High-resolution immunofluorescence imaging of mouse cochlear hair bundles

被引：3

作者：

Miller, Katharine K. ^{[1
]}

Wang, Pei ^{[1
]}

Grillet, Nicolas ^{[1
]}

机构：

[1] Stanford Univ, Dept Otolaryngol Head & Neck Surg, Stanford, CA 94305 USA

来源：

STAR PROTOCOLS | 2022年 / 3卷 / 02期

关键词：

MECHANOTRANSDUCTION MACHINERY; STEREOCILIA; CHANNEL; CELLS; TRANSDUCTION; COMPONENT; LINKS; TMIE;

D O I：

10.1016/j.xpro.2022.101431

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

High-resolution immunofluorescence imaging of cochlear hair bundles faces many challenges due to the hair bundle's small dimensions, fragile nature, and complex organization. Here, we describe an optimized protocol for hair-bundle protein immunostaining and localization. We detail the steps and solutions for extracting and fixing the mouse inner ear and for dissecting the organ of Corti. We further emphasize the optimal permeabilization, blocking, staining, and mounting conditions as well as the parameters for high-resolution microscopy imaging.For complete details on the use and execution of this protocol, please refer to Trouillet et al. (2021).

引用

页数：22

共 19 条

[1] TIP-LINK INTEGRITY AND MECHANICAL TRANSDUCTION IN VERTEBRATE HAIR-CELLS [J].

ASSAD, JA ;

SHEPHERD, GMG ;

COREY, DP .

NEURON, 1991, 7 (06) :985-994

[2] TMIE Defines Pore and Gating Properties of the Mechanotransduction Channel of Mammalian Cochlear Hair Cells [J].

Cunningham, Christopher L. ;

Qiu, Xufeng ;

Wu, Zizhen ;

Zhao, Bo ;

Peng, Guihong ;

Kim, Ye-Hyun ;

Lauer, Amanda ;

Mueller, Ulrich .

NEURON, 2020, 107 (01) :126-+

[3] The dimensions and structural attachments of tip links in mammalian cochlear hair cells and the effects of exposure to different levels of extracellular calcium [J].

Furness, D. N. ;

Katori, Y. ;

Kumar, B. Nirmal ;

Hackney, C. M. .

NEUROSCIENCE, 2008, 154 (01) :10-21

[4] CIB2 interacts with TMC1 and TMC2 and is essential for mechanotransduction in auditory hair cells [J].

Giese, Arnaud P. J. ;

Tang, Yi-Quan ;

Sinha, Ghanshyam P. ;

Bowl, Michael R. ;

Goldring, Adam C. ;

Parker, Andrew ;

Freeman, Mary J. ;

Brown, Steve D. M. ;

Riazuddin, Saima ;

Fettiplace, Robert ;

Schafer, William R. ;

Frolenkov, Gregory I. ;

Ahmed, Zubair M. .

NATURE COMMUNICATIONS, 2017, 8

[5] COMPLIANCE OF THE HAIR BUNDLE ASSOCIATED WITH GATING OF MECHANOELECTRICAL TRANSDUCTION CHANNELS IN THE BULLFROGS SACCULAR HAIR CELL [J].

HOWARD, J ;

HUDSPETH, AJ .

NEURON, 1988, 1 (03) :189-199

[6] Mechanotransduction in mouse inner ear hair cells requires transmembrane channel-like genes [J].

Kawashima, Yoshiyuki ;

Geleoc, Gwenaelle S. G. ;

Kurima, Kiyoto ;

Labay, Valentina ;

Lelli, Andrea ;

Asai, Yukako ;

Makishima, Tomoko ;

Wu, Doris K. ;

Della Santina, Charles C. ;

Holt, Jeffrey R. ;

Griffith, Andrew J. .

JOURNAL OF CLINICAL INVESTIGATION, 2011, 121 (12) :4796-4809

[7] Characterization of the transmembrane channel-like (TMC) gene family:: functional clues from hearing loss and epidermodysplasia verruciformis [J].

Kurima, K ;

Yang, YD ;

Sorber, K ;

Griffith, AJ .

GENOMICS, 2003, 82 (03) :300-308

[8] A Bundle of Mechanisms: Inner-Ear Hair-Cell Mechanotransduction [J].

Maoileidigh, Daibhid O. ;

Ricci, Anthony J. .

TRENDS IN NEUROSCIENCES, 2019, 42 (03) :221-236

[9] Dimensions of a Living Cochlear Hair Bundle [J].

Miller, Katharine K. ;

Atkinson, Patrick ;

Mendoza, Kyssia Ruth ;

Maoileidigh, Daibhid ;

Grillet, Nicolas .

FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY, 2021, 9

[10] Whole Mount Dissection and Immunofluorescence of the Adult Mouse Cochlea [J].

Montgomery, Scott C. ;

Cox, Brandon C. .

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2016, (107)

← 1 2 →