Biomedical subjects

A N Belcastro

Publications and source records attributed to A N Belcastro.

At least 19 recordsLinked to original sources

PubMed1998

Striated muscle calcium-stimulated cysteine protease (calpain-like) activity promotes myeloperoxidase activity with exercise.

An inflammatory response triggered by neutrophil accumulation into muscle tissue is thought to occur with exercise-induced muscle damage. To investigate the relationship between Ca2+-stimulated proteolysis (calpain-like activity) and neutrophil accumulation [myeloperoxidase (MPO) activity], cardiac and plantaris muscles from rats (n = 10) completing 1 h exercise (25 m/min) were investigated. Exercise promoted increases (P<0.05) in both calpain-like and MPO activities; ranging from 2.79 to 58.9 U/g wet weight (ww) and 0.03 to 4.88 U/g ww respectively. Pearson's correlational analysis (r) on calpain-like and MPO activities for cardiac and plantaris muscle data were 0.97 (P<0.001) and 0.68 (P<0.05) respectively, with a combined r of 0.83 (P<0.001) for both muscles across all conditions. To investigate further the extent to which calpain-like activity may promote neutrophil accumulation, another exercise group (n = 5) was pre-injected with the cysteine protease inhibitor, E64c, 1 h before exercise. Administration of E64c lowered calpain-like and MPO activities by 66% and 56% respectively (average from both muscles). From these results it is concluded that a relationship exists between Ca2+-stimulated proteolysis and neutrophil accumulation into striated muscle with exercise, and that the calpain system is involved in localizing the neutrophilic response with exercise.

Animals↗

PubMed1998

Exercise-induced muscle injury: a calpain hypothesis.

It is well established that periods of increased contractile activity result in significant changes in muscle structure and function. Such morphological changes as sarcomeric Z-line disruption and sarcoplasmic reticulum vacuolization are characteristic of exercise-induced muscle injury. While the precise mechanism(s) underlying the perturbations to muscle following exercise remains to be elucidated, it is clear that disturbances in Ca2+ homeostasis and changes in the rate of protein degradation occur. The resulting elevation in intracellular [Ca2+] activates the non-lysosomal cysteine protease, calpain. Because calpain cleaves a variety of protein substrates including cytoskeletal and myofibrillar proteins, calpain-mediated degradation is thought to contribute to the changes in muscle structure and function that occur immediately following exercise. In addition, calpain activation may trigger the adaptation response to muscle injury. The purpose of this paper is to: (i) review the chemistry of the calpain-calpastatin system; (ii) provide evidence for the involvement of the non-lysosomal, calcium-activated neutral protease (calpain) in the response of skeletal muscle protein breakdown to exercise (calpain hypothesis); and (iii) describe the possible involvement of calpain in the inflammatory and regeneration response to exercise.

Calcium↗

PubMed1998

Effect of energy restriction on muscle function and calcium stimulated protease activity in recreationally active women.

The purpose of this study was to investigate whether changes in substrate oxidation that are caused by energy restriction influenced muscle function and skeletal muscle calcium stimulated protease activity in female athletes. Endurance athletes were randomly assigned to maintenance energy (100% kcal) or energy restricted (75% kcal) diet treatment groups for 14 days while maintaining regular activity. Body weight significantly decreased in the 75% diet group (-1.7 +/- 0.3 kg; p < .05), while fat oxidation increased (p < .05). Minimal changes in quadriceps function (assessed using the Kin/Com isokinetic dynamometer) were observed following diet treatment, except selected loss of muscle function in the 75% diet group at a movement velocity of 120 deg/s. These results suggest that increased fat oxidation that is induced by an acute energy restriction does not promote loss of general muscle function and activation of calcium-sensitive muscle proteases.

Adult↗

PubMed1997

A calcium stimulated cysteine protease involved in isoproterenol induced cardiac hypertrophy.

The purpose of this study was to test the relationship between biochemical and functional changes accompanying beta-agonist induced cardiac hypertrophy and the activation of a calcium stimulated cysteine protease. Because the ultrastructural and ionic changes accompanying beta-agonist induced cardiac hypertrophy are reminiscent of the actions of the calcium activated neutral protease, calpain, it was hypothesized that lowering calpain activity (by the use of an exogenous inhibitor(s)) would reduce the extent of hypertrophy. Rats (275-300 g) were randomly assigned to either a control, beta-agonist (iso) or cysteine protease inhibitor (E64c) group. Isoproterenol administration (1 mg/kg) resulted in changes for ventricular weight to body weight ratio (increases 19%), ventricular [RNA] (increases 105.6%), rate of pressure development (increases 22% for +dP/dt) and maximum developed left ventricular pressure (increases 19%) (p < 0.05) after 3 days. Calpain-like activity (assessed by microplate method) increased by 45% (p < 0.05), while [cAMP] returned to control levels (following a transient rise at 1 day; 606.03 +/- 124.1 pmol/g/wet/wt to 937.9 +/- 225 (p < 0.05)). E64c (administered 1 h prior to iso) reduced the extent of hypertrophy, from 19 to 12%, and prevented the increases in; total [RNA], left ventricular function, the initial [cAMP] increase and calpain-like activity. It is concluded that a calcium stimulated cysteine protease(s), such as calpain, may be involved in the biochemical and functional changes associated with isoproterenol induced cardiac hypertrophy.

Adrenergic beta-Agonists↗

PubMed1997

The effect of diazepan and exercise training on selected biochemical and histochemical properties of rat skeletal muscle.

The effects of chronic diazepam (D) treatment and exercise training on total body mass (TBM), microsomal protein yield (MPY), calcium uptake by fragmented sarcoplasmic reticulum (SR), muscle fibre cross-sectional area, and both PFK and SDH activities were investigated in the tibialis anterior (TA), soleus (Sol), and plantaris (Plt) muscles of 50 male albino Sprague-Dawley rats. Rats were assigned randomly to control (C), sprint-trained (S), or endurance-trained (E) groups. Training was of 12 weeks duration. One-half of each group received daily intraperitoneally D doses of 5 mg kg-1 of TBM. Exercise reduced TBM (p < 0.05); increased the relative BM of the TA (E = 2.02 +/- 0.02, p < 0.01) and Plt (E = 1.15 +/- 0.02, p < 0.01; S = 1.13 +/- 0.03, p < 0.01), as well as the Ca++ uptake of the Sol SR (C = 0.08 +/- 0.02, E = 0.16 +/- 01, p < 0.05). MPY was elevated in S-Sol (C = 1.12 +/- 0.6, S = 1.52 +/- 0.1, p < 0.01). D elevated Sol MPY as well as TA PFK. S-trained animals had lower mean fibre areas than the E-trained (D-treated and untreated) animals. The elevated relative masses of TA and Plt are explained by a decreased TBM with exercise. The increased Ca++ uptake of the Sol indicates that E enhances this function, and the increased MPY probably implies an increased SR. The D could be responsible for the D-elevated Sol MPY as well as the TA PFK. El D did not reduce neuromuscular activity to a level adversely affecting oxidative enzyme activity, but in the case of PFK activity in the TA muscle, such a reduction was evident.

Analysis of Variance↗

PubMed1996

Role of calcium-activated neutral protease (calpain) with diet and exercise.

Although the proteolytic events accompanying acute and chronic perturbations in striated muscle protein turnover remain to be fully elucidated, the purpose of this paper is to (a) review the chemistry of the nonlysosomal calpain-calpastatin system, and (b) provide evidence for the involvement of a nonlysosomal, calcium-activated neutral protease (calpain) in the response of skeletal muscle protein breakdown to altered nutritional status (diet composition; energy restriction) and increased periods of contractile activity (exercise). In reviewing the literature, it is apparent that calpain is involved in the protein catabolism which accompanies alterations in diet composition and/or energy restriction. The precise mechanism of calpain action remains to be elucidated; however, the role of altered metabolic status contributing to calcium imbalances is discussed relative to increasing protein degradation. Hypotheses for further investigation are provided in regard to identifying the targeting of selected proteins (and organelles) for degradation by calpain.

Animals↗

PubMed1996

Heart, liver, and skeletal muscle myeloperoxidase activity during exercise.

The purpose of this study was to determine whether contractile activity associated with running exercise was a prerequisite for neutrophil infiltration into rat tissues. H2O2-dependent myeloperoxidase (MPO) activity for rat (n = 8) liver, heart, and gastrocnemius muscles was assayed after 58 +/- 11 min of running to voluntary exhaustion (25 m/min; 0% grade). MPO activity values measured with 0.6 mM H2O2 were 0.988 +/- 0.331 (SD) U/g (skeletal muscle), 1.563 +/- 0.303 U/g (heart), and 1.652 +/- 0.510 U/g (liver) for control samples, compared with 1.690 +/- 0.321, 3.128 +/- 1.221, and 2.752 +/- 0.437 U/g, respectively, for the exercise group (P < or = 0.05). Kinetic analysis revealed that maximum velocity for all tissues increased as a result of the exercise (P < 0.05). The Michaelis constant (Km) values at rest for all tissues were similar (range 0.53-0.57 mM H2O2; P > or = 0.05). Exercise did not alter the Km values for cardiac and liver samples; however, for skeletal muscle, the Km was 28% lower than control (P < or = 0.05). The results of this study show that, with prolonged running, MPO activity is elevated in most rat tissues and not exclusively in skeletal muscle. Moreover, the metabolic status of the tissues may be an important factor for neutrophil infiltration with exercise and not exclusively the type of muscle contraction, as previously hypothesized.

Animals↗

PubMed1994

Calcium-supported calpain degradation rates for cardiac myofibrils in diabetes. Sulfhydryl and hydrophobic interactions.

OBJECTIVE: The purpose was to investigate the calcium required for calpain-mediated degradation of selected cardiac myofibril proteins modified by diabetes, sulfhydryl (SH) and hydrophobic reagents. METHODS: After 20 weeks of streptozotocin-induced (55 mg.kg-1) diabetes, calcium sensitive calpain (1.5 U.ml-1) degradation rates of purified cardiac myofibrillar proteins (1 mg.ml-1) were measured, in vitro, and compared to degradation rates for N-ethylmaleimide (NEM) and 2-p-toluidinylnapthalene-6-sulfonate (TNS) treated samples. RESULTS: Diabetes (blood glucose of 550 +/- 32 mg.dl-1) reduced the yield of purified myofibrillar protein with minimal change in fibril protein composition. Total SH group reactivities (nmol.mg-1.30min) were 220 +/- 21, 163 +/- 17 and 156 +/- 24 for control, diabetic and NEM-treated (0.5 mM) myofibrils (p < or = 0.05). Calpain degradation rates were faster for all diabetic and SH modified myofibrillar proteins (p < or = 0.05), with a 45 and 35% reduction in the pCa50 for a 37 kDa protein of diabetic and NEM-treated fibril complexes. For control myofibrils, both 100 and 200 uM TNS, reduced calpain degradation rates to a similar extent for all substrate proteins. In contrast, diabetic and NEM-treated samples showed a further reduction in calpain degradation rates with increasing TNS from 100 to 200 uM. CONCLUSION: Our results support the hypothesis that in diabetes the calcium requirements for calpain degradation rates are reduced and dependent upon sulfhydryl group status and Ca(2+)-induced hydrophobic interactions, implicating a 37 kDa myofbillar-complexed protein.

Animals↗

PubMed1994

Diaphragm injury and myofibrillar structure induced by resistive loading.

The purpose of this study was to determine whether ventilatory failure is associated with muscle fiber damage and myofibrillar protein alterations. Ventilatory failure was induced by tightening a polyvinyl band around the trachea of hamsters (TB; n = 14) for 6 days, which resulted in severe respiratory acidosis (PCO2: 97.9 +/- 29.6 vs. 51.6 +/- 19.6 Torr; pH: 7.16 vs. 7.35), hypoxemia (PO2: 42.8 +/- 16.8 vs. 65.9 +/- 25.8 Torr), and increased pulmonary resistance (1.89 +/- 1.61 vs. 0.29 +/- 0.27 cmH2O.ml-1 x min; P < 0.05). The point-counting technique of hematoxylin- and eosin-stained cross sections showed a higher area fraction of abnormal muscle and inflammatory cells in the costal [0.133 +/- (SE) 0.33 vs. 0.040 +/- 0.010] and crural regions (0.069 +/- 0.020 vs. 0.012 +/- 0.003) of the diaphragm in TB hamsters than in control hamsters. Electron micrographs revealed sarcomeric disruption and Z band streaming in the diaphragm of TB hamsters. Myofibrillar changes of the diaphragm associated with ventilatory failure were quantitative (i.e., a lower yield of purified myofibrils) but not qualitative (similar sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles); however, sulfhydryl group reactivities were reduced (P < 0.05). Proteolysis of purified myofibrils from the diaphragm digested with calpain showed faster degradation rates for tropomyosin and alpha-actinin but not for all proteins for the TB animals. Ventilatory failure induced by resistive loading was associated with diaphragm injury; some of this injury was linked to changes in myofibrillar complexes, specifically their susceptibility to calpain-mediated degradation.

Actinin↗

PubMed1993

Circadian rhythm of glycogen in the hamster diaphragm.

The purpose of this study was to determine the diurnal fluctuation of glycogen stores for the whole hemidiaphragm and within a specific myofibrillar ATPase (M-ATPase) fibre type and diaphragmatic region. Fifty-six golden Syrian hamsters were randomly divided into six groups according to the time of sampling biopsies from the diaphragm: 03:00, 07:00, 11:00, 15:00, 19:00, and 23:00. The right hemidiaphragm was quick frozen and biochemically assayed for glycogen levels. Biopsies from the left hemidiaphragm of the same animal were cut from the anterior costal and crural regions, and stained with periodic acid--Schiff (PAS) and for M-ATPase. Optical density measures of PAS-stained fibres were determined to quantitate glycogen in different M-ATPase fibre types and diaphragmatic regions. Biochemical assay of the entire hemidiaphragm showed slightly greater glycogen content of biopsies taken at 11:00 and 15:00 than at 03:00, 19:00, and 23:00 (range of differences: 6.4-10.0%). However, glycogen levels within a specific M-ATPase fibre type and diaphragm region were not different in biopsies sampled at different times. Because the hamster has a small diurnal variation of glycogen in the diaphragm, which is similar to the small diurnal variation of glycogen in human skeletal muscle, this species may be a good animal model for metabolic studies of the diaphragm that could be affected by diurnal glycogen variability.

Adenosine Triphosphatases↗

PubMed1993

Skeletal muscle calcium-activated neutral protease (calpain) with exercise.

The purpose of this study was to investigate whether exercise could induce calpain activation by altering the Ca2+ required for half-maximal activity (pCa50) and/or susceptibility of digestible muscle protein substrates. Rats (225 g) were assigned to control, exercise (25 m/min, 0% grade), and 24-h recovery groups. Exercise resulted in a generalized 48 +/- 18% loss of muscle glycogen and a twofold increase in plasma creatine kinase levels (P < or = 0.05). Exercise increased total caseinolysis of diethylaminoethyl Sepharose-prepared low (u) and high (m) Ca2+ calpain isoforms by 22 and 30%, respectively (P < or = 0.05). The pCa50 of u- and m-calpain with exercise increased from 5.98 +/- 0.12 to 6.20 +/- 0.15 (P > or = 0.05) and from 3.63 +/- 0.10 to 3.90 +/- 0.16 (P > or = 0.05), respectively. In vitro, calpain-mediated degradation/disappearance rates (i.e., percentage of protein degraded in 10 min) for control tropomyosin and alpha-actinin were 69 and 30% compared with 92 and 61% after exercise (P < or = 0.05). The results of this study confirm that level running increases total nonlysosomal Ca2+ specific protease activity, which may promote exercise-induced muscle damage or fatigue.

Animals↗

PubMed1993

Function of skeletal muscle sarcoplasmic reticulum vesicles with exercise.

In this study the response of sarcoplasmic reticulum (SR) to prolonged moderate-intensity exercise was examined in highly purified native vesicles isolated from rat gastrocnemius muscle. Maximal Ca(2+)-dependent ATP hydrolysis was reduced by 12.6% within 2 min after the onset of exercise. The reduction in Ca(2+)-dependent adenosinetriphosphatase activity progressed to 18% at 30 min of exercise and was maintained throughout the subsequent 90-100 min of exercise. Oxalate stimulation of unidirectional Ca2+ transport (Ca2+ loading) was unaffected by exercise. However, in the absence of anion stimulation, steady-state Ca2+ uptake (bidirectional flux) was 51.2 +/- 7.3 nmol Ca2+/mg SR after exercise compared with 36.2 +/- 2.5 nmol Ca2+/mg SR for the control period (P < 0.05). Anion-induced Ca2+ release increased from a control value of 33.9 +/- 4.3 to 55.9 +/- 9.8 nmol Ca2+/mg SR after exercise (P < 0.05). The mechanistic basis for the increase in apparent Ca(2+)-ATP coupling is unclear, although the early onset of the changes suggests the potential for functional adaptation for the SR in response to increased contractile activity.

Animals↗

PubMed1992

Intraluminal Ca2+ dependence of Ca2+ and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum Ca2+ release.

The action of ryanodine upon sarcoplasmic reticulum (SR) Ca2+ handling is controversial with evidence for both activation and inhibition of SR Ca2+ release. In this study, the role of the intraluminal SR Ca2+ load was probed as a potential regulator of ryanodine-mediated effects upon SR Ca2+ release. Through dual-wavelength spectroscopy of Ca2+:antipyrylazo III difference absorbance, the intraluminal Ca2+ dependence of ryanodine and Ca(2+)-induced Ca2+ release (CICR) from skeletal SR vesicles was examined. Ryanodine addition after initiation of Ca2+ uptake (a) increased the intraluminal Ca2+ sensitivity of CICR and (b) stimulated spontaneous Ca2+ release with a delayed onset. These ryanodine effects were inversely proportional to the intraluminal Ca2+ load. Ryanodine also inhibited subsequent CICR after reaccumulation of Ca2+ released from the initial CICR. These results provide evidence that ryanodine inhibits transitions between low and high affinity Ca2+ binding states of an intraluminal Ca2+ compartment, possibly calsequestrin. Conformational transitions of calsequestrin may be reciprocally coupled to transitions between open and closed states of the Ca2+ release channel.

Adenosine Triphosphate↗

PubMed1992

Calcium-activated neutral protease effects upon skeletal muscle sarcoplasmic reticulum protein structure and calcium release.

In this study, the effects of Ca(2+)-activated neutral protease (CANP) upon skeletal muscle heavy sarcoplasmic reticulum (HSR) structure and function were investigated. CANP was immunolocalized to the 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid detergent-insoluble fraction of purified HSR membranes. Ca2+ activation of the endogenous membrane-bound CANP produced a characteristic partial fragmentation of the HSR 565-kDa Ca2+ release channel. Similarly, the major substrate for both micromolar and millimolar Ca(2+)-sensitive isoforms of exogenous CANP was the Ca2+ release channel with proteolysis of a 88-kDa HSR protein also observed. Ca2+ release channel proteolysis was initiated at a single cleavage site with coincidental production of 410- and 150-kDa peptide fragments. Appearance of 160- and 137-kDa limiting peptides accompanied secondary proteolysis of the primary 410- and 150-kDa fragments, respectively. Despite extensive proteolysis of the Ca2+ release channel, CANP did not dramatically alter the Ca2+ handling and ryanodine binding properties of HSR membranes. The association of CANP with isolated HSR membranes suggests that, in vivo, this protease may modify an additional property of the Ca2+ release channel. This may be related to the CANP-susceptible structural association of the Ca2+ release channel with dihydropyridine receptors at T-tubule/sarcoplasmic reticulum junctions.

Adenosine Triphosphate↗

PubMed1992

Effect of manipulation of plasma lactate on integrated EMG during cycling.

This investigation was undertaken to record electromyographic activity of the vastus lateralis muscle during incremental cycling exercise and to determine whether it would be sensitive to altered dynamics of plasma lactate increases seen with intense exercise. Trained cyclists (N = 6) performed two progressive, stepwise exercise tests (23.5 W.min-1) to fatigue on a cycle ergometer at 90 rpm. One of the exercise tests was preceded by arm ergometer exercise in an attempt to elevate the circulating plasma lactate levels prior to starting the criterion exercise test. The starting mean plasma lactate values were 4.59 and 26.69 mmol lactate.-1 for the two exercise sessions. Cardiorespiratory values did not differ significantly between exercise sessions completed in the absence and presence of increased circulating plasma lactate. The no-arm trial (i.e., nonelevated plasma lactate condition) was associated with a plasma lactate inflection point (Tlac) at 72.6% VO2max. Previous arm exercise elevated the lactate such that during the criterion exercise plasma lactate values were decreasing with increasing power output at lower exercise intensities. As exercise intensity increased lactate values also increased beginning at a power output of about 76% VO2 max. Mean per cycle integrated EMG (CIEMG) increased linearly with increased power output in both exercise sessions. The slopes of the EMG-power output curve were not significantly different (P less than 0.05). There were no inflection points in these curves. The absence of an inflection point show that surface EMG does not provide an indication of Tlac.

Adolescent↗

PubMed1991

Regulation of ATP-stimulated releasable myofilaments from cardiac and skeletal muscle myofibrils.

The mechanism underlying the formation of easily releasable myofilaments, from myofibrils treated with an ATP-containing relaxing solution, was examined in this investigation. The proportion of releasable myofilaments purified from myofibrils of cardiac, fast- and slow-twitch muscles increased as the [ATP] was raised from 0 to 8.5 mM. The protein composition of the easily releasable myofilaments did not differ with increasing ATP concentrations as observed by 5-15% linear gradient SDS-PAGE. There is a nucleotide specificity to the release of myofilaments in the order of ATP greater than GTP much greater than UTP greater than CTP. Experiments with AMP-PNP and inorganic phosphate (Pi) showed that ATP hydrolysis and the build up of Pi are not requirements in the formation of the easily releasable myofilaments. The release of myofilaments was found to be insensitive to variations in pH from 6.5 to 7.5. The ATP stimulation of myofilaments release is ubiquitin-independent, since incubation of purified myofibrils with ubiquitin (1-100 micrograms/ml) at both 20 and 37 degrees C did not change the amount released. Modifying the free sulfhydryl group content by treatment of myofibrils with NEM (0.01-1 mM) or silver nitrate (0.1-10 mM) decreased the proportion of myofilaments that were releasable. Exclusion of 1 mM DTT from the preparation of myofibrils had similar results. These results indicate that the formation of easily releasable myofilaments can be mediated by metabolically related parameters such as the adenosine nucleotides and the reduction-oxidation status of the myofibrillar proteins of striated muscle.

Actin Cytoskeleton↗

PubMed1991

Biochemical adaptation of cardiac and skeletal muscle to physical activity.

1. Female Wistar rats were randomly assigned to control (C) or exercising (T) groups and subsequently portioned into 1, 3, 5 and 10 day T and C groups. The T groups completed a progressive endurance running program. Biochemical indices of adaptation were measured in cardiac muscle and in plantaris and soleus muscles of C and T animals after their last exercise bout. 2. In cardiac muscle, myofibrillar ATPase activity was significantly elevated in the 3T (0.241 +/- 0.031) and 5T (0.242 +/- 0.013) groups (P less than or equal to 0.05) compared to their respective controls (3C = 0.187 +/- 0.015 and 5C = 0.190 +/- 0.007). 3. After 10 days of training cardiac myofibrillar ATPase activity was elevated by 17% but this was not significant (P greater than or equal to 0.05). 4. No changes in myofibrillar ATPase activity were seen in skeletal muscle (P greater than or equal to 0.05), however, hexokinase activity progressively increased and was significantly elevated in the 3T, 5T and 10T soleus and plantaris muscles of rats over controls (P less than or equal to 0.05). 5. Minimal nonsignificant changes were noted in the hexokinase activity of the hearts of all T groups (P greater than or equal to 0.05). 6. These results indicate that metabolic adaptation of the heart and skeletal muscles takes place after as little as three training sessions. 7. Although the adaptation of the skeletal muscles continually progresses, the adaptation of the heart appears to be transitory.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗

PubMed1990

Improved resolution of the initial fast phase of heavy sarcoplasmic reticulum Ca2+ uptake by Ca2+:antipyrylazo III dual-wavelength spectroscopy.

The effect of ATP upon difference absorbance due to Ca2+ and Mg2+ complexation with the metallochromic dye, Antipyrylazo III (AP III), was investigated. At divalent cation concentrations appropriate for Sarcoplasmic Reticulum Ca2+ transport, wavelengths (greater than 670 nm) were found whereupon the addition of up to 1mM nucleotide did not alter divalent cation:AP III difference absorbance. At these sample wavelengths an initial rapid uptake of Ca2+ by Heavy SR (HSR) was clearly resolved by dual wavelength spectroscopy of Ca2+:dye difference absorbance. Elimination of ATP interference of Ca2+:AP III absorbance by Mg2+ elevation (3-10mM) was shown to be an inappropriate general strategy for AP III spectroscopic studies of HSR Ca2+ transport due to Mg2+ inhibition of ryanodine receptor mediated Ca2+ release.

Adenosine Triphosphate↗