Metabolic rate of oxygen in the skeletal muscle using [15O]O2-PET
Oxygen consumption in skeletal muscle (draft)
Oxygen transport in skeletal muscle is fast. Oxygen may diffuse between paired and crossing microvessels (Kobayashi & Takizawa, 2002), helping to distribute O2 evenly in the tissue, but also contributing to non-nutritive flow. At least in large mammals and in resting muscle, metabolism is independent of O2 transport (Honig et al., 1971).
During single-leg quadriceps exercise, myoglobin saturation drops to ∼50% when at 50-100% of the maximal of quadriceps O2 consumption (Richardson et al., 1995; Richardson et al., 1998; Molé et al., 1999; Sala et al., 2001). Oxygen extraction fraction (OEF), measured from femoral artery sampling, increases from resting value ∼25% to ∼73% during peak exercise (Bangsbo et al., 2000; Sala et al., 2001). The pO2 in muscle capillaries is ∼13 × higher than the intracellular pO2 (Richardson et al., 1998). During moderate exercise, myoglobin saturation stays at a high level (99.6%); hemoglobin saturation is 98% in arterial blood and 69% in venous blood (Mancini et al., 1994); MacDonald et al., 1999). Although perfusion in the muscle increases linearly as a function of contractile force, the oxygen consumption in inactive motor units may remain near the resting level (Lo et al., 2003). Muscle exercise stimulates and maintains glycolysis, independent on of oxygenation state (Conley et al., 1998).
Hartling et al (1989) have measured forearm oxygen uptake, perfusion, and glucose uptake at rest and during maximal exercise using hand ergometer: O2 uptake increased from 6.3±4.1 to 201±56 µmol;/(min*100mL) (∼32-fold), perfusion from 3.6±2.2 to 43±14 mL/(min*100mL) (∼12-fold), and glucose uptake from 0.44±0.07 to 5.7±8.8 µmol/(min*100mL) (∼13-fold).
Steady-state [15O]O2 studies in skeletal muscle
Kairento et al. (1985) have applied steady-state protocol to study perfusion and oxygen consumption in the muscle and soft tissue tumours of rabbits; MRO2 in the muscle was 4.3 ± 2.1 µmol/(min×dL). At steady state, the ratio of the [15O]O2 and [15O]CO2 images, after correction for the concentration of radiowater, was assumed to represent the oxygen extraction fraction (OEF). Mizuno et al. (2003) applied the steady-state inhalation techniques to measure perfusion and OEF in healthy subjects in rest and after exercise; OEF was ∼60% before exercise, and ∼37% after exercise.
OEF has been measured in rabbit muscle infected with Escherichia coli using steady-state [15O]CO2 and [15O]O2; the authors discuss that the OEF estimate may be underestimated, since the extracted [15O]O2 is not instantly converted to radiowater (Senda et al., 1992).
End-capillary pO2 can be estimated based on OEF measured using [15O]O2 PET, and arterial pO2 and pH (Alpert et al., 1988). In this method, the effect of lactate production on end-capillary pH may need to accounted for, if MRO2 and glucose consumption are not coupled.
Single inhalation [15O]O2 bolus model for skeletal muscle
See references Oikonen et al. 1998, Oikonen 1999, and Nuutila et al. 2000.
PET study is performed as described in MET5702.
Steps of MRO2 calculation from single inhalation bolus study:
1. Pre-processing of arterial blood curve
Arterial blood data from the on-line sampler needs to be processed before it can be used as input function in the calculation of oxygen consumption. Also metabolite correction is necessary in muscle studies. Blood TACs are corrected for time delay later by the oxygen model fitting software.
Currently, there is no script for this process. It is strongly suggested that you write the
commands in a batch/script file to pre-process data from
all of your studies.
Detailed instructions can be found in the previous links. Below are the commands needed to
pre-process an imaginary study
us5678, when population averages of whole body oxygen
metabolism are used for metabolite correction:
absscal -c=S:\Lab\plasma\bsampler_calibration\pump_cal.dat -i=O us5678.bld us5678blo.kbq o2metab us5678blo.kbq 1.307e-3 2.034e-3 0 0 tac2svg us5678blo.svg us5678blo_o.kbq us5678blo_w.kbq
After this you would have two data files,
us5678blo_w.kbq, which contain the arterial concentrations of
[15O]O2 and [15O]H2O, respectively.
Both are needed in the analysis.
The corrected blood TAC should always be plotted and controlled visually. It often contains close-to-zero values in the end, which should be removed with a text editor, or left out when determining the fit time. Check also the data units, especially that sample times are in seconds, not minutes. In the example above the graph was written in SVG file.
2. Make regional time-activity curves
Draw the ROIs and compute regional TACs as usual.
Check that data units are the same in regional and blood TACs.
3. Get arterial O2 content ([O2]a)
Arterial oxygen concentration ([O2]a) is not measured in PET Centre blood laboratory but in the hospital laboratory. It is reported in units mL O2/L blood (around 180-200 mL/L). Convert it to units mL O2/dL blood (1 dL = 100 mL = 0.1 L; about 18-20 mL/dL).
If necessary, the concentrations of O2 can be converted between volume and molar units with the molar volume of an ideal gas, 22.4 mL/mmol. Thus, [O2]a in molar units should be about 0.9 mmol O2/100 mL blood.
4. Compute regional MRO2 and OEF values
Estimate the regional MRO2 and OEF (oxygen extraction fraction, or ratio, OER) using fit_mo2, version 3.0.0 or later. This program corrects also the input time delay separately for each region, because there is often marked differences in the tracer appearance times between different muscle regions.
Continuing with the previous example, if the measured arterial oxygen concentration for this subject was 19.8 mL/100 mL:
fit_mo2 -co2a=19.8 -svg=us5678fit.svg us5678blo_o.kbq us5678blo_w.kbq us5678.dft 400 us5678mro2.res
Estimated OEF should be between 0.2 and 0.9. If OEF is too close to 0 or 1, you can try to
- check from regional TACs if there is a probable movement artefact; you may need to draw separate ROIs for different time frame sequences and combine the TACs later, or delete one or two time frames from the regional data
- shorten the estimation time
- redraw the ROIs to make them either smaller (to reduce heterogeneity and movement artefacts) or larger (to reduce noise).
In the above example, the units of MRO2 are ml O2 / (min * 100 ml tissue). If MRO2 is required per tissue mass instead of volume, it can be divided by tissue density (specific gravity), 1.04 g/mL (Reference Man).
- Pre-processing of arterial on-line blood sampler data
- Metabolite correction for [15O]O2
- PET imaging of skeletal muscle
MET5702: PET Centre Intranet > Quality Documents > SOPs and METs > 6 kliininen > menetelmäohjeet (met) > tieteelliset pet-tutkimukset > met5702reisilihasten hapenkulutuksen ([15o]o2) pet-tutkimus valtimoverinäyttein.
Depairon M, Depresseux JC, Zicot M. Quantitation of regional muscle blood flow and oxygen uptake rate at rest and after exercise in arteriopathic patients. Clin Hemorheol. 1988; 8(3-4): 385-390. doi: 10.3233/CH-1988-83-415.
Depairon M, De Landsheere C, Merlo P, Del Fiore G, Quaglia L, Peters JM, Lamotte D, Zicot M. Effect of exercise on the leg distribution of C15O2 and 15O2 in normals and in patients with peripheral ischemia: a study using positron tomography. Int Angiol. 1988; 7(3): 254-257.
Depairon M, Depresseux J-C, Petermans J, Zicot M. Assessment of flow and oxygen delivery to the lower extremity in arterial insufficiency: A PET-scan study comparison with other methods. Angiology 1991; 42: 788-795. doi: 10.1177/000331979104201003.
Depairon M, Zicot M. The quantification of blood flow/metabolism coupling at rest and after exercise in peripheral arterial insufficiency, using PET and 15-O labeled tracers. Angiology 1996; 47(10): 991-999. doi: 10.1177/000331979604701008.
Goldman D. Theoretical models of microvascular oxygen transport to tissue. Microcirculation 2008; 15: 795-811.
Heinonen I, Saltin B, Kemppainen J, Sipilä HT, Oikonen V, Nuutila P, Knuuti J, Kalliokoski K, Hellsten Y. Skeletal muscle blood flow and oxygen uptake at rest and during exercise in humans: a pet study with nitric oxide and cyclooxygenase inhibition. Am J Physiol Heart Circ Physiol. 2011; 300(4): H1510-H1517.
Hällsten K, Yki-Järvinen H, Peltoniemi P, Oikonen V, Takala T, Kemppainen J, Laine H, Bergman J, Bolli GB, Knuuti J, Nuutila P. Insulin- and exercise-stimulated skeletal muscle blood flow and glucose uptake in obese men. Obes Res. 2003; 11(2): 257-265.
ICRP Publication 23, Reference Man: Anatomical, Physiological, and Metabolic Characteristics, International Commission on Radiological Protection, Pergamon Press, New York (1975).
Jones AM, Krustrup P, Wilkerson DP, Berger NJ, Calbet JA, Bangsbo J. Influence of exercise intensity on skeletal muscle blood flow, O2 extraction and O2 uptake on-kinetics. J Physiol. 2012; 590(17): 4363-4376.
Kalliokoski KK, Laaksonen MS, Takala TO, Knuuti J, Nuutila P. Muscle oxygen extraction and perfusion heterogeneity during continuous and intermittent static exercise. J Appl Physiol. 2003; 94(3): 953-958.
Kalliokoski KK, Knuuti J, Nuutila P. Blood transit time heterogeneity is associated to oxygen extraction in exercising human skeletal muscle. Microvasc Res. 2004; 67(2): 125-132.
Kusters YH, Barrett EJ. Muscle microvasculature’s structural and functional specializations facilitate muscle metabolism. Am J Physiol Endocrinol Metab. 2016; 310(6): E379-E387.
Laaksonen MS, Björklund G, Heinonen I, Kemppainen J, Knuuti J, Kyröläinen H, Kalliokoski KK. Perfusion heterogeneity does not explain excess muscle oxygen uptake during variable intensity exercise. Clin Physiol Funct Imaging. 2010; 30(4): 241-249.
Mizuno M, Kimura Y, Iwakawa T, Oda K, Ishii K, Ishiwata K, Nakamura Y, Muraoka I. Regional differences in blood flow and oxygen consumption in resting muscle and their relationship during recovery from exhaustive exercise. J Appl Physiol. 2003; 95(6): 2204-2210. doi: 10.1152/japplphysiol.00197.2003.
Nuutila P, Peltoniemi P, Oikonen V, Larmola K, Kemppainen J, Takala T, Sipilä H, Oksanen A, Ruotsalainen U, Bolli GB, Yki-Järvinen H. Enhanced stimulation of glucose uptake by insulin increases exercise-stimulated glucose uptake in skeletal muscle in humans: studies using [15O]O2, [15O]H2O, [18F]fluoro-deoxy-glucose, and positron emission tomography. Diabetes 2000; 49:1084-1091.
Oikonen V, Nuutila P, Sipilä H, Tolvanen T, Peltoniemi P, Ruotsalainen U. Quantification of oxygen consumption in skeletal muscle with PET and oxygen-15 bolus. Eur J Nucl Med. 1998; 25: 1151. (poster).
Oikonen V. Modelling of low oxygen consumption. In: J. Knuuti, J. Rinne, P.Tenhonen (eds.), Medical Applications of Cyclotrons VIII. Abstracts of the VIII Symposium on the Medical Applications of Cyclotrons. Annales Universitatis Turkuensis D346:16, 1999.
Thorn CE, Kyte H, Slaff DW, Shore AC. An association between vasomotion and oxygen extraction. Am J Physiol Heart Circ Physiol. 2011; 301(2): H442-H449.
Thorn CE, Shore AC. The role of perfusion in the oxygen extraction capability of skin and skeletal muscle. Am J Physiol Heart Circ Physiol. 2016; 310(10): H1277-H1284.
Wagner PD. Heterogeneity of skeletal muscle perfusion and metabolism. J Appl Physiol. 2003; 95: 2202-2203.
Zeller-Plumhoff B, Roose T, Clough GF, Schneider P. Image-based modelling of skeletal muscle oxygenation. J R Soc Interface 2017; 14: 20160992. doi: 10.1098/rsif.2016.0992.
Created at: 2007-09-20
Updated at: 2018-04-07
Written by: Vesa Oikonen