Thyroid hormone effect on human mitochondria measured by flow cytometry. 2009

Jan Kvetny, and Tobias Bomholt, and Palle Pedersen, and Line Wilms, and Stine Anthonsen, and Jacob Larsen
The Mitochondrial Research Unit, Department of Internal Medicine, Department of Clinical Biochemistry Department of Clinical Pathology, Naestved Sygehus, Naestved, Denmark. jkve@regionsjaelland.dk

BACKGROUND Mitochondrial function may be impaired in a number of diseases including metabolic syndrome, cardiovascular disease and endocrine disorders. Therefore it is important to be able to measure mitochondrial function in human cells. OBJECTIVE The aim of the present study was to evaluate a method to measure mitochondrial function in human derived cells, which also would reflect regulation by thyroid hormones. METHODS The MDA-MB-231 cell line (a human breast cancer cell line) was incubated with bioactive iodothyronines (T(4), 3'-3, 5-T(3), 3, 5-T(2)) 50 nmol/l for 3 h. Mitochondrial membrane potentials (MMP) were measured by a flow cytometer after staining with Tetramethylrhodamine methyl ester (TMRM). Also, the effect of TRIAC (a stimulator of thyroid hormone nuclear receptors) and L-Carnitine (an inhibitor of thyroid hormone passage into the nucleus) was examined. RESULTS It was possible to measure mitochondrial membrane potential (MMP) in human derived cells and to examine thyroid hormone effects using flow cytometry. Bioactive iodothyronines increased mitochondrial membrane potential. TRIAC had no effect and L-Carnitine only inhibited T(4) stimulation of membrane potential. CONCLUSIONS Flow cytometry may be a valuable method for examining and testing mitochondrial function in human cells. Our findings demonstrate increase of mitochondrial membrane potential and an extra nuclear short time effect of 3, 5-T(2) on mitochondrial activity.

UI MeSH Term Description Entries
D008928 Mitochondria Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed) Mitochondrial Contraction,Mitochondrion,Contraction, Mitochondrial,Contractions, Mitochondrial,Mitochondrial Contractions
D004952 Esters Compounds derived from organic or inorganic acids in which at least one hydroxyl group is replaced by an –O-alkyl or another organic group. They can be represented by the structure formula RCOOR’ and are usually formed by the reaction between an acid and an alcohol with elimination of water. Ester
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005453 Fluorescence The property of emitting radiation while being irradiated. The radiation emitted is usually of longer wavelength than that incident or absorbed, e.g., a substance can be irradiated with invisible radiation and emit visible light. X-ray fluorescence is used in diagnosis.
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013963 Thyroid Hormones Natural hormones secreted by the THYROID GLAND, such as THYROXINE, and their synthetic analogs. Thyroid Hormone,Hormone, Thyroid,Hormones, Thyroid
D013970 Thyronines A group of metabolites derived from THYROXINE and TRIIODOTHYRONINE via the peripheral enzymatic removal of iodines from the thyroxine nucleus. Thyronine is the thyroxine nucleus devoid of its four iodine atoms. Thyronine
D045744 Cell Line, Tumor A cell line derived from cultured tumor cells. Tumor Cell Line,Cell Lines, Tumor,Line, Tumor Cell,Lines, Tumor Cell,Tumor Cell Lines

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