Single-cell proteomics reveals changes in expression during hair-cell development
Abstract
Hearing and balance rely on small sensory hair cells that reside in the inner ear. To explore dynamic changes in the abundant proteins present in differentiating hair cells, we used nanoliter-scale shotgun mass spectrometry of single cells, each ~1 picoliter, from utricles of embryonic day 15 chickens. We identified unique constellations of proteins or protein groups from presumptive hair cells and from progenitor cells. The single-cell proteomes enabled the de novo reconstruction of a developmental trajectory using protein expression levels, revealing proteins that greatly increased in expression during differentiation of hair cells (e.g., OCM, CRABP1, GPX2, AK1, GSTO1) and those that decreased during differentiation (e.g., TMSB4X, AGR3). Complementary single-cell transcriptome profiling showed corresponding changes in mRNA during maturation of hair cells. Single-cell proteomics data thus can be mined to reveal features of cellular development that may be missed with transcriptomics.
- Authors:
-
- Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, United States
- Department of Otolaryngology Head and Neck Surgery, Stanford University, Stanford, United States
- Department of Otolaryngology Head and Neck Surgery, Stanford University, Stanford, United States, Genome Analysis Unit, Amgen Research, Amgen Inc, South San Francisco, United States
- Oregon Hearing Research Center, Oregon Health & Science University, Portland, United States, Vollum Institute, Oregon Health & Science University, Portland, United States
- OHSU-PSU School of Public Health, Oregon Health & Science University, Portland, United States, Graduate School of Dentistry, Kyung Hee University, Seoul, Republic of Korea
- Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, United States, Department of Chemistry and Biochemistry, Brigham Young University, Provo, United States
- Publication Date:
- Research Org.:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1592800
- Alternate Identifier(s):
- OSTI ID: 1575404; OSTI ID: 1592801
- Report Number(s):
- PNNL-SA-144744
Journal ID: ISSN 2050-084X; e50777
- Grant/Contract Number:
- Laboratory Directed Research and Development Program (Earth & Biological Sciences Directorate Mission Seed); AC05-76RL01830
- Resource Type:
- Journal Article: Published Article
- Journal Name:
- eLife
- Additional Journal Information:
- Journal Name: eLife Journal Volume: 8; Journal ID: ISSN 2050-084X
- Publisher:
- eLife Sciences Publications, Ltd.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Zhu, Ying, Scheibinger, Mirko, Ellwanger, Daniel Christian, Krey, Jocelyn F., Choi, Dongseok, Kelly, Ryan T., Heller, Stefan, and Barr-Gillespie, Peter G. Single-cell proteomics reveals changes in expression during hair-cell development. United States: N. p., 2019.
Web. doi:10.7554/eLife.50777.
Zhu, Ying, Scheibinger, Mirko, Ellwanger, Daniel Christian, Krey, Jocelyn F., Choi, Dongseok, Kelly, Ryan T., Heller, Stefan, & Barr-Gillespie, Peter G. Single-cell proteomics reveals changes in expression during hair-cell development. United States. https://doi.org/10.7554/eLife.50777
Zhu, Ying, Scheibinger, Mirko, Ellwanger, Daniel Christian, Krey, Jocelyn F., Choi, Dongseok, Kelly, Ryan T., Heller, Stefan, and Barr-Gillespie, Peter G. Mon .
"Single-cell proteomics reveals changes in expression during hair-cell development". United States. https://doi.org/10.7554/eLife.50777.
@article{osti_1592800,
title = {Single-cell proteomics reveals changes in expression during hair-cell development},
author = {Zhu, Ying and Scheibinger, Mirko and Ellwanger, Daniel Christian and Krey, Jocelyn F. and Choi, Dongseok and Kelly, Ryan T. and Heller, Stefan and Barr-Gillespie, Peter G.},
abstractNote = {Hearing and balance rely on small sensory hair cells that reside in the inner ear. To explore dynamic changes in the abundant proteins present in differentiating hair cells, we used nanoliter-scale shotgun mass spectrometry of single cells, each ~1 picoliter, from utricles of embryonic day 15 chickens. We identified unique constellations of proteins or protein groups from presumptive hair cells and from progenitor cells. The single-cell proteomes enabled the de novo reconstruction of a developmental trajectory using protein expression levels, revealing proteins that greatly increased in expression during differentiation of hair cells (e.g., OCM, CRABP1, GPX2, AK1, GSTO1) and those that decreased during differentiation (e.g., TMSB4X, AGR3). Complementary single-cell transcriptome profiling showed corresponding changes in mRNA during maturation of hair cells. Single-cell proteomics data thus can be mined to reveal features of cellular development that may be missed with transcriptomics.},
doi = {10.7554/eLife.50777},
url = {https://www.osti.gov/biblio/1592800},
journal = {eLife},
issn = {2050-084X},
number = ,
volume = 8,
place = {United States},
year = {2019},
month = {11}
}
Web of Science
Works referenced in this record:
Small actin-binding proteins: the β-thymosin family
journal, February 1993
- Nachmias, Vivianne T.
- Current Opinion in Cell Biology, Vol. 5, Issue 1
Plastin 1 widens stereocilia by transforming actin filament packing from hexagonal to liquid
journal, November 2016
- Krey, Jocelyn F.; Krystofiak, Evan S.; Dumont, Rachel A.
- The Journal of Cell Biology, Vol. 215, Issue 4
Actin monomer binding proteins
journal, January 1995
- Sun, Hui-Qiao; Kwiatkowska, Katarzyna; Yin, Helen L.
- Current Opinion in Cell Biology, Vol. 7, Issue 1
Current best practices in single‐cell RNA‐seq analysis: a tutorial
journal, June 2019
- Luecken, Malte D.; Theis, Fabian J.
- Molecular Systems Biology, Vol. 15, Issue 6
SCnorm: robust normalization of single-cell RNA-seq data
journal, April 2017
- Bacher, Rhonda; Chu, Li-Fang; Leng, Ning
- Nature Methods, Vol. 14, Issue 6
Global quantification of mammalian gene expression control
journal, May 2011
- Schwanhäusser, Björn; Busse, Dorothea; Li, Na
- Nature, Vol. 473, Issue 7347
Actin Filaments, Stereocilia, and Hair Cells: How Cells Count and Measure
journal, November 1992
- Tilney, L. G.; Tilney, M. S.; DeRosier, D. J.
- Annual Review of Cell Biology, Vol. 8, Issue 1
The PRIDE database and related tools and resources in 2019: improving support for quantification data
journal, November 2018
- Perez-Riverol, Yasset; Csordas, Attila; Bai, Jingwen
- Nucleic Acids Research, Vol. 47, Issue D1
Transcriptional Dynamics of Hair-Bundle Morphogenesis Revealed with CellTrails
journal, June 2018
- Ellwanger, Daniel C.; Scheibinger, Mirko; Dumont, Rachel A.
- Cell Reports, Vol. 23, Issue 10
β-Actin and γ-Actin Are Each Dispensable for Auditory Hair Cell Development But Required for Stereocilia Maintenance
journal, October 2010
- Perrin, Benjamin J.; Sonnemann, Kevin J.; Ervasti, James M.
- PLoS Genetics, Vol. 6, Issue 10
Hair Bundles Are Specialized for ATP Delivery via Creatine Kinase
journal, February 2007
- Shin, Jung-Bum; Streijger, Femke; Beynon, Andy
- Neuron, Vol. 53, Issue 3
A multiregional proteomic survey of the postnatal human brain
journal, November 2017
- Carlyle, Becky C.; Kitchen, Robert R.; Kanyo, Jean E.
- Nature Neuroscience, Vol. 20, Issue 12
Proteomic Analysis of Single Mammalian Cells Enabled by Microfluidic Nanodroplet Sample Preparation and Ultrasensitive NanoLC-MS
journal, June 2018
- Zhu, Ying; Clair, Geremy; Chrisler, William B.
- Angewandte Chemie International Edition, Vol. 57, Issue 38
Roles of the Espin Actin-Bundling Proteins in the Morphogenesis and Stabilization of Hair Cell Stereocilia Revealed in CBA/CaJ Congenic Jerker Mice
journal, March 2011
- Sekerková, Gabriella; Richter, Claus-Peter; Bartles, James R.
- PLoS Genetics, Vol. 7, Issue 3
Nanog suppresses cell migration by downregulating thymosin β4 and Rnd3
journal, January 2013
- Zhou, Yizhuo; Li, Shanshan; Huang, Qiongping
- Journal of Molecular Cell Biology, Vol. 5, Issue 4
High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform
journal, September 2019
- Dou, Maowei; Clair, Geremy; Tsai, Chia-Feng
- Analytical Chemistry, Vol. 91, Issue 20
PDZD7-MYO7A complex identified in enriched stereocilia membranes
journal, August 2016
- Morgan, Clive P.; Krey, Jocelyn F.; Grati, M'hamed
- eLife, Vol. 5
A test metric for assessing single-cell RNA-seq batch correction
journal, December 2018
- Büttner, Maren; Miao, Zhichao; Wolf, F. Alexander
- Nature Methods, Vol. 16, Issue 1
Identification of the Hair Cell Soma-1 Antigen, HCS-1, as Otoferlin
journal, August 2010
- Goodyear, Richard J.; Legan, P. Kevin; Christiansen, Jeffrey R.
- Journal of the Association for Research in Otolaryngology, Vol. 11, Issue 4
Correlation of Actin Crosslinker and Capper Expression Levels with Stereocilia Growth Phases
journal, December 2013
- Avenarius, Matthew R.; Saylor, Katherine W.; Lundeberg, Megan R.
- Molecular & Cellular Proteomics, Vol. 13, Issue 2
MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification
journal, November 2008
- Cox, Jürgen; Mann, Matthias
- Nature Biotechnology, Vol. 26, Issue 12
Full-length RNA-seq from single cells using Smart-seq2
journal, January 2014
- Picelli, Simone; Faridani, Omid R.; Björklund, Åsa K.
- Nature Protocols, Vol. 9, Issue 1
Intracellular water-specific MR of microbead-adherent cells: the HeLa cell intracellular water exchange lifetime
journal, January 2008
- Zhao, L.; Kroenke, C. D.; Song, J.
- NMR in Biomedicine, Vol. 21, Issue 2
SCoPE-MS: mass spectrometry of single mammalian cells quantifies proteome heterogeneity during cell differentiation
journal, October 2018
- Budnik, Bogdan; Levy, Ezra; Harmange, Guillaume
- Genome Biology, Vol. 19, Issue 1
The control of actin nucleotide exchange by thymosin beta 4 and profilin. A potential regulatory mechanism for actin polymerization in cells.
journal, September 1992
- Goldschmidt-Clermont, P. J.; Furman, M. I.; Wachsstock, D.
- Molecular Biology of the Cell, Vol. 3, Issue 9
Diffusion pseudotime robustly reconstructs lineage branching
journal, August 2016
- Haghverdi, Laleh; Büttner, Maren; Wolf, F. Alexander
- Nature Methods, Vol. 13, Issue 10
Comparative Proteomic Analysis of Eleven Common Cell Lines Reveals Ubiquitous but Varying Expression of Most Proteins
journal, January 2012
- Geiger, Tamar; Wehner, Anja; Schaab, Christoph
- Molecular & Cellular Proteomics, Vol. 11, Issue 3
A Simple Method for Purification of Vestibular Hair Cells and Non-Sensory Cells, and Application for Proteomic Analysis
journal, June 2013
- Herget, Meike; Scheibinger, Mirko; Guo, Zhaohua
- PLoS ONE, Vol. 8, Issue 6
lmerTest Package: Tests in Linear Mixed Effects Models
journal, January 2017
- Kuznetsova, Alexandra; Brockhoff, Per B.; Christensen, Rune H. B.
- Journal of Statistical Software, Vol. 82, Issue 13
Integral membrane proteins: bottom-up, top-down and structural proteomics
journal, May 2017
- Kar, Upendra K.; Simonian, Margaret; Whitelegge, Julian P.
- Expert Review of Proteomics, Vol. 14, Issue 8
Essential nucleotide- and protein-dependent functions of Actb /β-actin
journal, July 2018
- Patrinostro, Xiaobai; Roy, Pallabi; Lindsay, Angus
- Proceedings of the National Academy of Sciences, Vol. 115, Issue 31
New mass spectrometry technologies contributing towards comprehensive and high throughput omics analyses of single cells
journal, January 2019
- Couvillion, Sneha P.; Zhu, Ying; Nagy, Gabe
- The Analyst, Vol. 144, Issue 3
Parvalbumin 3 is an Abundant Ca2+ Buffer in Hair Cells
journal, December 2002
- Heller, Stefan; Bell, Andrea M.; Denis, Charlotte S.
- JARO - Journal of the Association for Research in Otolaryngology, Vol. 3, Issue 4
Andromeda: A Peptide Search Engine Integrated into the MaxQuant Environment
journal, April 2011
- Cox, Jürgen; Neuhauser, Nadin; Michalski, Annette
- Journal of Proteome Research, Vol. 10, Issue 4
Miniaturization Technologies for Efficient Single-Cell Library Preparation for Next-Generation Sequencing
journal, July 2016
- Mora-Castilla, Sergio; To, Cuong; Vaezeslami, Soheila
- Journal of Laboratory Automation, Vol. 21, Issue 4
Adjusting batch effects in microarray expression data using empirical Bayes methods
journal, April 2006
- Johnson, W. Evan; Li, Cheng; Rabinovic, Ariel
- Biostatistics, Vol. 8, Issue 1
Easy to Apply Polyoxazoline-Based Coating for Precise and Long-Term Control of Neural Patterns
journal, May 2017
- Weydert, Serge; Zürcher, Stefan; Tanner, Stefanie
- Langmuir, Vol. 33, Issue 35
Aminoglycoside Damage and Hair Cell Regeneration in the Chicken Utricle
journal, November 2017
- Scheibinger, Mirko; Ellwanger, Daniel C.; Corrales, C. Eduardo
- Journal of the Association for Research in Otolaryngology, Vol. 19, Issue 1
limma powers differential expression analyses for RNA-sequencing and microarray studies
journal, January 2015
- Ritchie, Matthew E.; Phipson, Belinda; Wu, Di
- Nucleic Acids Research, Vol. 43, Issue 7
On the Dependency of Cellular Protein Levels on mRNA Abundance
journal, April 2016
- Liu, Yansheng; Beyer, Andreas; Aebersold, Ruedi
- Cell, Vol. 165, Issue 3
Accurate Label-Free Protein Quantitation with High- and Low-Resolution Mass Spectrometers
journal, December 2013
- Krey, Jocelyn F.; Wilmarth, Phillip A.; Shin, Jung-Bum
- Journal of Proteome Research, Vol. 13, Issue 2
Proteome Profiling of 1 to 5 Spiked Circulating Tumor Cells Isolated from Whole Blood Using Immunodensity Enrichment, Laser Capture Microdissection, Nanodroplet Sample Processing, and Ultrasensitive nanoLC–MS
journal, September 2018
- Zhu, Ying; Podolak, Jennifer; Zhao, Rui
- Analytical Chemistry, Vol. 90, Issue 20
Thymosin β4 suppresses CCl 4 -induced murine hepatic fibrosis by down-regulating transforming growth factor β receptor-II
journal, August 2018
- Li, Hanchao; Li, Qian; Zhang, Xueting
- The Journal of Gene Medicine, Vol. 20, Issue 9
Nanodroplet processing platform for deep and quantitative proteome profiling of 10–100 mammalian cells
journal, February 2018
- Zhu, Ying; Piehowski, Paul D.; Zhao, Rui
- Nature Communications, Vol. 9, Issue 1
Ongoing production of sensory cells in the vestibular epithelium of the chick
journal, January 1992
- Roberson, David F.; Weisleder, Pedro; Bohrer, Pamela S.
- Hearing Research, Vol. 57, Issue 2
-Actin is required for cytoskeletal maintenance but not development
journal, June 2009
- Belyantseva, I. A.; Perrin, B. J.; Sonnemann, K. J.
- Proceedings of the National Academy of Sciences, Vol. 106, Issue 24
Interaction of thymosin .beta.4 with muscle and platelet actin: implications for actin sequestration in resting platelets
journal, July 1992
- Weber, A.; Nachmias, V. T.; Pennise, C. R.
- Biochemistry, Vol. 31, Issue 27
Hair-bundle proteomes of avian and mammalian inner-ear utricles
journal, December 2015
- Wilmarth, Phillip A.; Krey, Jocelyn F.; Shin, Jung-Bum
- Scientific Data, Vol. 2, Issue 1
The R109H Variant of Fascin-2, a Developmentally Regulated Actin Crosslinker in Hair-Cell Stereocilia, Underlies Early-Onset Hearing Loss of DBA/2J Mice
journal, July 2010
- Shin, J. -B.; Longo-Guess, C. M.; Gagnon, L. H.
- Journal of Neuroscience, Vol. 30, Issue 29
Monoclonal antibodies against myofibrillar components of rat skeletal muscle decorate the intermediate filaments of cultured cells.
journal, April 1981
- Lin, J. J.
- Proceedings of the National Academy of Sciences, Vol. 78, Issue 4
A Unique Microglia Type Associated with Restricting Development of Alzheimer’s Disease
journal, June 2017
- Keren-Shaul, Hadas; Spinrad, Amit; Weiner, Assaf
- Cell, Vol. 169, Issue 7
The MaxQuant computational platform for mass spectrometry-based shotgun proteomics
journal, October 2016
- Tyanova, Stefka; Temu, Tikira; Cox, Juergen
- Nature Protocols, Vol. 11, Issue 12
Molecular architecture of the chick vestibular hair bundle
journal, January 2013
- Shin, Jung-Bum; Krey, Jocelyn F.; Hassan, Ahmed
- Nature Neuroscience, Vol. 16, Issue 3
‘Living cantilever arrays’ for characterization of mass of single live cells in fluids
journal, January 2008
- Park, Kidong; Jang, Jaesung; Irimia, Daniel
- Lab on a Chip, Vol. 8, Issue 7
Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing
journal, January 1995
- Benjamini, Yoav; Hochberg, Yosef
- Journal of the Royal Statistical Society: Series B (Methodological), Vol. 57, Issue 1
Posttranslational modification of class III beta-tubulin.
journal, September 1990
- Lee, M. K.; Rebhun, L. I.; Frankfurter, A.
- Proceedings of the National Academy of Sciences, Vol. 87, Issue 18
Subnanogram proteomics: Impact of LC column selection, MS instrumentation and data analysis strategy on proteome coverage for trace samples
journal, April 2018
- Zhu, Ying; Zhao, Rui; Piehowski, Paul D.
- International Journal of Mass Spectrometry, Vol. 427
Loss-less Nano-fractionator for High Sensitivity, High Coverage Proteomics
journal, January 2017
- Kulak, Nils A.; Geyer, Philipp E.; Mann, Matthias
- Molecular & Cellular Proteomics, Vol. 16, Issue 4
The actin filament content of hair cells of the bird cochlea is nearly constant even though the length, width, and number of stereocilia vary depending on the hair cell location
journal, December 1988
- Tilney, L. G.
- The Journal of Cell Biology, Vol. 107, Issue 6