Isotachophoretic analyses of metabolites of cardiac and skeletal muscles in four species.

Abstract

We studied ATP and concentrations of other metabolites in cardiac and skeletal muscles from different species (frog, hamster, guinea-pig, and dog), using analytical capillary isotachophoresis. The method had several advantages for quantitative analysis of tissue metabolites : short separation time, high sensitivity, high resolution, and good reproducibility. It was also possible to detect a number of compounds simultaneously, including ATP, ADP, AMP, cyclic AMP, creatine phosphate (CP), inosine monophosphate, NAD, NADH, glucose-6-phosphate, pyruvate, and inorganic phosphate. Generally, the concentrations of high-energy phosphates (ATP and CP) in the cardiac muscle were significantly lower than those in the skeletal muscle, in all species tested, except for hamster where the concentration of CP in the skeletal muscle was comparable to that in the cardiac muscle. In mammals, ATP and CP concentrations were comparable in the atrium and ventricle while the concentration in the frog ventricle was significantly higher than that in the atrium. The data obtained by this isotachophoretic analysis were comparable to the data reported by the use of other conventional analytical methods. The significance and reliability of isotachophoresis in the determination of the various metabolites in the cardiac muscle were discussed. Key Words: isotachophoresis, metabolites, cardiac and skeletal muscles.Analytical isotachophoresis is a pertinent tool for the analysis of various chemical compounds and has several advantages, e.g., short separation time, high sensitivity, high resolution, and good reproducibility. With this method, it is possible to detect a large number of metabolites during one single run. This approach has been used to analyze proteins (

Fiber Composition and Oxidative Capacity of Hamster Skeletal Muscle

“…The hamster has been utilized to investigate pathological dysfunction stemming from diabetes mellitus (Sims and Landau 1967), hypoglycemia (Sodoyez et al 1969), muscular dystrophy (Lochner et al 1971), cardiomyopathy (Schwartz et al 1972), hypothermia (Resch and Musacchia 1976), emphysema (Karlinsky and Snider 1978), skeletal muscle transplantation (Faulkner et al 1983), age (Nichols and Borer 1987), chronic wasting infection (Drew et al 1988), skeletal muscle ischemia/reperfusion (Messina 1990), inactivity (Zhan and Sieck 1992), and gene therapy (Xiao et al 2000). Moreover, the hamster has been used to explore the physiological function of the myocardium (Aomine et al 1982), vasculature (Burns and Palade 1968), and skeletal muscle (Lossnitzer and Kelly 1968). In other animal species, investigations integrating the cardiorespiratory and muscular systems have demonstrated the importance of understanding the fiber composition of skeletal muscle.…”

Surface layer ATP-related contraction in isolated,superfused canine ventricular papillary muscle: An isotachophoretic analysis.

“…We examined the relationships among the contractile tension and the content and concentration of various metabolites, using canine papillary muscles of different sizes exposed to either normoxic or hypoxic conditions. For determination of metabolite concentrations, we used capillary isotachophoresis which enabled simultaneous measurement of many metabolites (including adenine nucleotides) contained in a single muscle preparation (AOMINE et al, 1982). Among these metabolites, special attention was directed to the relation between ATP levels and contractile tension, in attempts to demonstrate the existence or absence of the anoxic core.…”