Application of 31P-NMR spectroscopy to the study of striated muscle metabolism

Meyer, Ronald A.; Kuchmerick, M. J.; Brown, Truman R.

doi:10.1152/ajpcell.1982.242.1.c1

Cited by 300 publications

(111 citation statements)

References 35 publications

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“…1 and 2 to Eqs. [1] and [2]. For both the C-2 proton and P i , the pK A is temperaturedependent, becoming higher at lower temperatures ( Table 1).…”

Section: Solutionsmentioning

confidence: 98%

“…pH was calculated using Eq. [1] and the coefficients reported in Results for a temperature of 14°C. Chemical shifts were determined by Lorentzian fitting of the peaks.…”

Section: H Spectroscopymentioning

confidence: 99%

“…The chemical shifts of the C-2 and P i peaks were measured by Lorentzian peak fitting and fit to Eq. [1] or Eq. [2] using Sigma Plot v. 4.01 :…”

Section: Calibration Curvesmentioning

confidence: 99%

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The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

Damon

Hsu

Stark

et al. 2003

Magnetic Resonance in Med

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The appearance of new peaks in the 7.7-8.6 and 6.8 -7.4 ppm regions of the postexercise 1 H spectrum of frog muscle is reported. These new peaks result from the splitting of single pre-exercise carnosine C-2 and C-4 peaks into two peaks, representing the intracellular pH (pH I ) of oxidative and glycolytic fibers. The following data support this conclusion: 1) comparison of means and regression analysis indicates equivalence of the pH I measurements by 1 H and 31 P NMR; 2) the pre-and poststimulation concentrations of carnosine are equal; 3) in ischemic rat hindlimb muscles, the presence of a single, more acidic peak in the plantaris; a single, less acidic peak in the soleus; and two peaks (more and less acidic) in the gastrocnemius correspond to published values for the fiber-type composition of these muscles; and 4) in muscles treated with iodoacetate prior to and during stimulation, a second peak never appears. These data indicate that it is feasible to measure separately the pH I of oxidative and glycolytic fibers using 1 H NMR spectroscopy. Measuring intracellular pH (pH I ) is important in studies of many cellular processes. In studies of muscle metabolism and exercise, it is desirable to measure separately the pH I of oxidative and glycolytic fibers. pH I measurements are commonly made using 31 P NMR spectroscopy, using the inorganic phosphate (P i ) peak's chemical shift. Following intense muscle exercise, the single pre-exercise P i peak frequently splits into two or more poststimulation P i peaks (P i,A and P i,B ), which are believed to correspond to P i in glycolytic and oxidative fibers, respectively (1-7). However, because of the low NMR sensitivity of the phosphorus nucleus, it is sometimes not possible to distinguish a P i peak from noise when the concentration of P i is low. Extreme broadening of the P i peak during exercise has also been observed (5), interfering with the ability to resolve individual peaks.The C-2 and C-4 protons on the imidazole ring of carnosine (␤-alanyl histidine) have pK A 's of approximately 7.0 and chemical shifts that are sensitive to pH in the physiological range, providing an alternative means of measuring pH noninvasively (8). Several authors have taken advantage of this pH sensitivity to measure the pH I of muscle in vivo and ex vivo (8 -13), and of muscle extracts (8,14). Specific applications have been made to muscle in resting (8,10,11,(13)(14)(15)(16), diseased (16), caffeinetreated (14), and exercised (9,11,12) states. Pan et al. (11) and Hitzig et al. (13) found high correlations between pH I measurements by the C-2 and P i methods.While studying exercising ex vivo frog gastrocnemii, we also observed the C-2 peak of carnosine at ϳ8.0 ppm prior to stimulation. However, we noted two peaks (one at ϳ8.53 and one at ϳ8.37 ppm) following exercise. In this study, we tested the hypothesis that they both originate from C-2 protons of carnosine. We also tested the hypothesis that the downfield peak (which we call carnosine C-2 A ) originates from a more acidic compartm...

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“…1 and 2 to Eqs. [1] and [2]. For both the C-2 proton and P i , the pK A is temperaturedependent, becoming higher at lower temperatures ( Table 1).…”

Section: Solutionsmentioning

confidence: 98%

“…pH was calculated using Eq. [1] and the coefficients reported in Results for a temperature of 14°C. Chemical shifts were determined by Lorentzian fitting of the peaks.…”

Section: H Spectroscopymentioning

confidence: 99%

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The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

Damon

Hsu

Stark

et al. 2003

Magnetic Resonance in Med

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“…Since quantitative analyses by both invasive methods and 31 P MRS result in similar skeletal muscle contents for ATP, it is assumed that this compound is fully MRS visible in this tissue (Gadian, 1995). However, 31 P MRS often gives a relatively higher level for PCr, and the difference has been attributed to the rapid breakdown mediated by the creatine kinase reaction during the freezeclamping procedure (Meyer et al 1982); therefore, the PCr level is also considered to be fully MRS visible. On the other hand, 31 P MRS often gives lower levels for Pi than invasive methods, and therefore it is assumed that a significant proportion of this compound is immobilized in subcellular structures or bound to macromolecules (Gadian, 1995).…”

Section: Sensitivity and Magnetic Resonance Visibility In The Detectimentioning

confidence: 99%

“…From a series of such experiments it is possible to estimate the rate-constants of the creatine kinase reaction, and thus the flux through this reaction. Reaction rates of between about 0·05 and 10/s can be estimated by this method (Meyer, 1982;Rudin & Sauter, 1992). We also performed a saturation transfer experiment using mouse skeletal muscle deficient in creatine kinase (see Fig.…”

Section: Muscle Bioenergetics As Observed By Dynamic 31 P Magnetic Rementioning

confidence: 99%

Introduction toin vivo³¹P magnetic resonance spectroscopy of (human) skeletal muscle

et al. 1999

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fax +31 24 3540 866, email a.heerschap@rdiag.azn.nl 31 P magnetic resonance spectroscopy (MRS) offers a unique non-invasive window on energy metabolism in skeletal muscle, with possibilities for longitudinal studies and of obtaining important bioenergetic data continuously and with sufficient time resolution during muscle exercise. The present paper provides an introductory overview of the current status of in vivo 31 P MRS of skeletal muscle, focusing on human applications, but with some illustrative examples from studies on transgenic mice. Topics which are described in the present paper are the information content of the 31 P magnetic resonance spectrum of skeletal muscle, some practical issues in the performance of this MRS methodology, related muscle biochemistry and the validity of interpreting results in terms of biochemical processes, the possibility of investigating reaction kinetics in vivo and some indications for fibre-type heterogeneity as seen in spectra obtained during exercise. P magnetic resonance spectroscopy: Skeletal muscle: High-energy phosphates: Energy metabolism: ExerciseFollowing initial experiments on animal tissue (Hoult et al. 1974;Ackerman et al. 1980), magnetic resonance (MR) spectroscopy (MRS) was first applied to human subjects in the early 1980s, using the 31 P nucleus to monitor the levels and fate of high-energy phosphates in skeletal muscle (Chance et al. 1981;Cresshull et al. 1981;Ross et al. 1981). From these first experiments it was clear that 31 P MRS offers a unique non-invasive window on energy metabolism in skeletal muscle. Of particular interest is the possibility of obtaining important bioenergetic data continuously and with sufficient time resolution during muscle exercise. Another important aspect is that longitudinal monitoring is possible. Numerous studies applying this technique to human subjects have been published, and several reviews are available addressing specific results obtained in this way (for example, see Barbiroli, 1992;Cozzone & Bendahan, 1994;Kemp & Radda, 1994;McCully et al. 1994;Radda et al. 1995).The present paper provides an introduction to 31 P MRS as applied to skeletal muscle of human subjects, and also gives some illustrative examples from our recent studies on skeletal muscle of transgenic mouse models lacking creatine kinase (EC 2.7.3.2). Information content of the 31 P magnetic resonance spectrum of skeletal muscleTo appreciate the potential of the method, first, the information content of a spectrum obtained from human skeletal muscle at rest should be examined (see Fig. 1(A)).The most dominant signals in the spectrum are from phosphocreatine (PCr) and the three non-equivalent phosphate groups of ATP. Usually also a signal for inorganic phosphate (Pi) can be observed, and under favourable conditions signals for phosphomonoesters and phosphodiesters are observable as well. What is the origin of the distinct resonance frequencies of the 31 P nuclei in these compounds? In a first approximation the resonance frequency of the nuclear spins is...

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Direct measurement of high-energy phosphate compounds in patients with neuromuscular disease

1999

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Several neuromuscular disorders are associated with reductions in intramuscular adenosine triphosphate (ATP) and/or phosphocreatine (PCr). These alterations have been primarily characterized using 31P–magnetic resonance spectroscopy (31P‐MRS). We prospectively measured total creatine, PCr, and ATP in muscle biopsies from 81 patients: normal controls (n = 33), mitochondrial cytopathy (n = 8), neuropathic (n = 3), dystrophy/congenital myopathies (n = 7), inflammatory myopathy (n = 12), and miscellaneous myopathies (n = 18) using direct biochemical analysis. Intramuscular concentrations of PCr and ATP were lower for the dystrophy/congenital myopathy, inflammatory myopathy, and mitochondrial disease patients with ragged red fiber (RRF) as compared with normal controls (P < 0.05). Total creatine was lower for the dystrophy/congenital myopathy group as compared with the normal control group (P < 0.05). These values compare favorably to results from other studies using 31P‐MRS and provide external validation for the values obtained using that method. Given the reductions in high‐energy phosphate compounds in these patients, there is the potential for therapeutic intervention with creatine monohydrate supplementation. © 1999 John Wiley & Sons, Inc. Muscle Nerve 22: 1228–1233, 1999.

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Application of 31P-NMR spectroscopy to the study of striated muscle metabolism

Cited by 300 publications

References 35 publications

The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

Introduction toin vivo³¹P magnetic resonance spectroscopy of (human) skeletal muscle

Direct measurement of high-energy phosphate compounds in patients with neuromuscular disease

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Application of 31P-NMR spectroscopy to the study of striated muscle metabolism

Cited by 300 publications

References 35 publications

The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

The carnosine C‐2 proton's chemical shift reports intracellular pH in oxidative and glycolytic muscle fibers

Introduction toin vivo31P magnetic resonance spectroscopy of (human) skeletal muscle

Direct measurement of high-energy phosphate compounds in patients with neuromuscular disease

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Product

Resources

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Introduction toin vivo³¹P magnetic resonance spectroscopy of (human) skeletal muscle