Abstract
Hair cells of the inner ear 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 how the proteome changes over development from postnatal days 0 through 7, we isolated pools of 5000 Pou4f3-Gfp positive or negative cells from the cochlea or utricles; these cell pools were analysed 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. DIA measurements were extremely sensitive; we were able to detect proteins present at less than one part in 100,000 from only 312 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.
Highlights
Background & SummaryHair cells, the sensory cells of the inner ear, are responsible for detection, amplification, and transmission of auditory and vestibular stimuli[1]
The DDA and DIA datasets will be valuable for accurately quantifying proteins in hair cells and non-hair cells over this developmental window
In a mouse cochlea, which mediates auditory transduction, ~3000 hair cells reside in the organ of Corti, a small organ of about a dozen cell types that sits on the vibrating basilar membrane[4,5]
Summary
Background & SummaryHair cells, the sensory cells of the inner ear, are responsible for detection, amplification, and transmission of auditory and vestibular stimuli[1]. The Pou4f3-GFP mouse line has been used for measurement of protein abundance in experiments similar to the transcript analysis described above; mass spectrometry analysis of large pools of hair cells sorted by FACS allowed the identification of >6000 proteins, including >900 expressed in hair cells[13].
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