Cy3-ATP labeling of unfixed, permeabilized mouse hair cells.

ABSTRACT: ATP-utilizing enzymes play key roles in hair bundles, the mechanically sensitive organelles of sensory hair cells in the inner ear. We used a fluorescent ATP analog, EDA-ATP-Cy3 (Cy3-ATP), to label ATP-binding proteins in two different preparations of unfixed hair-cell stereocilia of the mouse. In the first preparation, we lightly permeabilized dissected cochleas, then labeled them with Cy3-ATP. Hair cells and their stereocilia remained intact, and stereocilia tips in rows 1 and 2 were labeled particularly strongly with Cy3-ATP. In many cases, vanadate (V_i) traps nucleotides at the active site of myosin isoforms and presents nucleotide dissociation. Co-application with V_i enhanced the tip labeling, which is consistent with myosin isoforms being responsible. By contras

SUBMITTER: Pacentine IV 

PROVIDER: S-EPMC8668996 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature