Actin at stereocilia tips is regulated by mechanotransduction and ADF/cofilin

McGrath, Jamis; Tung, Chun-Yu; Liao, Xiayi; Belyantseva, Inna A.; Roy, Pallabi; Chakraborty, Oisorjo; Li, Jinan; Berbari, Nicolas F.; Faaborg-Andersen, Christian; Barzik, Melanie; Bird, Jonathan E.; Zhao, Bo; Balakrishnan, Lata; Friedman, Thomas B.; Perrin, Benjamin J.

doi:10.1016/j.cub.2020.12.006

Cited by 25 publications

(57 citation statements)

References 69 publications

(81 reference statements)

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“…Disruption of transduction also affects distribution of key bundle proteins (Krey et al, 2020;McGrath et al, 2021). Finally, Ca 2+ entry through transduction channels regulates the function of many stereocilia proteins (Fettiplace, 2017).…”

Section: Introductionmentioning

confidence: 99%

Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

Halford

Bateschell

Barr-Gillespie

2022

MBoC

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BAIAP2L2, a membrane-binding protein required for maintenance of mechanotransduction in hair cells, is selectively retained at tips of transducing stereocilia. BAIAP2L2 trafficked to stereocilia tips in the absence of EPS8, but EPS8 increased the efficiency of localization. A tripartite complex of BAIAP2L2, EPS8, and MYO15A formed efficiently in vitro, and these three proteins robustly targeted to filopodia tips when co-expressed in cultured cells. Mice lacking functional transduction channels no longer concentrated BAIAP2L2 at row 2 stereocilia tips, a result that was phenocopied by blocking channels with tubocurarine in cochlear explants. Transduction channels permit Ca2+ entry into stereocilia, and we found that membrane localization of BAIAP2L2 was enhanced in the presence of Ca2+. Finally, reduction of intracellular Ca2+ in hair cells using BAPTA-AM led to a loss of BAIAP2L2 at stereocilia tips. Taken together, our results show that a MYO15A-EPS8 complex transports BAIAP2L2 to stereocilia tips, and Ca2+ entry through open channels at row 2 tips retains BAIAP2L2 there.

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Section: Introductionmentioning

confidence: 99%

Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

Halford

Bateschell

Barr-Gillespie

2022

MBoC

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“…The stereocilia length was then calculated using the equations mentioned in the above report. Stereocilia width quantification were performed as previously described (McGrath et al, 2021). Briefly, stereocilia with approximately perpendicular orientation in SEM images were used for calculations.…”

Section: Scanning Electron Microscopy Andmentioning

confidence: 99%

“…The subcellular localization of purified protein in permeabilized hair cells was examined as described before (McGrath et al, 2021). All steps were performed at room temperature unless otherwise indicated.…”

Section: Protein Purification and Localization In Permeabilized Hair Cellsmentioning

confidence: 99%

“…We made use of two point mutations that mimic CDC42 in different state, namely inactive CDC42 mutant (T17N) and constitutively active CDC42 mutant (Q61L) (Kozma et al, 1995). We first examined whether exogenous CDC42 mutants could target to the stereocilia following permeabilization of unfixed cochlear sensory epithelia with 0.05% saponin (McGrath et al, 2021). The results show that wild-type CDC42 and two CDC42 mutants all localize in the stereocilia of P0.5 mice, with enrichment at the stereociliary tips (Figure 4A).…”

Section: Constitutively Active Mutant Cdc42 Leads To Enhanced Stereocilia Deficitsmentioning

confidence: 99%

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The Rho GTPase Cell Division Cycle 42 Regulates Stereocilia Development in Cochlear Hair Cells

Zhou

Sun

et al. 2021

Front. Cell Dev. Biol.

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Stereocilia are actin-based cell protrusions on the apical surface of inner ear hair cells, playing a pivotal role in hearing and balancing sensation. The development and maintenance of stereocilia is tightly regulated and deficits in this process usually lead to hearing or balancing disorders. The Rho GTPase cell division cycle 42 (CDC42) is a key regulator of the actin cytoskeleton. It has been reported to localize in the hair cell stereocilia and play important roles in stereocilia maintenance. In the present work, we utilized hair cell-specific Cdc42 knockout mice and CDC42 inhibitor ML141 to explore the role of CDC42 in stereocilia development. Our data show that stereocilia height and width as well as stereocilia resorption are affected in Cdc42-deficient cochlear hair cells when examined at postnatal day 8 (P8). Moreover, ML141 treatment leads to planar cell polarity (PCP) deficits in neonatal hair cells. We also show that overexpression of a constitutively active mutant CDC42 in cochlear hair cells leads to enhanced stereocilia developmental deficits. In conclusion, the present data suggest that CDC42 plays a pivotal role in regulating hair cell stereocilia development.

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“…This sophisticated architecture ensures the optimal gating of the MET channels located at the lower end of the tip-link, when the stereocilia are deflected toward the taller stereocilia 2 . Remarkably, the MET molecular apparatus co-exists at stereocilia tips with the actin regulatory protein complex that sustains the stereocilia length differential, and there is increasing evidence of coordination between these two molecular machineries [3][4][5] .…”

Section: Introductionmentioning

confidence: 99%

Selective binding and transport of protocadherin 15 isoforms by stereocilia unconventional myosins in a heterologous expression system

Ballesteros¹,

Yadav

Cui

et al. 2022

Preprint

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During hair cell development, the mechanoelectrical transduction (MET) apparatus is assembled at the stereocilia tips, where it coexists with the stereocilia actin regulatory machinery. While the myosin-based tipward transport of actin regulatory proteins is well studied, isoform complexity and built-in redundancies in the MET apparatus have limited our understanding of how MET components are transported. We used a heterologous expression system to elucidate the myosin selective transport of isoforms of protocadherin 15 (PCDH15), the protein that mechanically gates the MET apparatus. We show that MYO7A selectively transports the CD3 isoform while MYO3A and MYO3B transports the CD2 isoform. Furthermore, MYO15A showed an insignificant role in the transport of PCDH15, and none of the myosins tested transport PCDH15-CD1. Our data suggest an important role for MYO3A, MYO3B, and MYO7A in the MET apparatus formation and highlight the intricate nature of MET and actin regulation during development and functional maturation of the stereocilia bundle.

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Actin at stereocilia tips is regulated by mechanotransduction and ADF/cofilin

Cited by 25 publications

References 69 publications

Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

The Rho GTPase Cell Division Cycle 42 Regulates Stereocilia Development in Cochlear Hair Cells

Selective binding and transport of protocadherin 15 isoforms by stereocilia unconventional myosins in a heterologous expression system

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Actin at stereocilia tips is regulated by mechanotransduction and ADF/cofilin

Cited by 25 publications

References 69 publications

Ca2+ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

Ca2+ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

The Rho GTPase Cell Division Cycle 42 Regulates Stereocilia Development in Cochlear Hair Cells

Selective binding and transport of protocadherin 15 isoforms by stereocilia unconventional myosins in a heterologous expression system

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Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips

Ca²⁺ entry through mechanotransduction channels localizes BAIAP2L2 to stereocilia tips