA 2001 A reconsideration of sound calibration in the mouse. J Neurosci Methods 106 57 67