ArticlePloS one2013
A simple method for purification of vestibular hair cells and non-sensory cells, and application for proteomic analysis.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 21 citations in OpenAlex.
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- Molecular and functional profiling of cell diversity and identity in the lateral superior olive, an auditory brainstem center with ascending and descending projections.Frontiers in cellular neuroscience · 2024Article
- Cell Type-Specific Expression Analysis of the Inner Ear: A Technical Report.The Laryngoscope · 2021Article
- Article
- Downregulated UCHL1 Accelerates Gentamicin-Induced Auditory Cell Death via Autophagy.Molecular neurobiology · 2019Article
- Generation of Vestibular Tissue-Like Organoids From Human Pluripotent Stem Cells Using the Rotary Cell Culture System.Frontiers in cell and developmental biology · 2019Article
- Mass spectrometry quantitation of proteins from small pools of developing auditory and vestibular cells.Scientific data · 2018Article
- Label-free quantitative mass spectrometry analysis of differential protein expression in the developing cochlear sensory epithelium.Proteome science · 2018Article
- Global Analysis of Protein Expression of Inner Ear Hair Cells.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2017Article
- Activity-Dependent Phosphorylation by CaMKIIδ Alters the CaFrontiers in synaptic neuroscience · 2017Article
- Applications for single cell trajectory analysis in inner ear development and regeneration.Cell and tissue research · 2015Review
- Mechanisms of otoconia and otolith development.Developmental dynamics : an official publication of the American Association of Anatomists · 2015Review
- The transcriptome of utricle hair cell regeneration in the avian inner ear.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2014Article
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
Mechanosensitive hair cells and supporting cells comprise the sensory epithelia of the inner ear. The paucity of both cell types has hampered molecular and cell biological studies, which often require large quantities of purified cells. Here, we report a strategy allowing the enrichment of relatively pure populations of vestibular hair cells and non-sensory cells including supporting cells. We utilized specific uptake of fluorescent styryl dyes for labeling of hair cells. Enzymatic isolation and flow cytometry was used to generate pure populations of sensory hair cells and non-sensory cells. We applied mass spectrometry to perform a qualitative high-resolution analysis of the proteomic makeup of both the hair cell and non-sensory cell populations. Our conservative analysis identified more than 600 proteins with a false discovery rate of <3% at the protein level and <1% at the peptide level. Analysis of proteins exclusively detected in either population revealed 64 proteins that were specific to hair cells and 103 proteins that were only detectable in non-sensory cells. Statistical analyses extended these groups by 53 proteins that are strongly upregulated in hair cells versus non-sensory cells and vice versa by 68 proteins. Our results demonstrate that enzymatic dissociation of styryl dye-labeled sensory hair cells and non-sensory cells is a valid method to generate pure enough cell populations for flow cytometry and subsequent molecular analyses.
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