Proteomics

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Embryonic day 15 chick utricle single cell analysis

ABSTRACT: Single-cell proteomics can reveal the changing protein composition of differentiating cells. We used shotgun mass spectrometry to determine the abundant proteins present in single or small pools of subpicoliter-sized cells from the embryonic day 15 (E15) utricle of the chicken inner ear, when many hair cells are differentiating from progenitor (supporting) cells. The actin monomer binding protein thymosin β4 (TMSB4X) was present in E15 progenitor cells at nearly equimolar levels relative to actin, but dropped to one-tenth that value in hair cells, with little change in total actin. Single-cell RNA-seq analysis of E15 utricle cells showed that TMSB4X transcripts fell in abundance once hair-cell differentiation initiated. These results suggest that most actin is sequestered in progenitor cells, but upon differentiation to hair cells, actin is released, permitting assembly of the sensory hair bundle.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Gallus Gallus (chicken)

TISSUE(S): Inner Ear Vestibulum, Ear Hair Cell, Inner Ear, Vestibular Hair Cell

SUBMITTER: Peter Barr-Gillespie  

LAB HEAD: Peter Barr-Gillespie

PROVIDER: PXD014256 | Pride | 2019-11-12

REPOSITORIES: Pride

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700526_Single_Hair_Cell_OHSU_20cell_High_090618_1_30um_031218_300bar.raw Raw
700533_Single_Hair_Cell_OHSU_20cell_High_090618_2_30um_031218_300bar.raw Raw
700539_Single_Hair_Cell_OHSU_20cell_High_090618_3_30um_031218_300bar.raw Raw
700567_Single_Hair_Cell_OHSU_20cell_High_090618_4_30um_031218_300bar.raw Raw
700576_Single_Hair_Cell_OHSU_5cell_High_090618_1_30um_031218_300bar.raw Raw
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Publications

Single-cell proteomics reveals changes in expression during hair-cell development.

Zhu Ying Y   Scheibinger Mirko M   Ellwanger Daniel Christian DC   Krey Jocelyn F JF   Choi Dongseok D   Kelly Ryan T RT   Heller Stefan S   Barr-Gillespie Peter G PG  

eLife 20191104

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 developme  ...[more]

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