Skip to main content

Advertisement

SpringerLink
Log in

MAPK, androgen, and glucocorticoid receptor phosphorylation following high-frequency resistance exercise non-functional overreaching

  • Original Article
  • Published:
European Journal of Applied Physiology Aims and scope Submit manuscript

Abstract

Purpose

Stressful training with insufficient recovery can impair muscle performance. Expression of mitogen-activated protein kinases (MAPK) has been reported at rest following overreaching and overtraining. The acute myocellular exercise response to stressful training with insufficient recovery has not been investigated. We investigated MAPK, androgen, and glucocorticoid receptor phosphorylation following a period of stressful training.

Methods

Sixteen resistance-trained men were matched on barbell squat 1 repetition maximum strength and randomized into a group that performed normal training or stressful training with insufficient recovery. The control group (CON) performed three speed-squat training sessions on non-consecutive days, while the stressful training group (NFOR) performed 15 training sessions over 7.5 days. Resting and post-exercise skeletal muscle biopsies were obtained prior to (T1) and after the training period (T2). Samples were analyzed for total and phosphorylated androgen receptor (AR), glucocorticoid receptor (GR), and MAPKs (ERK, JNK, and p38).

Results

Total AR were down-regulated post-exercise at T2 in NFOR only. Phospho-AR at ser515 increased in both groups post-exercise at T1; however, ser515 only increased at T2 in NFOR. Phosphorylated ERK, JNK, and p38 increased post-exercise in CON and NFOR at T1 and T2. Post-exercise phospho-p38 was blunted in NFOR at T2 compared to T1. After the training intervention, resting phospho-p38 was higher in NFOR compared to T1. At T2, post-exercise phospho-GR at ser226 was lower compared to T1, and resting levels increased in NFOR.

Conclusion

Steroid receptors are phosphorylated after acute resistance exercise, and in addition to MAPKs, are differentially regulated after stressful training with insufficient recovery.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

1RM:

One-repetition maximum

AR:

Androgen receptor

β2-AR:

Β2 adrenergic receptors

CON:

Control group

ERK:

Extracellular signaling-regulated kinase 1/2

ELISA:

Enzyme-linked immunosorbent assay

IR:

Infrared

JNK:

c-Jun NH2-terminal kinase

GR:

Glucocorticoid receptor

MAPK:

Mitogen-activated protein kinase

NFOR:

Non-functional overreaching

PVDF:

Polyvinylidene difluoride

RE:

Resistance exercise

PMANOVA:

Repeated-measures analysis of variance

RT:

Resistance training

SDS:

Sodium dodecyl sulfate

TBS:

Tris-buffered saline

TBST:

Tris-buffered saline plus Tween 20

TNF-α:

Tumor necrosis factor-α

References

  • Alves Souza RW, Aguiar AF, Vechetti-Júnior IJ, Piedade WP, Rocha Campos GE, Dal-Pai-Silva M (2014) Resistance training with excessive training load and insufficient recovery alters skeletal muscle mass- related protein expression. J Strength Cond Res 28:2338–2345

    Article  Google Scholar 

  • Baechle TR, Earle RW (2008) Resistance training and spotting techniques. In: Baechle TR, Earle RW (eds) Essentials of strength training and conditioning. Human Kinetics, Champaign, pp 326–376

    Google Scholar 

  • Boncompagni S, Arthurton L, Akujuru E, Pearson T, Steverding D, Protasi F, Mutungi G (2015) Membrane glucocorticoid receptors are localized in the extracellular matrix and signal through the MAPK pathway in mammalian skeletal muscle fibers. J Physiol 593:2679–2692

    Article  CAS  Google Scholar 

  • Brook MS, Wilkinson DJ, Mitchell WK, Lund JN, Phillips BE, Szewczyk NJ, Greenhaff PL, Smith K, Atherton PJ (2016) Synchronous deficits in cumulative muscle protein synthesis and ribosomal biogenesis underline age-related anabolic resistance to exercise in humans. J Physiol 594:7399–7417

    Article  CAS  Google Scholar 

  • Cadegiani FA, Kater CE (2017) Hormonal aspects of overtraining syndrome: a systematic review. BMC Sports Sci Med Rehabil 9:14

    Article  Google Scholar 

  • Chen W, Dang T, Blind RD, Wang Z, Cavasotto CN, Hittelman AB, Rogatsky I, Logan SK, Garabedian MJ (2008) Glucocorticoid receptor phosphorylation differentially affects target gene expression. Mol Endocrinol 22:1754–1766

    Article  CAS  Google Scholar 

  • Coffey VG, Zhong Z, Shield A, Canny BJ, Chibalin AV, Zierath JP, Hawley JA (2006) Early signaling responses to divergent exercise stimuli in skeletal muscle from well-trained humans. FASEB J 20:190–192

    Article  CAS  Google Scholar 

  • Coffey VG, Reeder DW, Lancaster GI, Yeo WK, Febbraio MA, Yaspelkis BB III, Hawley JA (2007) Effect of high-frequency resistance exercise on adaptive responsive in skeletal muscle. Med Sci Sports Exerc 39:2135–2144

    Article  Google Scholar 

  • Coutinho AE, Campbell JE, Frediuc S, Riddell MC (2006) Effect of voluntary exercise on peripheral tissue glucocorticoid receptor content and the expression and activity of 11β -HSD1 in the Syrian hamster. J Appl Physiol 100:1483–1488

    Article  CAS  Google Scholar 

  • Creer A, Gallagher P, Slivka D, Jemiolo B, Fink W, Trappe S (2005) Influence of muscle glycogen availability on ERK1/2 and Akt signaling after resistance exercise in human skeletal muscle. J Appl Physiol 99:950–956

    Article  CAS  Google Scholar 

  • Da Rocha AL, Pereira BC, Teixeira GR, Pinto AP, Frantz FG, Elias LLK, Lira FS, Pauli JR, Cintra DE, Ropoelle ER, de Moura LP, Mekary RA, de Freitas EC, da Silva ASR (2017) Treadmill slope modulates inflammation, fiber type composition, androgen and glucocorticoid receptors in skeletal muscle of overtrained mice. Front Immunol 8:1378. https://doi.org/10.3389/fimmu.2017.01378

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Estrada M, Espinosa A, Müller M, Jaimovich E (2003) Testosterone stimulates intracellular calcium release and mitogen-activated protein kinases via a G protein-coupled receptor in skeletal muscle cells. Endocrinology 144:3586–3597

    Article  CAS  Google Scholar 

  • Evans WJ, Phinney SD, Young VR (1982) Suction applied to a muscle biopsy maximizes sample size. Med Sci Sport Exerc 14:101–102

    CAS  Google Scholar 

  • Figueiredo VC, Caldow MK, Massie V, Markworth JF, Cameron-Smith D, Blazevich AJ (2015) Ribosome biogenesis adaptation in resistance training-induced human skeletal muscle hypertrophy. Am J Physiol Endocrinol Metab 309:E72–E83

    Article  CAS  Google Scholar 

  • Fink J, Schoenfeld BJ, Nakazato K (2017) The role of hormones in muscle hypertrophy. Phys Sportsmed 46:129–134

    Article  Google Scholar 

  • Fry AC (2004) The role of resistance exercise intensity on muscle fibre adaptations. Sports Med 34:663–679

    Article  Google Scholar 

  • Fry AC, Kraemer WJ (1997) Resistance exercise overtraining and overreaching: neuroendocrine responses. Sports Med 23:106–129

    Article  CAS  Google Scholar 

  • Fry AC, Kraemer WJ, Stone MJ, Warren BJ, Fleck SJ, Kearney JT, Gordon SE (1994) Endocrine responses to overreaching before and after 1 year of weightlifting. Can J Appl Physiol 19:400–410

    Article  CAS  Google Scholar 

  • Fry AC, Kraemer WJ, Ramsey LT (1998) Pituitary-adrenal-gonadal responses to high-intensity resistance exercise overtraining. J Appl Physiol 85:2352–2359

    Article  CAS  Google Scholar 

  • Fry AC, Schilling BK, Weiss LW, Chiu LZF (2006) β2 Adrenergic receptor downregulation and performance decrements during high-intensity resistance exercise overtraining. J Appl Physiol 101:1664–1672

    Article  CAS  Google Scholar 

  • Fujii N, Boppart MD, Dufresne SD, Crowley PF, Jozsi AC, Sakamoto K, Yu H, Aschenbach WG, Kim S, Rui L, White MF, Hirshman MF, Goodyear LJ (2004) Overexpression or ablation of JNK in skeletal muscle has no effect on glycogen synthase activity. Am J Physiol Cell Physiol 287:C200–C208

    Article  CAS  Google Scholar 

  • Galliher-Beckley AJ, Williams JG, Cidlowski JA (2011) Ligand-independent phosphorylation of the glucocorticoid receptor integrates cellular stress pathways with nuclear receptor signaling. Mol Cell Biol 31:4663–4675

    Article  CAS  Google Scholar 

  • Galpin AJ, Fry AC, Nicoll JX, Moore CA, Schilling BK, Thomason DB (2016) Resting extracellular signal-regulated protein kinase 1/2 expression following a continuum of chronic resistance exercise training paradigms. Res Sports Med 24:298–303

    Article  Google Scholar 

  • Gehlert S, Suhr F, Gutsche K, Willkomm L, Kern J, Jacko D, Knicker A, Schiffer T, Wackerhage H, Bloch W (2015) High force development augments skeletal muscle signalling in resistance exercise modes equalized for time under tension. Pflugers Arch 467:1343–1356

    Article  CAS  Google Scholar 

  • Gonzalez AM, Hoffman JR, Townsend JR, Jajtner AR, Boone CH, Beyer KS, Baker KM, Wells AJ, Mangine GR, Robinson EH, Church DD, Oliveira LP, Fakuda DH, Stout JR (2016) Intramuscular MAPK signaling following high volume and high intensity resistance exercise protocols in trained men. Eur J Appl Physiol 116:1663–1670

    Article  CAS  Google Scholar 

  • Holm S (1979) A simple sequential rejective multiple test procedure. Scand J Statist 6:65–70

    Google Scholar 

  • Hughes DC, Stewart CE, Sculthorpe N, Dugdale HF, Yousefian F, Lewis MP, Sharples AP (2016) Testosterone enables growth and hypertrophy in fusion impaired myoblasts that display myotube atrophy: deciphering the role of androgen and IGF-I receptors. Biogerontology 17:619–639

    Article  CAS  Google Scholar 

  • Itoh M, Adachi M, Yasui H, Takekawa M, Tanaka H, Imai K (2002) Nuclear export of glucocorticoid receptor is enhanced by c-Jun N-terminal kinase-mediated phosphorylation. Mol Endocrinol 16:2382–2392

    Article  CAS  Google Scholar 

  • Karagounis LG, Yaspelkis BB, Reeder DW, Lancaster GI, Hawley JA, Coffey VG (2010) Contraction-induced changes in TNF-α and Akt-mediated signaling are associated with increased myofibrillar protein in rate skeletal muscle. Eur J Appl Physiol 109:839–848

    Article  CAS  Google Scholar 

  • Kim HJ, Lee WJ (2009) Ligand-independent activation of the androgen receptor by insulin-like growth factor-1 and the role of the MAPK pathway in skeletal muscle cells. Mol Cells 28:589–593

    Article  CAS  Google Scholar 

  • Koryakina Y, Ta HQ, Gioeli D (2014) Androgen receptor phosphorylation: biological context and functional consequences. Endocrin Relat Cancer 21:T131–T145

    Article  CAS  Google Scholar 

  • Kraemer WJ, Spiering BA, Volek JS, Ratamess NA, Sharman MJ, Rubin MR, French DN, Silvestre R, Hatfield DL, Van Heest JL, Vingren JL, Judelson DA, Deschenes MR, Maresh CM (2006a) Androgenic responses to resistance exercise: effects of feeding and l-carnitine. Med Sci Sports Exerc 38(7):1288–1296

    Article  CAS  Google Scholar 

  • Kraemer WJ, Ratamess NA, Fry AC, French DN (2006b) Strength training: development and evaluation of methodology. In: Maud PJ, Foster C (eds) Physiological assessment of human fitness. Human Kinetics, Champaign, pp 119–150

    Google Scholar 

  • Kudrna RA, Fry AC, Nicoll JX, Gallagher PM, Prewitt MR (2019) The effect of three different maximal concentric velocity squat protocols on MAPK phosphorylation and endocrine responses. J Strength Cond Res 33(6):1692–1702. https://doi.org/10.1519/jsc.0000000000002411

    Article  PubMed  Google Scholar 

  • Lee JS, Bruce CR, Spurrell BE, Hawley JA (2002) Effect of training on activation of extracellular signal-regulated kinase 1/2 and p38 mitogen-activated protein kinase pathways in rat soleus muscle. Clin Exp Pharmacol Physiol 29:655–660

    Article  CAS  Google Scholar 

  • Lessard SJ, MacDOnald TL, Pathak P, Han MS, Coffey VG, Edge J, Rivas DA, Hirshman MF, Davis RJ, Goodyear LJ (2018) JNK regulates muscle remodeling via myostatin/SMAD inhibition. Nat Commun 9(1):3030

    Article  Google Scholar 

  • Li YP, Chen Y, John J, Moylan J, Jin B, Mann DL, Reid MB (2005) TNF- α acts via p38 MAPK to stimulate expression of the ubiquitin ligase atrogin1/MAFbx in skeletal muscle. FASEB J 19:362–370

    Article  CAS  Google Scholar 

  • Macaluso F, Brooks NE, van de Vyer M, Niesler CU, Myburgh KH (2012) Satellite cell count, VO2max, and p38 MAPK in active to moderately active young men. Scand J Med Sci Sports 22:e38–e44

    Article  CAS  Google Scholar 

  • Mangine GT, Hoffman JR, Gonzalez AM, Townsend JR, Wells AJ, Jajtner AR, Beyer KS, Boone CH, Wang R, Miramonti AA, LaMonica MB, Fukuda DH, Witta EL, Ratamess NA, Stout JR (2017) Exercise-induced hormone elevations are related to muscle growth. J Strength Cond Res 31:45–53

    Article  Google Scholar 

  • Martineau LC, Gardiner PF (2001) Insight into skeletal muscle mechanotransduction: MAPK activation is quantitatively related to tension. J Appl Physiol 91:693–702

    Article  CAS  Google Scholar 

  • Meeusen R, Duclos M, Foster C, Fry A, Nieman D, Raglin J, Rietjens G, Steinacker J, Urhausen A, European College of Sport Science; American College of Sports Medicine (2013) Prevention, diagnosis, and treatment of the overtraining syndrome: joint consensus statement of the European College of Sport Science and the American College of Sports Medicine. Med Sci Sports Exerc 45:186–205

    Article  Google Scholar 

  • Mitchell CJ, Churchward-Venne TA, Bellamy L, Parise G, Baker SK, Phillips SM (2013) Muscular and systemic correlates of resistance training-induced muscle hypertrophy. PLoS One 8:e78636

    Article  CAS  Google Scholar 

  • Mobley CB, Haun CT, Robertson PA, Mumford PW, Kepheart WC, Romero MA, Vann CG, Young KC, Beck DT, Martin JS, Lockwood CM, Roberts MD (2018) Biomarkers associated with low, moderate, and high vastus lateralis muscle hypertrophy following 12 weeks of resistance training. PLoS One 13(4):e0195203. https://doi.org/10.1371/journal.pone.0195203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nicoll JX, Fry AC, Galpin AJ, Sterczala AJ, Thomason DB, Moore CA, Weiss LW, Chiu LZF (2016) Changes in resting mitogen-activated protein kinases following resistance exercise overreaching and overtraining. Eur J Appl Physiol 116:2401–2413

    Article  CAS  Google Scholar 

  • Nicoll JX, Fry AC, Galpin AJ, Thomason DB, Moore CA (2017) Resting MAPK expression in chronically trained endurance runners. Eur J Sport Sci 17(9):1194–1202

    Article  Google Scholar 

  • Nicoll JX, Fry AC, Mosier EM (2019) Sex-based differences in resting MAPK, androgen, and glucocorticoid receptor phosphorylation in human skeletal muscle. Steroids 141:23–29

    Article  CAS  Google Scholar 

  • Pereira BC, Pauli JR, De Souza CT, Ropelle ER, Cintra DE, Freitas EC, da Silva AS (2014) Eccentric exercise leads to performance decrease in insulin signaling impairment. Med Sci Sports Exerc 46:686–694

    Article  Google Scholar 

  • Petrella JK, Kim JS, Meyhew DL, Cross JM, Bamman MM (2008) Potent myofiber hypertrophy during resistance training in humans is associated with satellite cell-mediated myonuclear addition: a cluster analysis. J Appl Physiol 104:1736–1742

    Article  Google Scholar 

  • Ponguta LA, Gregory CW, French FS, Wilson EM (2008) Site-specific androgen receptor serine phosphorylation linked to epidermal growth factor-dependent growth of castration-recurrent prostate cancer. J Biol Chem 283:20989–21001

    Article  CAS  Google Scholar 

  • Raastad T, Glomsheller T, Bjøro T, Hallén J (2001) Changes in human skeletal muscle contractility and hormone status during 2 weeks of heavy strength training. Eur J Appl Physiol 84:54–63

    Article  CAS  Google Scholar 

  • Raastad T, Glomsheller T, Bjøro T, Hallén J (2003) Recovery of skeletal muscle contractility and hormonal responses to strength exercise after two weeks of high-volume strength training. Scand J Med Sci Sports 13:159–168

    Article  CAS  Google Scholar 

  • Ratamess NA, Kraemer WJ, Volek JS, Maresh CM, VanHeest JL, Sharman MJ, Rubin MR, French DN, Vescovi JD, Silvestre R, Hatfield DL, Fleck SJ, Deschenes MR (2005) Androgen receptor content following heavy resistance exercise in men. J Steroid Biochem Mol Biol 93:35–42

    Article  CAS  Google Scholar 

  • Spiering BA, Kraemer WJ, Vingren JL, Ratamess NA, Anderson JM, Armstrong LE, Nindl BC, Volek JS, Häkkinen K, Maresh CM (2009) Elevated endogenous testosterone concentrations potentiate muscle androgen receptor responses to resistance exercise. J Steroid Biochem and Mol Biol 114:195–199

    Article  CAS  Google Scholar 

  • Spillane M, Schwarz N, Willoughby DS (2015) Upper-body resistance exercise vastus lateralis androgen receptor-DNA binding and canonical Wnt/B-catenin signaling compared to lower-body resistance exercise in resistance-trained men without an increase in serum testosterone. Steroids 98:63–71

    Article  CAS  Google Scholar 

  • Staron RS (1991) Correlation between myofibrillar ATPase activity and myosin heavy chain composition in single human muscle fibers. Histochemistry 96:21–24

    Article  CAS  Google Scholar 

  • Sterczala AJ, Fry AC, Chiu LZF, Schilling BK, Weiss LW, Nicoll JX (2017) β2-adrenergic receptor maladaptations to high power resistance exercise overreaching. Hum Physiol (Fiziologiya Cheloveka) 43(4):446–454. https://doi.org/10.1134/S0362119717040144

    Article  CAS  Google Scholar 

  • Takegaki J, Ogaswara R, Tamura Y, Takagi R, Arihara Y, Tsutaki A, Nakazato K, Ishii N (2017) Repeated bouts of resistance exercise with short recovery periods activates mTOR signaling, not protein synthesis in mouse skeletal muscle. Physiol Rep 5:e13515

    Article  Google Scholar 

  • Vichaiwong K, Henriksen EJ, Toskulkao C, Prasannarong M, Bupha-Intr T, Saengsirisuwan V (2009) Attenuation of oxidant-induced muscle insulin resistance and p38 MAPK by exercise training. Free Redic Biol Med 47:593–599

    Article  CAS  Google Scholar 

  • Vingren JL, Kraemer WJ, Hatfield DL, Volek JS, Ratamess NA, Anderson JM, Häkkinen K, Ahtianen J, Fragala MS, Thomas GA, Ho JY, Maresh CM (2009) Effect of resistance exercise on muscle steroid receptor content in strength-trained men and women. Steroids 74(13–14):1033–1039

    Article  CAS  Google Scholar 

  • Wilder JM, Heng SJ, McCall P, Adams CE, Tannahill C, Fyffe G, Seywright M, Horgan PG, Leung HY, Underwood MA, Edwards J (2013) Androgen receptor phosphorylation at serine 515 by Cdk1 predicts biochemical relapse in prostate cancer patients. Br J Cancer 108:139–148

    Article  Google Scholar 

  • Willoughby DS, Taylor L (2004) Effects of sequential bouts of resistance exercise on androgen receptor expression. Med Sci Sports Exerc 36:1499–1506

    Article  CAS  Google Scholar 

  • Willoughby DS, Taylor M, Taylor L (2003) Glucocorticoid receptor and ubiquitin expression after repeated eccentric exercise. Med Sci Sports Exerc 35:2023–2031

    Article  CAS  Google Scholar 

  • Wirth K, Hartmann H, Sander A, Mickel C, Szilvas E, Keiner M (2016) The impact of back squat and leg-press exercises on maximal strength and speed-strength parameters. J Strength Cond Res 30:1205–1212

    Article  Google Scholar 

  • Yeh S, Lin HK, Kang HY, Thin TH, Lin MF, Chang C (1999) From HER2/Neu signal cascade to androgen receptor and its coactivators: a novel pathway by indication of androgen target genes through MAPK kinase in prostate cancer cells. Proc Natl Acad Sci USA 96:5458–5463

    Article  CAS  Google Scholar 

  • Yu M, Blomstrand E, Chibalin AV, Wallberg-Henriksson H, Zierath JR, Krook A (2001) Exercise-associated differences in an array of proteins involved in signal transduction and glucose transport. J Appl Physiol 90:29–34

    Article  CAS  Google Scholar 

  • Zeng F, Zhao H, Liao J (2017) Androgen interacts with exercise through the mTOR pathway to induce skeletal muscle hypertrophy. Biol Sport 34:313–321

    Article  Google Scholar 

Download references

Funding

This study was funded in part through a research grant from the National Strength and Conditioning Association and University of Kansas Graduate Research Grant

Author information

Authors and Affiliations

  1. Department of Kinesiology, California State University, Northridge, 18111 Nordhoff St, Northridge, CA, 91330-8287, USA

    Justin X. Nicoll

  2. Department of Health, Sport, and Exercise Sciences, University of Kansas, Lawrence, KS, 66045, USA

    Andrew C. Fry & Stephanie A. Sontag

  3. School of Health Science and Wellness, Northwest Missouri State University, Maryville, MO, 64468, USA

    Eric M. Mosier

  4. Biomedical Sciences, University of Kansas Medical Center, Kansas City, KS, 66160, USA

    Luke A. Olsen

Authors
  1. Justin X. Nicoll
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrew C. Fry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric M. Mosier
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luke A. Olsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stephanie A. Sontag
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Study design and conceptualization: JXN and ACF. Data collection and analysis JXN, ACF, EMM, LO, and SAS. Writing: JXN. Revision: JXN, ACF, EMM, LAO, and SAS. Funding: JXN and ACF.

Corresponding author

Correspondence to Justin X. Nicoll.

Ethics declarations

Conflict of interest

The authors report no conflict of interest.

Additional information

Communicated by William J. Kraemer.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nicoll, J.X., Fry, A.C., Mosier, E.M. et al. MAPK, androgen, and glucocorticoid receptor phosphorylation following high-frequency resistance exercise non-functional overreaching. Eur J Appl Physiol 119, 2237–2253 (2019). https://doi.org/10.1007/s00421-019-04200-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00421-019-04200-y

Keywords

Search

Navigation