Open AccessFlow Cytometric Analysis of the Effects of High Protein Diet On Isolated Rat Liver Mitochondria
Author(s) -
O'Connor J. E.,
Kimler B. F.,
Vargas J. L.
Publication year - 1988
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/j.1365-2184.1988.tb00791.x
Subject(s) - rhodamine 123 , mitochondrion , organelle , biology , fluorescence , rhodamine , differential centrifugation , centrifugation , flow cytometry , biochemistry , microbiology and biotechnology , physics , quantum mechanics , multiple drug resistance , antibiotics
. We have investigated changes that occur in mitochondria obtained from the livers of rats that had been maintained on a high protein diet (80% casein instead of 20%) for 6 months. Liver homogenates were separated by centrifugation into a mitochondrial fraction, a nuclear fraction and the supernatant fluid of the nuclear fraction (nuclear wash). Rhodamine‐123 was used to selectively stain mitochondria depending upon their membrane. potential. the stained organelles were processed through a flow cytometer where the fluorescent stains were excited by the 488 nm wavelength of a laser and the resultant fluorescence signals analysed. After 6 months on a high protein diet, mitochondria displayed an increase in the fluorescence associated with rhodamine‐123 uptake in both mitochondrial and nuclear wash fractions, while mitochondrial fluorescence in the nuclear fraction showed a heterogeneous distribution. This was interpreted as an increase in membrane potential in most of the liver mitochondria under these nutritional conditions, with a certain degree of heterogeneity. These functional changes may be correlated with morphological alterations previously reported and show the usefulness of flow cytometry for biochemical analysis of isolated mitochondria.