Effects of prior exercise and a low-carbohydrate diet on muscle sarcoplasmic reticulum function during cycling in women

2006 ◽  
Vol 101 (3) ◽  
pp. 695-706 ◽  
Author(s):  
T. A. Duhamel ◽  
H. J. Green ◽  
J. G. Perco ◽  
J. Ouyang

The effects of exercise and diet on sarcoplasmic reticulum Ca2+-cycling properties in female vastus lateralis muscle were investigated in two groups of women following four different conditions. The conditions were 4 days of a low-carbohydrate (Lo CHO) and glycogen-depleting exercise plus a Lo CHO diet (Ex + Lo CHO) ( experiment 2) and 4 days of normal CHO (Norm CHO) and glycogen-depleting exercise plus Norm CHO (Ex + Norm CHO) ( experiment 1). Peak aerobic power (V̇o2peak) was 38.1 ± 1.4 (SE); n = 9 and 35.6 ± 1.4 ml·kg−1·min−1; n = 9, respectively. Sarcoplasmic reticulum properties measured in vitro in homogenates (μmol·g protein−1·min−1) indicated exercise-induced reductions ( P < 0.05) in maximal Ca2+-ATPase activity (0 > 30, 60 min > fatigue), Ca2+ uptake (0 > 30 > 60 min, fatigue), and Ca2+ release, both phase 1 (0, 30 > 60 min, fatigue) and phase 2 (0 > 30, 60 min, fatigue; 30 min > fatigue) in Norm CHO. Exercise was without effect in altering the Hill slope ( nH), defined as the slope of relationship between Ca2+-ATPase activity and Ca2+ concentration. No differences were observed between Norm CHO and Ex+Norm CHO. Compared with Norm CHO, Lo CHO resulted in a lower ( P < 0.05) Ca2+ uptake, phase 1 Ca2+ release (30 min), and nH. Ex + Lo CHO resulted in a greater ( P < 0.05) Ca2+ uptake and nH compared with Lo CHO. The results demonstrate that Lo CHO alone can disrupt SR Ca2+ cycling and that, with the exception of Ca2+ release, a glycogen-depleting session of exercise before Lo CHO can reverse the effects.

2005 ◽  
Vol 99 (3) ◽  
pp. 836-843 ◽  
Author(s):  
G. P. Holloway ◽  
H. J. Green ◽  
T. A. Duhamel ◽  
S. Ferth ◽  
J. W. Moule ◽  
...  

The repetition-dependent effects of a repetitive heavy exercise protocol previously shown to alter muscle mechanic behavior (Green HJ, Duhamel TA, Ferth S, Holloway GP, Thomas MM, Tupling AR, Rich SM, and Yau JE. J Appl Physiol 97: 2166–2175, 2004) on muscle sarcoplasmic reticulum (SR) Ca2+-transport properties, measured in vitro, were examined in 12 untrained volunteers [peak aerobic power (V̇o2 peak) = 44.3 ± 0.66 ml·kg−1·min−1]. The protocol involved 6 min of cycle exercise performed at ∼91% V̇o2 peak once per hour for 16 h. Tissue samples were obtained from the vastus lateralis before (B) and after (A) exercise at repetitions 1 (R1), 2 (R2), 9 (R9), and 16 (R16). Reductions ( P < 0.05) in maximal Ca2+-ATPase activity ( Vmax) of 26 and 12% with exercise were only observed at R1 and R16, respectively. Vmax remained depressed ( P < 0.05) at R2 (B) but not at R9 (B) and R16 (B). No changes were observed in two other kinetic properties of the enzyme, namely the Hill coefficient (defined as the slope of the relationship between Ca2+-ATPase activity and free Ca2+ concentration) and the Ca50 (defined as the free Ca2+ concentration needed to elicit 50% Vmax). Changes in Ca2+ uptake (measured at 2,000 nM) with exercise and recovery generally paralleled Vmax. The apparent coupling ratio, defined as the ratio between Ca2+ uptake and Vmax, was unaffected by the intermittent protocol. Reductions ( P < 0.05) in phase 1 Ca2+ release (32%) were only observed at R1. No differences were observed between B and A for R2, R9, and R16 or between B and B for R1, R2, R9, and R16. The changes in phase 2 Ca2+ release were as observed for phase 1 Ca2+ release. It is concluded that the SR Ca2+-handling properties, in general, display rapid adaptations to repetitive exercise.


2006 ◽  
Vol 291 (4) ◽  
pp. C607-C617 ◽  
Author(s):  
T. A. Duhamel ◽  
H. J. Green ◽  
J. G. Perco ◽  
J. Ouyang

We employed a glycogen-depleting session of exercise followed by a low-carbohydrate (CHO) diet to investigate modifications that occur in muscle sarcoplasmic reticulum (SR) Ca2+-cycling properties compared with low-CHO diet alone. SR properties were assessed in nine untrained males [peak aerobic power (V̇o2 peak) = 43.6 ± 2.6 (SE) ml·kg−1·min−1] during prolonged cycle exercise to fatigue performed at ∼58% V̇o2 peak after 4 days of low-CHO diet (Lo CHO) and after glycogen-depleting exercise plus 4 days of low-CHO (Ex+Lo CHO). Compared with Lo CHO, Ex+Lo CHO resulted in 12% lower ( P < 0.05) resting maximal Ca2+-ATPase activity ( Vmax = 174 ± 12 vs. 153 ± 10 μmol·g protein−1·min−1) and smaller reduction in Vmax induced during exercise. A similar effect was observed for Ca2+ uptake. The Hill coefficient, defined as slope of the relationship between cytosolic free Ca2+ concentration and Ca2+-ATPase activity, was higher ( P < 0.05) at rest (2.07 ± 0.15 vs. 1.90 ± 0.10) with Ex+Lo CHO, an effect that persisted throughout the exercise. The coupling ratio, defined as the ratio of Ca2+ uptake to Vmax, was 23–30% elevated ( P < 0.05) at rest and during the first 60 min of exercise with Ex+Lo CHO. The ∼27 and 34% reductions ( P < 0.05) in phase 1 and phase 2 Ca2+ release, respectively, observed during exercise with Lo CHO were not altered by Ex+Lo CHO. These results indicate that when prolonged exercise precedes a short-term Lo CHO diet, Ca2+ sequestration properties and efficiency are improved compared with those during Lo CHO alone.


2002 ◽  
Vol 283 (4) ◽  
pp. E722-E728 ◽  
Author(s):  
J. D. Schertzer ◽  
H. J. Green ◽  
A. R. Tupling

To examine the thermal instability and the role of sulfhydryl (SH) oxidation on sarcoplasmic reticulum (SR) Ca2+-ATPase function, crude homogenates were prepared from the white portion of the gastrocnemius (WG) adult rat muscles ( n = 9) and incubated in vitro for ≤60 min either at a normal resting body temperature (37°C) or at a temperature indicative of exercise-induced hyperthermia (41°C) with DTT and without DTT (CON). In general, treatment with DTT resulted in higher Ca2+-ATPase and Ca2+ uptake values (nmol · mg protein−1 · min−1, P < 0.05), an effect that was not specific to time of incubation. Incubations at 41°C resulted in lower ( P< 0.05) Ca2+ uptake rates (156 ± 18 and 35.9 ± 3.3) compared with 37°C (570 ± 54 and 364 ± 26) at 30 and 60 min, respectively. At 37°C, ryanodine (300 μM), which was used to block Ca2+ release from the calcium release channel, prevented the time-dependent decrease in Ca2+ uptake. A general inactivation ( P < 0.05) of maximal Ca2+-ATPase activity ( V max) in CON was observed with incubation time (0 > 30 > 60 min), with the effect being more pronounced ( P < 0.05) at 41°C compared with 37°C. The Hill slope, a measure of co-operativity, and the pCa50, the cytosolic Ca2+ concentration required for half-maximal activation of Ca2+-ATPase activity, decreased ( P < 0.05) at 41°C only. Treatment with DTT attenuated the alterations in enzyme kinetics. The increase in V max with the Ca2+ionophore A-23187 was less pronounced at 41°C compared with 37°C. It is concluded that exposure of homogenates to a temperature typically experienced in exercise results in a reduction in the coupling ratio, which is mediated primarily by lower Ca2+ uptake and occurs as a result of increases in membrane permeability to Ca2+. Moreover, the decreases in Ca2+-ATPase kinetics in WG with sustained heat stress result from SH oxidation.


2007 ◽  
Vol 103 (6) ◽  
pp. 1986-1998 ◽  
Author(s):  
T. A. Duhamel ◽  
H. J. Green ◽  
R. D. Stewart ◽  
K. P. Foley ◽  
I. C. Smith ◽  
...  

This study investigated the effects of prolonged exercise, with and without glucose supplementation, on metabolism and sarcoplasmic reticulum (SR) Ca2+-handling properties in working vastus lateralis muscle. Fifteen untrained volunteers [peak O2consumption (V̇o2peak) = 3.45 ± 0.17 l/min; mean ± SE] cycled at ∼60% V̇o2peakon two occasions, during which they were provided with either an artificially sweetened placebo beverage (NG) or a 6% glucose (G) beverage (∼1.00 g carbohydrate/kg body mass). Beverage supplementation started at 30 min of exercise and continued every 15 min thereafter. SR Ca2+handling, metabolic, and substrate responses were assessed in tissue extracted from the vastus lateralis at rest, after 30 min and 90 min of exercise, and at fatigue in both conditions. Plasma glucose during G was 15–23% higher ( P < 0.05) than those observed during NG following 60 min of exercise until fatigue. Cycle time to fatigue was increased ( P < 0.05) by ∼19% during G (137 ± 7 min) compared with NG (115 ± 6 min). Prolonged exercise reduced ( P < 0.05) maximal Ca2+-ATPase activity (−18.4%), SR Ca2+uptake (−27%), and both Phase 1 (−22.2%) and Phase 2 (−34.2%) Ca2+-release rates during NG. The exercise-induced reductions in SR Ca2+-cycling properties were not altered during G. The metabolic responses to exercise were all unaltered by glucose supplementation, since no differences in respiratory exchange ratios, carbohydrate and lipid oxidation rates, and muscle metabolite and glycogen contents were observed between NG and G. These results indicate that the maintenance of blood glucose homeostasis by glucose supplementation is without effect in modifying the muscle metabolic, endogenous glycogen, or SR Ca2+-handling responses.


2005 ◽  
Vol 98 (1) ◽  
pp. 186-192 ◽  
Author(s):  
R. J. Aughey ◽  
C. J. Gore ◽  
A. G. Hahn ◽  
A. P. Garnham ◽  
S. A. Clark ◽  
...  

Athletes commonly attempt to enhance performance by training in normoxia but sleeping in hypoxia [live high and train low (LHTL)]. However, chronic hypoxia reduces muscle Na+-K+-ATPase content, whereas fatiguing contractions reduce Na+-K+-ATPase activity, which each may impair performance. We examined whether LHTL and intense exercise would decrease muscle Na+-K+-ATPase activity and whether these effects would be additive and sufficient to impair performance or plasma K+ regulation. Thirteen subjects were randomly assigned to two fitness-matched groups, LHTL ( n = 6) or control (Con, n = 7). LHTL slept at simulated moderate altitude (3,000 m, inspired O2 fraction = 15.48%) for 23 nights and lived and trained by day under normoxic conditions in Canberra (altitude ∼600 m). Con lived, trained, and slept in normoxia. A standardized incremental exercise test was conducted before and after LHTL. A vastus lateralis muscle biopsy was taken at rest and after exercise, before and after LHTL or Con, and analyzed for maximal Na+-K+-ATPase activity [K+-stimulated 3- O-methylfluorescein phosphatase (3- O-MFPase)] and Na+-K+-ATPase content ([3H]ouabain binding sites). 3- O-MFPase activity was decreased by −2.9 ± 2.6% in LHTL ( P < 0.05) and was depressed immediately after exercise ( P < 0.05) similarly in Con and LHTL (−13.0 ± 3.2 and −11.8 ± 1.5%, respectively). Plasma K+ concentration during exercise was unchanged by LHTL; [3H]ouabain binding was unchanged with LHTL or exercise. Peak oxygen consumption was reduced in LHTL ( P < 0.05) but not in Con, whereas exercise work was unchanged in either group. Thus LHTL had a minor effect on, and incremental exercise reduced, Na+-K+-ATPase activity. However, the small LHTL-induced depression of 3- O-MFPase activity was insufficient to adversely affect either K+ regulation or total work performed.


2004 ◽  
Vol 97 (4) ◽  
pp. 1414-1423 ◽  
Author(s):  
James A. Leppik ◽  
Robert J. Aughey ◽  
Ivan Medved ◽  
Ian Fairweather ◽  
Michael F. Carey ◽  
...  

Prolonged exhaustive submaximal exercise in humans induces marked metabolic changes, but little is known about effects on muscle Na+-K+-ATPase activity and sarcoplasmic reticulum Ca2+ regulation. We therefore investigated whether these processes were impaired during cycling exercise at 74.3 ± 1.2% maximal O2 uptake (mean ± SE) continued until fatigue in eight healthy subjects (maximal O2 uptake of 3.93 ± 0.69 l/min). A vastus lateralis muscle biopsy was taken at rest, at 10 and 45 min of exercise, and at fatigue. Muscle was analyzed for in vitro Na+-K+-ATPase activity [maximal K+-stimulated 3- O-methylfluorescein phosphatase (3- O-MFPase) activity], Na+-K+-ATPase content ([3H]ouabain binding sites), sarcoplasmic reticulum Ca2+ release rate induced by 4 chloro- m-cresol, and Ca2+ uptake rate. Cycling time to fatigue was 72.18 ± 6.46 min. Muscle 3- O-MFPase activity (nmol·min−1·g protein−1) fell from rest by 6.6 ± 2.1% at 10 min ( P < 0.05), by 10.7 ± 2.3% at 45 min ( P < 0.01), and by 12.6 ± 1.6% at fatigue ( P < 0.01), whereas 3[H]ouabain binding site content was unchanged. Ca2+ release (mmol·min−1·g protein−1) declined from rest by 10.0 ± 3.8% at 45 min ( P < 0.05) and by 17.9 ± 4.1% at fatigue ( P < 0.01), whereas Ca2+ uptake rate fell from rest by 23.8 ± 12.2% at fatigue ( P = 0.05). However, the decline in muscle 3- O-MFPase activity, Ca2+ uptake, and Ca2+ release were variable and not significantly correlated with time to fatigue. Thus prolonged exhaustive exercise impaired each of the maximal in vitro Na+-K+-ATPase activity, Ca2+ release, and Ca2+ uptake rates. This suggests that acutely downregulated muscle Na+, K+, and Ca2+ transport processes may be important factors in fatigue during prolonged exercise in humans.


2007 ◽  
Vol 293 (1) ◽  
pp. R354-R362 ◽  
Author(s):  
H. J. Green ◽  
T. A. Duhamel ◽  
K. P. Foley ◽  
J. Ouyang ◽  
I. C. Smith ◽  
...  

Regulation of maximal Na+-K+-ATPase activity in vastus lateralis muscle was investigated in response to prolonged exercise with (G) and without (NG) oral glucose supplements. Fifteen untrained volunteers (14 males and 1 female) with a peak aerobic power (V̇o2peak) of 44.8 ± 1.9 ml·kg−1·min−1; mean ± SE cycled at ∼57% V̇o2peak to fatigue during both NG (artificial sweeteners) and G (6.13 ± 0.09% glucose) in randomized order. Consumption of beverage began at 30 min and continued every 15 min until fatigue. Time to fatigue was increased ( P < 0.05) in G compared with NG (137 ± 7 vs. 115 ± 6 min). Maximal Na+-K+-ATPase activity (Vmax) as measured by the 3- O-methylfluorescein phosphatase assay (nmol·mg−1·h−1) was not different between conditions prior to exercise (85.2 ± 3.3 or 86.0 ± 3.9), at 30 min (91.4 ± 4.7 vs. 91.9 ± 4.1) and at fatigue (92.8 ± 4.3 vs. 100 ± 5.0) but was higher ( P < 0.05) in G at 90 min (86.7 ± 4.2 vs. 109 ± 4.1). Na+-K+-ATPase content (βmax) measured by the vanadate facilitated [3H]ouabain-binding technique (pmol/g wet wt) although elevated ( P < 0.05) by exercise (0<30, 90, and fatigue) was not different between NG and G. At 60 and 90 min of exercise, blood glucose was higher ( P < 0.05) in G compared with NG. The G condition also resulted in higher ( P < 0.05) serum insulin at similar time points to glucose and lower ( P < 0.05) plasma epinephrine and norepinephrine at 90 min of exercise and at fatigue. These results suggest that G results in an increase in Vmax by mechanisms that are unclear.


2001 ◽  
Vol 281 (6) ◽  
pp. E1151-E1158 ◽  
Author(s):  
Sandra J. Peters ◽  
Robert A. Harris ◽  
Pengfei Wu ◽  
Tanya L. Pehleman ◽  
George J. F. Heigenhauser ◽  
...  

The increase in skeletal muscle pyruvate dehydrogenase kinase (PDK) activity was measured in skeletal muscle of six healthy males after a eucaloric high-fat/low-carbohydrate (HF/LC; 5% carbohydrate, 73% fat, and 22% protein of total energy intake) diet compared with a standardized prediet (50% carbohdyrate, 30% fat, and 21% protein). Biopsies were obtained from the vastus lateralis muscle after 3 days on the prediet ( day 0) and after 1, 2, and 3 days of the HF/LC diet. Intact mitchondria were extracted from fresh muscle and analyzed for PDK activity and Western blotting of PDK2 and PDK4 protein. A second biopsy was taken at each time point and frozen for Northern blot analysis of PDK2 and PDK4 mRNAs. PDK activity increased in a linear fashion over the 3-day HF/LC diet and was significantly higher than control by 1 day. PDK activity was 0.09 ± 0.03, 0.18 ± 0.05, 0.30 ± 0.07, and 0.37 ± 0.09 min−1 at 0, 1, 2, and 3 days, respectively. PDK4 protein and mRNA increased maximally by day 1, and PDK2 protein and mRNA were unaffected by the HF/LC diet. Resting respiratory exchange ratios decreased after 1 day of the HF/LC diet (from 0.79 ± 0.02 to 0.72 ± 0.02) and remained depressed throughout the 3-day dietary intervention (0.68 ± 0.01). The immediate shift to fat utilization was accompanied by increased blood glycerol, β-hydroxybutyrate, and plasma free fatty acid concentrations. These results suggest that the continuing increase in PDK activity over the 3-day HF/LC diet is not due to increasing PDK protein beyond 1 day. This could be due to the contribution of another isoform to the total PDK activity or to a continual increase in PDK4 or PDK2 specific activity.


2006 ◽  
Vol 291 (4) ◽  
pp. R1100-R1110 ◽  
Author(s):  
T. A. Duhamel ◽  
J. G. Perco ◽  
H. J. Green

The hypothesis tested was that disturbances in the sarcoplasmic reticulum (SR) Ca2+-cycling responses to exercise would associate with muscle glycogen reserves. Ten untrained males [peak O2consumption (V̇o2 peak) = 3.41 ± 0.20 (SE) l/min] performed a standardized cycle test (∼70% V̇o2 peak) on two occasions, namely, following 4 days of a high (Hi CHO)- and 4 days of a low (Lo CHO)-carbohydrate diet. Both Hi CHO and Lo CHO were preceded by a session of prolonged exercise designed to deplete muscle glycogen. SR Ca2+cycling in crude homogenates prepared from vastus lateralis samples indicated higher ( P < 0.05) Ca2+uptake (μM·g protein−1·min−1) in Hi CHO compared with Lo CHO at 30 min (2.93 ± 0.10 vs. 2.23 ± 0.12) and at 67 min (2.77 ± 0.16 vs. 2.10 ± 0.12) of exercise, the point of fatigue in Lo CHO. Similar effects ( P < 0.05) were noted between conditions for maximal Ca2+-ATPase (μM·g protein−1·min−1) at 30 min (142 ± 8.5 vs. 107 ± 5.0) and at 67 min (130 ± 4.5 vs. 101 ± 4.7). Both phase 1 and phase 2 Ca2+release were 23 and 37% higher ( P < 0.05) at 30 min of exercise and 15 and 34% higher ( P < 0.05), at 67 min during Hi CHO compared with Lo CHO, respectively. No differences between conditions were observed at rest for any of these SR properties. Total muscle glycogen (mmol glucosyl units/kg dry wt) was higher ( P < 0.05) in Hi CHO compared with Lo CHO at rest (+36%), 30 min (+53%), and at 67 min (+44%) of cycling. These results indicate that exercise-induced reductions in SR Ca2+-cycling properties occur earlier in exercise during low glycogen states compared with high glycogen states.


2008 ◽  
Vol 18 (4) ◽  
pp. 399-411 ◽  
Author(s):  
Takaaki Mishima ◽  
Takashi Yamada ◽  
Makoto Sakamoto ◽  
Minako Sugiyama ◽  
Satoshi Matsunaga ◽  
...  

This study was conducted to determine whether dietary chicken-breast extract (CBEX), a rich source of histidine-containing dipeptides, could modify exerciseinduced changes in sarcoplasmic reticulum (SR) function. After 5 weeks of dietary CBEX, SR Ca2+-handling ability was examined in the vastus lateralis muscles of rats subjected to high-intensity running for 2.5 min. Dietary CBEX caused an approximately 15% and 45% increase (p < .01) in muscle carnosine and anserine concentrations, respectively. In resting muscles, depressions in SR Ca2+–ATPase activity were evoked by dietary CBEX without concomitant changes in SR Ca2+ uptake and release rates. The data confirm that high-intensity exercise depresses SR Ca2+ handling. In spite of the same run time, SR Ca2+ handling was reduced to a lesser degree in muscles of CBEX-containing-chow-fed rats than in standard-chow-fed rats (p < .05). These results suggest that dietary CBEX might attenuate deteriorations in SR Ca2+-handling ability that occur with high-intensity exercise.


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