Subcellular movement and expression of HSP27, αB-crystallin, and HSP70 after two bouts of eccentric exercise in humans (original) (raw)
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AJP: Regulatory, Integrative and Comparative Physiology, 2007
In this study the stress protein response to unaccustomed maximal eccentric exercise in humans was investigated. Eleven healthy males performed 300 maximal eccentric actions with the quadriceps muscle. Biopsies from vastus lateralis were collected at 30 min and 4, 8, 24, 96, and 168 h after exercise. Cellular regulation and localization of heat shock protein (HSP) 27, ␣B-crystallin, and HSP70 were analyzed by immunohistochemistry, ELISA technique, and Western blotting. Additionally, mRNA levels of HSP27, ␣B-crystallin, and HSP70 were quantified by Northern blotting. After exercise (30 min), 81 Ϯ 8% of the myofibers showed strong HSP27 staining (P Ͻ 0.01) that gradually decreased during the following week. ␣B-Crystallin mimicked the changes observed in HSP27. After exercise (30 min), the ELISA analysis showed a 49 Ϯ 13% reduction of the HSP27 level in the cytosolic fraction (P Ͻ 0.01), whereas Western blotting revealed a 15-fold increase of the HSP27 level in the myofibrillar fraction (P Ͻ 0.01). The cytosolic HSP70 level increased to 203 Ϯ 37% of the control level 24 h after exercise (P Ͻ 0.05). After 4 days, myofibrillarbound HSP70 had increased ϳ10-fold (P Ͻ 0.01) and was accompanied by strong staining on cross sections. mRNA levels of HSP27, ␣B-crystallin, and HSP70 were all elevated the first day after exercise (P Ͻ 0.01); HSP70 mRNA showed the largest increase (20-fold at 8 h). HSP27 and ␣B-crystallin seemed to respond immediately to maximal eccentric exercise by binding to cytoskeletal/myofibrillar proteins, probably to function as stabilizers of disrupted myofibrillar structures. Later, mRNA and total HSP protein levels, especially HSP70, increased, indicating that HSPs play a role in skeletal muscle recovery and remodeling/adaptation processes to high-force exercise. stress proteins; muscle damage; eccentric actions; skeletal muscle MYOFIBRILLAR DISORGANIZATION and membrane disruption are hallmarks of exercise-induced muscle damage (24, 32), and the functional outcome of such muscle damage is reduced forcegenerating capacity (55). Subsequent inflammatory events in the exercised muscles, such as infiltration of blood-borne leukocytes, may be necessary for repair, regeneration, and adaptation, but the process itself may be transiently damaging because of release of reactive oxygen species from the infiltrated cells (51, 52).
Journal of Applied Physiology, 2014
Frankenberg NT, Lamb GD, Overgaard K, Murphy RM, Vissing K. Small heat shock proteins translocate to the cytoskeleton in human skeletal muscle following eccentric exercise independently of phosphorylation. Small heat shock proteins (sHSPs) are a subgroup of the highly conserved family of HSPs that are stress inducible and confer resistance to cellular stress and injury. This study aimed to quantitatively examine whether type of contraction (concentric or eccentric) affects sHSPs, HSP27 and ␣B-crystallin, localization, and phosphorylation in human muscle. Vastus lateralis muscle biopsies from 11 healthy male volunteers were obtained pre-and 3 h, 24 h, and 7 days following concentric (CONC), eccentric (ECC1), and repeated bout eccentric (ECC2) exercise. No changes were apparent in a control group (n ϭ 5) who performed no exercise. Eccentric exercise induced muscle damage, as evidenced by increased muscle force loss, perceived muscle soreness, and elevated plasma creatine kinase and myoglobin levels. Total HSP27 and ␣B-crystallin amounts did not change following any type of exercise. Following eccentric exercise (ECC1 and ECC2) phosphorylation of HSP27 at serine 15 (pHSP27-Ser15) was increased approximately 3-to 6-fold at 3 h, and p␣B-crystallin-Ser59 increased ϳ10-fold at 3 h. Prior to exercise most of the sHSP and psHSP pools were present in the cytosolic compartment. Eccentric exercise resulted in partial redistribution of HSP27 (ϳ23%) from the cytosol to the cytoskeletal fraction (ϳ28% for pHSP27-Ser15 and ϳ7% for pHSP27-Ser82), with subsequent full reversal within 24 h. ␣B-crystallin also showed partial redistribution from the cytosolic to cytoskeletal fraction (ϳ18% of total) 3 h post-ECC1, but not after ECC2. There was no redistribution or phosphorylation of sHSPs with CONC. Eccentric exercise results in increased sHSP phosphorylation and translocation to the cytoskeletal fraction, but the sHSP translocation is not dependent on their phosphorylation.
A single bout of eccentric exercise increases HSP27 and HSC/HSP70 in human skeletal muscle
Acta Physiologica Scandinavica, 2001
Changes in heat shock proteins (HSPs), HSP27 and HSC/HSP70 were characterized in human biceps brachii muscle following damaging high-force eccentric exercise. Male and female volunteers performed a maximal eccentric resistance exercise with the elbow flexor muscles of the nondominant arm known to be sufficient to cause substantial muscle damage. Protein extracts of biopsy tissue samples taken 48 h post-exercise were immunoblotted for HSC/HSP70 and HSP27. Densitometric analysis demonstrated that these proteins increased significantly (P < 0.01) in the damaged biceps brachii relative to the control arm. The HSC/HSP70 increased 1064% in the exercised sample while HSP27 increased by 234%. Although the literature reports a muscular heat shock response following aerobic, oxidative exercise, this is the ®rst documentation of increases in protein expression of both HSC/HSP70 and HSP27 in human skeletal muscle in response to a single bout of resistance exercise.
Acta Physiologica, 2009
Aim: This study hypothesized that heat shock protein (HSP) translocation and upregulation is more probable to occur after eccentric exercise than after concentric exercise or repeated eccentric exercise. Methods: Fourteen young, healthy, untrained male subjects completed two bench-stepping exercise bouts with 8 weeks between bouts, and were compared with a control group (n = 6). Muscle biopsies collected from m. vastus lateralis of both legs prior to and at 3 h, 24 h and 7 days after exercise were quantified for mRNA levels and/or for HSP27, ab-crystallin and inducible HSP70 content in cytosolic and cytoskeletal protein fractions. Results: The first bout of exercise reduced muscle strength and increased muscle soreness predominantly in the eccentric leg (P < 0.05). These responses were attenuated after the repeated eccentric exercise bout (P < 0.05), suggesting a repeated bout adaptation. Increases in inducible HSP70 and HSP27 protein content in cytoskeletal fractions were observed exclusively after eccentric exercise (P < 0.05). For HSP27, an approx. 10-fold upregulation after first-bout eccentric exercise was attenuated to a an approximately fourfold upregulation after the repeated eccentric exercise bout. mRNA levels for HSP70, HSP27 and ab-crystallin were upregulated within approximately two to fourfold ranges at time points 3 and 24 h postexercise (P < 0.05). This upregulation was induced exclusively by eccentric exercise but with a tendency to attenuated expression 3 h after the repeated eccentric exercise bout. Conclusion: Our results show that HSP translocation and expression responses are induced by muscle damaging exercise, and suggest that such HSP responses are closely related to the extent of muscle damage.
European Journal of Applied Physiology, 2006
Increased HSP expression in response to acute exercise is well documented in animal studies, and there is growing evidence that similar responses occur in man. In general, many human exercise studies have investigated the HSP response to low force continuous activity, while the knowledge about the HSP response to high force intermittent type of activity, like weight training, is so far sparse. In addition, most studies have used untrained subjects, and a common observation is that acute low force continuous activity in untrained individuals increases the HSP expression in these individuals.The main scope of this study was to investigate the HSP response in very well trained males subjected to longitudinal high intensity exercise, and if this response was dependent on exercise modality [i.e. eccentric (ECC) or concentric (CON) contractions]. Very well trained males performed progressive strength training consisting of either high force ECC or high force CON elbow flexions 2-3 times a week for 12 weeks. Compared with preexercise levels, HSP72 expression decreased by 46.1% (P<0.05) after CON contractions. GRP75 expression was unchanged after ECC or CON contractions, while ubiquitin expression decreased by 19.9% (P<0.02) after ECC contractions. These findings imply that chronic, intensive exercise may attenuate the HSP response in well-trained males.
Induction of heat shock protein 72 mRNA in skeletal muscle by exercise and training
Equine Veterinary Journal, 2010
In response to stress, cells synthesise heat shock proteins (HSP) to maintain protein homeostasis. To study whether exercise and training induce expression of HSP72 in the middle gluteal muscle, 10 Finnhorses performed a submaximal 60 min exercise test on a treadmill. Test A was performed after 3 months of training, and the other two tests 2 (B) and 5 (C) weeks later. Blood samples were taken during and after the tests, and biopsy samples before, immediately after and 23 h after each test. HSP72 mRNA was analysed using a digoxigenin-labelled probe. Blood lactate concentration in the 3 tests varied between 7.2 and 10.2 mmol/l. Training increased HSP72 mRNA, as indicated by increases in samples taken at rest (A<B<C). Exercise also tended to increase HSP72 mRNA transiently but, 23 h later, values had returned to pre-exercise levels. HSP72 mRNA was expressed in all muscle fibres. After exercise, HSP72 mRNA correlated positively with the peak concentration of blood lactate, but not with indicators of energy status. Therefore, acidosis rather than energy depletion was the major inducer of HSP72 expression after moderate intensity exercise. Because HSP72 may protect cells against stress, knowledge about their expression may help in planning optimal trainng regimes.
European Journal of Applied Physiology, 2006
Forty untrained persons were randomized to four diVerent training protocols that exercised the m. triceps brachii. Group 1 and 2 performed high intensity (HI) elbow extensions and group 3 and 4 performed low intensity (LI) elbow extensions. Group 1 and 3 trained until they had accumulated a matching high volume (HV) of training, while group 2 and 4 trained until they had accumulated a matching low volume (LV) of training. Training for 5-8 weeks increased the HSP72, HSP27 and GRP75 levels in the subjects' m. triceps brachii by 111, 71 and 192%, respectively ( . There were, however, no signiWcant diVerences in the heat shock protein (HSP) responses to training between the four training groups . The frequency of extreme responses to exercise was, however, higher after HI exercise than after LI exercise, indicating that HI exercise induces extreme HSP reactions in some subjects. When we assigned the subjects to three clusters, according to the total number of repetitions they had lifted, the subjects who had lifted the highest number of repetitions had lower PostExc HSP levels compared with subjects that lifted the low-est number of repetitions ( . Additionally, there was a negative non-linear regression between the subjects PreExc levels of HSP72, HSP27 and GRP75 and the percentage change in their respective protein concentration after training (r = ¡0.75, ¡0.89 and ¡0.88, all P < 0.0001). Thus, the PreExc level of HSPs seems to be an important "regulator" of HSP expression following the training.
Exercise-induced heat shock protein (HSP70) response in human skeletal muscle and leukocytes
Medicina Sportiva
Heat shock proteins (HSPs) represent a family of highly conserved stress proteins found in all organisms from bacteria to humans. There is also substantial evidence that induction of HSPs synthesis occurs in vivo in response to a variety of physiologically relevant stressors, such as exercise, hyperthermia, oxidative stress, and other metabolic insults. The main aim of this review was to discuss the family of 70kDa heat shock proteins (HSP70), their structure and functions as well as the regulation of their expression in human skeletal muscle and leukocytes in response to exercise.
European Journal of Applied Physiology, 2013
Non-steroidal anti-inflammatory drugs (NSA-IDs) are widely consumed in relation to pain and injuries in skeletal muscle, but may adversely affect muscle adaptation probably via inhibition of prostaglandin synthesis. Induction of heat shock proteins (HSP) represents an important adaptive response in muscle subjected to stress, and in several cell types including cardiac myocytes prostaglandins are important in induction of the HSP response. This study aimed to determine the influence of NSAIDs on the HSP response to eccentric exercise in human skeletal muscle. Healthy males performed 200 maximal eccentric contractions with each leg with intramuscular infusion of the NSAID indomethacin or placebo. Biopsies were obtained from m. vastus lateralis before and after (5, 28 hrs and 8 days) the exercise bout from both legs (NSAID vs unblocked leg) and analysed for expression of the HSPs HSP70, HSP27 and aB-crystallin (mRNA and protein). NSAID did not affect the mRNA expression of any of the HSPs. Compared to pre values, the mRNA expression of all HSPs was increased; aB-crystallin, 3.6-and 5.4-fold; HSP70, 26-and 3.4-fold; and HSP27: 4.8-and 6.5-fold at 5 and 28 hrs post-exercise, respectively (all p \ 0.008). Immunohistochemical stainings for aB-crystallin and HSP70 revealed increased staining in some samples but with no differences between legs. Changes in force-generating capacity correlated with both aB-crystallin and HSP70 mRNA and immunohistochemisty data. Increased expression of HSPs was observed on mRNA and protein level following eccentric exercise; however, this response was unaffected by local intramuscular infusion of NSAIDs Keywords HSP Á Non-steroidal anti-inflammatory drugs Á Muscle damage Á Force recovery Á mRNA Á RT-PCR Á Immunohistochemistry Communicated by David C. Poole.
Biology
Skeletal muscle is a plastic and complex tissue, rich in proteins that are subject to continuous rearrangements. Skeletal muscle homeostasis can be affected by different types of stresses, including physical activity, a physiological stressor able to stimulate a robust increase in different heat shock proteins (HSPs). The modulation of these proteins appears to be fundamental in facilitating the cellular remodeling processes related to the phenomenon of training adaptations such as hypertrophy, increased oxidative capacity, and mitochondrial activity. Among the HSPs, a special attention needs to be devoted to Hsp60 and αB-crystallin (CRYAB), proteins constitutively expressed in the skeletal muscle, where their specific features could be highly relevant in understanding the impact of different volumes of training regimes on myofiber types and in explaining the complex picture of exercise-induced mechanical strain and damaging conditions on fiber population. This knowledge could lead ...