Renal proximal tubule cells have a tremendous capacity to recover from cell injury such as ischemia. We have analyzed the changes to myosin 1 in a reversible model of ATP depletion using substrate depleted media and antimycin A in confluent LLCPKi cells. Previous studies by this laboratory and others have documented the rapid disorganization of the actin cytoskeleton following an ischémie event and importantly, the cells ability to recover following removal of the cells or tissue from the adverse conditions. Our analysis has utilized both indirect immunofluorescent microscopy and biochemical detergent fractionation to assess the changes occurring to myosin I during this cell injury event. We find that myosin I is present in a widespread punctate distribution in normal cells including the microvilli. Following ATP depletion there is a shift detected by both immunofluorescence and biochemical extraction techniques to increased association with the F-actin cytoskeleton. F-actin content increases with accumulation of cytosolic aggregates, and myosin I is concentrated in these aggregates. We are presently quantifying the biochemical changes of this motor and analyzing the recovery process. In addition, we are interested in addressing potential posttranslational events which may contribute to the distribution changes we are observing.
|Original language||English (US)|
|Journal||Journal of Investigative Medicine|
|State||Published - Jan 1 1996|
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)