A 1H-nuclear magnetic resonance study on lactate and intracellular pH in frog muscle.

Seo, Yoshiteru; Yoshizaki, Kazuo; Morimoto, Taketoshi

doi:10.2170/jjphysiol.33.721

Cited by 28 publications

(14 citation statements)

References 21 publications

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“…Some of these, such as the higher sensitivity of the 1 H nucleus; the shorter T 1 of the carnosine C-2 proton; more stable concentrations of carnosine throughout rest, exercise, and recovery; the relatively higher frequency range of the pH-induced chemical shift; and narrower linewidth of the C-2 resonance combine to improve temporal resolution and chemical sensitivity. Another advantage of using the C-2 peak to measure pH I is the relative insensitivity of its chemical shift to the concentrations of divalent cations (9,11), particularly in light of the fact that intracellular [Mg 2ϩ ] increases during exercise (e.g., Ref. 35).…”

Section: Discussionmentioning

confidence: 99%

“…Of those studies that used the carnosine C-2 peak to measure pH I in muscle, three employed experimental protocols that might be expected to produce splitting of the carnosine signal. Seo et al (9) stimulated bullfrog femoral biceps muscles to perform three 5-s maximal tetanic contractions, causing the pH I to fall to ϳ6.6 and the [lactate] to rise to ϳ7 mM. The spectra acquired during stimulation indicate no broadening or splitting of the C-2 peak.…”

Section: Why Has This Not Been Detected Before?mentioning

confidence: 99%

“…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.…”

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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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Section: Discussionmentioning

confidence: 99%

Section: Why Has This Not Been Detected Before?mentioning

confidence: 99%

mentioning

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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“…Phosphorous NMR has been used extensively to examine the metabolic profiles in fish muscle during or following exercise or hypoxia exposure (Bock et al, 2002;van den Thillart et al, 1989;van Ginneken et al, 2008;van Ginneken et al, 1995); however, 1 H-NMR has only been used in a handful of studies to examine lactate dynamics in muscle (Seo et al, 1983;Wasser et al, 1992b;Yoshizaki et al, 1981) and fewer studies have used 1 H-NMR in fish (e.g. Bock et al, 2002).…”

Section: H-nmr To Measure Metabolic Effects On Tissuesmentioning

confidence: 99%

Differential recovery from exercise and hypoxia exposure measured using 31P- and 1H-NMR in white muscle of the common carpCyprinus carpio

Hallman

Rojas-Vargas

Jones

et al. 2008

Journal of Experimental Biology

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INTRODUCTIONATP turnover in muscle during high-intensity exhaustive exercise and exposure to hypoxia is primarily supported by substrate-level phosphorylation [creatine phosphate (PCr) hydrolysis and glycolysis], which serves to compensate for the metabolic demands that exceed the capacity of mitochondrial oxidative phosphorylation (Hochachka and Somero, 2002). During hypoxia exposure, hypoxia-tolerant animals, such as the common carp, typically suppress ATP turnover because of an inhibition of mitochondrial function; they rely on substrate-level phosphorylation to maintain energy balance (Bickler and Buck, 2007). By contrast, ATP turnover in white muscle during exhaustive exercise can increase by up to 40-fold (Moyes and West, 1995;Richards et al., 2002a), well beyond the capacity of oxidative phosphorylation to supply ATP and therefore PCr hydrolysis and glycolysis are coordinately increased to support a power output that exceeds oxidative capacity. Although the reasons for the activation of substrate-level phosphorylation during exhaustive exercise or exposure to hypoxia differ, the end metabolic profiles are similar, with low muscle [glycogen] and [PCr], and high [lactate] and metabolic [H + ] (Richards et al., 2007;Wang et al., 1994). In spite of these similar metabolic profiles, no study has directly compared recovery metabolism following exhaustive exercise and exposure to hypoxia.During recovery from exhaustive exercise and hypoxia, pathways must be activated to resynthesize PCr and glycogen. The recovery of these metabolites will be linked because of their dependence on mitochondrial ATP production and metabolic H + use. In particular, the rate of PCr and intracellular pH (pH i ) recovery following exercise or hypoxia exposure will be linked through the creatine kinase catalyzed reaction:where mitochondrial ATP serves as the phosphate donor for free creatine accumulated during the metabolic insult. Phosphocreatine hydrolysis will have an alkalinizing effect on the intracellular fluid, whereas PCr synthesis during recovery will have an acidifying effect. As a result, during recovery there is an almost complete recovery of PCr before pH i and lactate begin to return to normoxic resting levels (Richards et al., 2002b;Schulte et al., 1992;van den Thillart et al., 1989;Wang et al., 1994). Creatine kinase is a near-equilibrium reversible enzyme (Lawson and Veech, 1979) ]. In addition, changes in ATP, ADP free and H + play a dominant role in regulating mitochondrial oxidative phosphorylation and glycogensis, and therefore these metabolites functionally link and coordinate metabolism during recovery.Rates of metabolism and ATP production are strongly influenced by temperature in ectothermic animals, thus temperature acclimation may affect rates of metabolic recovery. Temperature acclimation has been shown to result in dramatic changes in muscle properties that allow many ectothermic fish to maintain activity over a wide range of environmental temperatures (Guderley, 2004). This is in part due to an inverse c...

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“…The changes in phosphorous compounds, lactate, and intracellular pH of muscle due to muscular contraction have been studied in vitro by several investigators using 31P-NMR (BURT et al, 1976;DAWSON et al, 1977DAWSON et al, , 1980YOsHIZAKI, 1978;YAMADA and TANOKURA, 1983), and 1H-NMR (YOSHIZAKI et al, 1981;SEo et al, 1983b). In this report, in vivo study was performed on the aerobic recovery of muscle following tetanus.…”

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In vivo 31P-NMR studies on aerobic recovery of frog muscle following tetanus.

et al. 1984

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Summary Aerobic recovery after a 60-sec tetanus of skeletal muscle of frog was studied using the in vivo 31P-NMR method with a topical magnetic resonance spectrometer (TMR) at 17°C. Creatine phosphate recovered with a time constant of about 25 min-1. Sugar phosphates increased by the Pasteur effect, and saturated at about 4.5 mmol/kg muscle. The high level of the sugar phosphates was sustained during the following 60 min.

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A 1H-nuclear magnetic resonance study on lactate and intracellular pH in frog muscle.

Cited by 28 publications

References 21 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

Differential recovery from exercise and hypoxia exposure measured using 31P- and 1H-NMR in white muscle of the common carpCyprinus carpio

In vivo 31P-NMR studies on aerobic recovery of frog muscle following tetanus.

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