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/sup 3/H-ouabain binding and sodium-pump activity measured in myocytes isolated from guinea-pig heart

Thesis/Dissertation ·
OSTI ID:6401140
Because of the toxicity of millimolar ouabain, non-specific /sup 3/H-ouabain binding was assessed by monitoring the dissociation of the bound drug. Analysis of specific /sup 3/H-ouabain binding to myocytes yielded non-linear Scatchard plots. Nonlinearity appears to result from reduced /sup 3/H-ouabain binding due to low intracellular Na/sup 2/ concentration. Addition of 2 ..mu..M monensin, A Na/sup +/ ionophore, significantly increased /sup 3/H-ouabain binding. Incubation in Ca/sup 2 +/-free solution (0.25 mM EGTA) stimulated /sup 3/H-ouabain binding to a greater degree than monensin and caused Scatchard plots to have two distinct linear components. Monensin had no significant effects when /sup 3/H-ouabain binding occurred in Ca/sup 2 +/-free solution. Effects of Ca/sup 2 +/-free incubation to increase /sup 3/H-ouabain binding suggest that Ca/sup 2 +/ has a direct effect on /sup 3/H-ouabain binding. Alternatively, Ca/sup 2 +/-free incubation may increase Na/sup +/ permeability of the sarcolemma. Isoproterenol, phenylephrine, TPA (phorbol 12-myristate 13-acetate), La/sup 3 +/, and the Ca/sup 2 +/-ionophore A23187 failed to cause significant changes in /sup 3/H-ouabain binding when myocytes were incubated in a solution containing 0.5 or 2.5 ..mu..M /sup 3/H-ouabain, 0.1 mM Ca/sup 2 +/ and 1 mM K/sup +/.
Research Organization:
Michigan State Univ., East Lansing (USA)
OSTI ID:
6401140
Country of Publication:
United States
Language:
English

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Related Subjects

550201 -- Biochemistry-- Tracer Techniques
560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALKALI METAL COMPOUNDS
ALKALINE EARTH METAL COMPOUNDS
ANIMAL CELLS
ANIMALS
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BIOLOGICAL FUNCTIONS
BODY
CALCIUM COMPOUNDS
CARBOHYDRATES
CARCINOGENS
CARDIAC GLYCOSIDES
CARDIOTONICS
CARDIOVASCULAR AGENTS
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL MEMBRANES
DRUGS
ESTERS
FUNCTIONS
GLYCOSIDES
GUINEA PIGS
HEART
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE TRANSPORT
MEMBRANES
ORGANIC COMPOUNDS
ORGANS
OUABAIN
PHORBOL ESTERS
REACTION KINETICS
RODENTS
SODIUM COMPOUNDS
STROPHANTHINS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES