Mass spectrometry identification of proteins of isolated auditory and vestibular hair cells using data-independent and data-dependent acquisition

ABSTRACT: Hair cells undergo postnatal development that leads to formation of their sensory organelles, synaptic machinery, and in the case of cochlear outer hair cells, their electromotile mechanism. To examine the proteome changes over development, we isolated pools of 5000 Pou4f3-Gfp positive or negative cells from the cochlea or utricles; these cell pools were analyzed by data-dependent and data-independent acquisition (DDA and DIA) mass spectrometry. DDA data were used to generate spectral libraries, which enabled identification and accurate quantitation of specific proteins using the DIA datasets. We also isolated and pooled individual inner and outer hair cells from adult cochlea and compared their proteomes to those of developing hair cells. The DDA and DIA datasets will be valuable for accurately quantifying proteins in hair cells and non-hair cells over this developmental window.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Supporting Cell Of Cochlea, Supporting Cell Of Vestibular Epithelium, Ear Hair Cell, Cochlea, Auditory Hair Cell, Utricle, Inner Ear, Vestibular Hair Cell

SUBMITTER: Peter Barr-Gillespie  

LAB HEAD: Peter Barr-Gillespie

PROVIDER: PXD006240 | Pride | 2017-09-18

REPOSITORIES: Pride