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Title: Phosphorus-31 NMR magnetization transfer measurements of metabolic reaction rates in the rat heart and kidney in vivo

Abstract

This dissertation is concerned with the measurement of the rates of ATP synthesis in the rat kidney and of the creatine kinase catalyzed reaction in the rat heart in situ. Chronically implanted detection coils, employing a balanced matching configuration of capacitors in the tuned circuit, were used to obtain /sup 31/P NMR spectra from heart, kidney, and liver in situ. Gated spectra of heart obtained at systole and diastole and the effects of fructose on kidney and liver were studied. The ability to observe other nuclei using implanted coils is illustrated with /sup 39/K NMR spectra from kidney and muscle. The theoretical considerations of applying magnetization transfer techniques to intact organs are discussed with emphasis on the problems associated with multiple exchange reactions and compartmentation of reactants. Experimental measurements of the ATP synthesis rate (13 ..mu..mol/min/gm tissue) were compared to whole kidney oxygen consumption and Na/sup +/ reabsorption rates to derive ATP/O (0.8 to 1.7) and Na/sup +//ATP (4 to 10) values. The problems associated with ATP synthesis rate measurements in kidney, e.g., the heterogeneity of the inorganic phosphate resonance, are discussed and experiments to overcome these problems proposed.

Authors:
Publication Date:
Research Org.:
Lawrence Berkeley Lab., CA (USA)
OSTI Identifier:
6451478
Report Number(s):
LBL-18376
ON: DE85001195
DOE Contract Number:
AC03-76SF00098
Resource Type:
Technical Report
Resource Relation:
Other Information: Portions are illegible in microfiche products. Thesis
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ATP; BIOSYNTHESIS; HEART; NUCLEAR MAGNETIC RESONANCE; KIDNEYS; PHOSPHOTRANSFERASES; METABOLISM; BIOCHEMICAL REACTION KINETICS; CREATINE; EXPERIMENTAL DATA; IN VIVO; PHOSPHORUS 31; POTASSIUM 39; RATS; ALKALI METAL ISOTOPES; AMINO ACIDS; ANIMALS; BODY; CARBOXYLIC ACIDS; CARDIOVASCULAR SYSTEM; DATA; ENZYMES; INFORMATION; ISOTOPES; KINETICS; LIGHT NUCLEI; MAGNETIC RESONANCE; MAMMALS; NUCLEI; NUCLEOTIDES; NUMERICAL DATA; ODD-EVEN NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANS; PHOSPHORUS ISOTOPES; PHOSPHORUS-GROUP TRANSFERASES; POTASSIUM ISOTOPES; REACTION KINETICS; RESONANCE; RODENTS; STABLE ISOTOPES; SYNTHESIS; TRANSFERASES; VERTEBRATES; 550500* - Metabolism

Citation Formats

Koretsky, A.P. Phosphorus-31 NMR magnetization transfer measurements of metabolic reaction rates in the rat heart and kidney in vivo. United States: N. p., 1984. Web.
Koretsky, A.P. Phosphorus-31 NMR magnetization transfer measurements of metabolic reaction rates in the rat heart and kidney in vivo. United States.
Koretsky, A.P. 1984. "Phosphorus-31 NMR magnetization transfer measurements of metabolic reaction rates in the rat heart and kidney in vivo". United States. doi:.
@article{osti_6451478,
title = {Phosphorus-31 NMR magnetization transfer measurements of metabolic reaction rates in the rat heart and kidney in vivo},
author = {Koretsky, A.P.},
abstractNote = {This dissertation is concerned with the measurement of the rates of ATP synthesis in the rat kidney and of the creatine kinase catalyzed reaction in the rat heart in situ. Chronically implanted detection coils, employing a balanced matching configuration of capacitors in the tuned circuit, were used to obtain /sup 31/P NMR spectra from heart, kidney, and liver in situ. Gated spectra of heart obtained at systole and diastole and the effects of fructose on kidney and liver were studied. The ability to observe other nuclei using implanted coils is illustrated with /sup 39/K NMR spectra from kidney and muscle. The theoretical considerations of applying magnetization transfer techniques to intact organs are discussed with emphasis on the problems associated with multiple exchange reactions and compartmentation of reactants. Experimental measurements of the ATP synthesis rate (13 ..mu..mol/min/gm tissue) were compared to whole kidney oxygen consumption and Na/sup +/ reabsorption rates to derive ATP/O (0.8 to 1.7) and Na/sup +//ATP (4 to 10) values. The problems associated with ATP synthesis rate measurements in kidney, e.g., the heterogeneity of the inorganic phosphate resonance, are discussed and experiments to overcome these problems proposed.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = 1984,
month = 8
}

Technical Report:
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  • /sup 31/P NMR is a unique tool to study bioenergetics in living cells. The application of magnetization transfer techniques to the measurement of steady-state enzyme reaction rates provides a new approach to understanding the regulation of high energy phosphate metabolism. This dissertation is concerned with the measurement of the rates of ATP synthesis in the rat kidney and of the creatine kinase catalyzed reaction in the rat heart in situ. The theoretical considerations of applying magnetization transfer techniques to intact organs are discussed with emphasis on the problems associated with multiple exchange reactions and compartmentation of reactants. Experimental measurements ofmore » the ATP synthesis rate were compared to whole kidney oxygen consumption and Na/sup +/ reabsorption rates to derive ATP/O values. The problems associated with ATP synthesis rate measurements in kidney, e.g. the heterogeneity of the inorganic phosphate resonance, are discussed and experiments to overcome these problems proposed. In heart, the forward rate through creatine kinase was measured to be larger than the reverse rate. To account for the difference in forward and reverse rates a model is proposed based on the compartmentation of a small pool of ATP.« less
  • Phosphorus-31 NMR magnetization-transfer measurement have been used to measure the flux between ATP and inorganic phosphate during steady-state isometric muscle contraction in the rat hind limb in vivo. Steady-state contraction was obtained by supramaximal sciatic nerve stimulation. Increasing the stimulation pulse width from 10 to 90 ms, at a pulse frequency of 1 Hz, or increasing the frequency of a 10-ms pulse from 0.5 to 2 Hz resulted in an increase in the flux which was an approximately linear function of the increase in the tension-time integral. The flux showed an approximately linear dependence on the calculated free cytosolic ADPmore » concentration up to an ADP concentration of about 90 {mu}M. The data are consistent with control of mitochondrial ATP synthesis by the cytosolic ADP concentration and indicate that the apparent K{sub m} of the mitochondria for ADP is at least 30 {mu}M.« less
  • The reaction of white phosphorus with CCl/sub 4/under the action of light, heat, and gamma radiation was investigated along with the reaction of energy-rich P/sup 32/ recoil atoms with CCl/sub 4/, PCl/sub 3/, CCl/sub 3/PCl/sub 3/, red P, C/sub 2/Cl/sub 6/, and a polymer of chlorine and carbon are produced. In the case of hot-atom experiments only the P/sup 32/-containing compounds were detectable. In the action of gamma radiation on white phosphorus at room temperature, the chief reaction product is red phosphorus, while at 100 deg C practically no more red phosphorus is produced. The reactions appear to consist ofmore » two concurrent processes, one producing red phosphorus and the other PCl/sub 3/ and CCl/sub 3/PCl/sub 2/. The exchange of P/sup 32/ between white phosphorus and PCl/sub 3/ was studied. (M.C.G.)« less
  • In previous reports it was assumed that the energy transmission rate between charged particles was infinite, leading to the assumption that the particles form a Maxwell distribution, Characteristic times for the deuterium plasma are estimated. This estimate shows that in one part of the parameter intervals of main interest for controlled thermonuclear reactions, the assumption of infinite energy transfer rates between the charged particles is a fairly good approximation. In another part the estimate shows that the assumption is not good, but that it is more reasonable to assume three groups of particles, nannely a Maxwell distribution of all electrons,more » a Maxwell distribution of nearly all ions, and a distribution of energetic charged reaction preducts slowing down. Under this assumption parameter sets giving self-sustaining conditiors are found in two cases, namely, when the plasma is fed with pure deuterium and when the plasma is fed with equal amounts of deuterium and tritium. Only under self- sustaining conditions can results from this and the previous reports be directly compared. Comparing csses with equal ion temperatures, changes due to this more realistic assumption are found to be small. It can, however, be seen qualitatively that outside the parameter regions of self-sustaining conditions, changes can be great. (auth)« less