Modulation of autophagy and ubiquitin-proteasome pathways during ultra-endurance running.

In this study, the coordinated activation of ubiquitin-proteasome pathway (UPP), autophagy-lysosomal pathway (ALP), and mitochondrial remodeling including mitophagy was assessed by measuring protein markers during ultra-endurance running exercise in human skeletal muscle. Eleven male, experienced ultra-endurance athletes ran for 24 h on a treadmill. Muscle biopsy samples were taken from the vastus lateralis muscle 2 h before starting and immediately after finishing exercise. Athletes ran 149.8 ± 16.3 km with an effective running time of 18 h 42 min ( ± 41 min). The phosphorylation state of Akt (-74 ± 5%; P < 0.001), FOXO3a (-49 ± 9%; P < 0.001), mTOR Ser2448 (-32 ± 14%; P = 0.028), and 4E-BP1 (-34 ± 7%; P < 0.001) was decreased, whereas AMPK phosphorylation state increased by 247 ± 170% (P = 0.042). Proteasome β2 subunit activity increased by 95 ± 44% (P = 0.028), whereas the activities associated with the β1 and β5 subunits remained unchanged. MuRF1 protein level increased by 55 ± 26% (P = 0.034), whereas MAFbx protein and ubiquitin-conjugated protein levels did not change. LC3bII increased by 554 ± 256% (P = 0.005), and the form of ATG12 conjugated to ATG5 increased by 36 ± 17% (P = 0.042). The mitochondrial fission marker phospho-DRP1 increased by 110 ± 47% (P = 0.003), whereas the fusion marker Mfn1 and the mitophagy markers Parkin and PINK1 remained unchanged. These results fit well with a coordinated regulation of ALP and UPP triggered by FOXO3 and AMPK during ultra-endurance exercise.

[1]  M. Francaux,et al.  Autophagy-related and autophagy-regulatory genes are induced in human muscle after ultraendurance exercise , 2012, European Journal of Applied Physiology.

[2]  K. Wertz,et al.  Mitochondrial dynamic remodeling in strenuous exercise-induced muscle and mitochondrial dysfunction: regulatory effects of hydroxytyrosol. , 2011, Free radical biology & medicine.

[3]  O. Senn,et al.  Do ultra-runners in a 24-h run really dehydrate? , 2011, Irish journal of medical science.

[4]  G. Millet,et al.  Physiological and biological factors associated with a 24 h treadmill ultra‐marathon performance , 2011, Scandinavian journal of medicine & science in sports.

[5]  Alexander Doyle,et al.  Toll‐like receptor 4 mediates lipopolysaccharide‐induced muscle catabolism via coordinate activation of ubiquitin‐proteasome and autophagy‐lysosome pathways , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[6]  T. Trappe,et al.  Muscle protein synthesis and gene expression during recovery from aerobic exercise in the fasted and fed states. , 2010, American journal of physiology. Regulatory, integrative and comparative physiology.

[7]  Michael J. Short,et al.  Early time course of Akt phosphorylation after endurance and resistance exercise. , 2010, Medicine and science in sports and exercise.

[8]  C. Mattsson,et al.  Energy turnover during 24 hours and 6 days of adventure racing , 2010, Journal of sports sciences.

[9]  S. Barth,et al.  Autophagy: assays and artifacts , 2010, The Journal of pathology.

[10]  Luca Scorrano,et al.  Mitochondrial fission and remodelling contributes to muscle atrophy , 2010, The EMBO journal.

[11]  G. Millet,et al.  Central and peripheral contributions to neuromuscular fatigue induced by a 24-h treadmill run. , 2010, Journal of applied physiology.

[12]  Chang Hwa Jung,et al.  mTOR regulation of autophagy , 2010, FEBS letters.

[13]  C. Mattsson,et al.  Ultraendurance exercise increases the production of reactive oxygen species in isolated mitochondria from human skeletal muscle. , 2010, Journal of applied physiology.

[14]  L. Ji,et al.  Response of mitochondrial fusion and fission protein gene expression to exercise in rat skeletal muscle. , 2010, Biochimica et biophysica acta.

[15]  Atsushi Tanaka,et al.  PINK1 Is Selectively Stabilized on Impaired Mitochondria to Activate Parkin , 2010, PLoS biology.

[16]  D. Fingar,et al.  mTOR Ser-2481 Autophosphorylation Monitors mTORC-specific Catalytic Activity and Clarifies Rapamycin Mechanism of Action* , 2009, The Journal of Biological Chemistry.

[17]  D. Metzger,et al.  Autophagy is required to maintain muscle mass. , 2009, Cell metabolism.

[18]  Ji Zhang,et al.  Role of BNIP3 and NIX in cell death, autophagy, and mitophagy , 2009, Cell Death and Differentiation.

[19]  E. Richter,et al.  AMP‐activated protein kinase in contraction regulation of skeletal muscle metabolism: necessary and/or sufficient? , 2009, Acta physiologica.

[20]  Eeva-Liisa Eskelinen,et al.  Autophagy: a lysosomal degradation pathway with a central role in health and disease. , 2009, Biochimica et biophysica acta.

[21]  T. Trappe,et al.  Protein synthesis and the expression of growth-related genes are altered by running in human vastus lateralis and soleus muscles. , 2009, American journal of physiology. Regulatory, integrative and comparative physiology.

[22]  YongSung Kim,et al.  PINK1 controls mitochondrial localization of Parkin through direct phosphorylation. , 2008, Biochemical and biophysical research communications.

[23]  V. Hill,et al.  Suppression of autophagy in skeletal muscle uncovers the accumulation of ubiquitinated proteins and their potential role in muscle damage in Pompe disease. , 2008, Human molecular genetics.

[24]  J. Zierath,et al.  Divergent cell signaling after short-term intensified endurance training in human skeletal muscle. , 2008, American journal of physiology. Endocrinology and metabolism.

[25]  D. Theisen,et al.  Decrease in Akt/PKB signalling in human skeletal muscle by resistance exercise , 2008, European Journal of Applied Physiology.

[26]  Marco Sandri,et al.  Signaling in muscle atrophy and hypertrophy. , 2008, Physiology.

[27]  C. Mammucari,et al.  Downstream of Akt: FoxO3 and mTOR in the regulation of autophagy in skeletal muscle , 2008, Autophagy.

[28]  P. Hasselgren,et al.  The NF-κB Inhibitor Curcumin Blocks Sepsis-Induced Muscle Proteolysis , 2008, Mediators of inflammation.

[29]  A. Young,et al.  Increased protein maintains nitrogen balance during exercise-induced energy deficit. , 2008, Medicine and science in sports and exercise.

[30]  A. Goldberg,et al.  Coordinate activation of autophagy and the proteasome pathway by FoxO transcription factor , 2008, Autophagy.

[31]  Min Wu,et al.  Fission and selective fusion govern mitochondrial segregation and elimination by autophagy , 2008, The EMBO journal.

[32]  A. Goldberg,et al.  FoxO3 controls autophagy in skeletal muscle in vivo. , 2007, Cell metabolism.

[33]  A. Goldberg,et al.  FoxO3 coordinately activates protein degradation by the autophagic/lysosomal and proteasomal pathways in atrophying muscle cells. , 2007, Cell metabolism.

[34]  S. Trappe,et al.  Time course of proteolytic, cytokine, and myostatin gene expression after acute exercise in human skeletal muscle. , 2007, Journal of applied physiology.

[35]  S. Gygi,et al.  The Energy Sensor AMP-activated Protein Kinase Directly Regulates the Mammalian FOXO3 Transcription Factor* , 2007, Journal of Biological Chemistry.

[36]  S. Strack,et al.  Reversible phosphorylation of Drp1 by cyclic AMP‐dependent protein kinase and calcineurin regulates mitochondrial fission and cell death , 2007, EMBO reports.

[37]  H. Mascher,et al.  Changes in signalling pathways regulating protein synthesis in human muscle in the recovery period after endurance exercise , 2007, Acta physiologica.

[38]  K. Nakashima,et al.  AMPK Activation Stimulates Myofibrillar Protein Degradation and Expression of Atrophy-Related Ubiquitin Ligases by Increasing FOXO Transcription Factors in C2C12 Myotubes , 2007, Bioscience, biotechnology, and biochemistry.

[39]  Y. Ohsumi,et al.  Atg8, a Ubiquitin-like Protein Required for Autophagosome Formation, Mediates Membrane Tethering and Hemifusion , 2007, Cell.

[40]  C. Bauvy,et al.  AMP-activated Protein Kinase and the Regulation of Autophagic Proteolysis* , 2006, Journal of Biological Chemistry.

[41]  E. Taylor,et al.  Role of AMP-activated protein kinase in the molecular adaptation to endurance exercise. , 2006, Medicine and science in sports and exercise.

[42]  D. Chan.  Mitochondrial fusion and fission in mammals. , 2006, Annual review of cell and developmental biology.

[43]  Anthony Shield,et al.  Early signaling responses to divergent exercise stimuli in skeletal muscle from well‐trained humans , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[44]  I. Rieu,et al.  A leucine‐supplemented diet restores the defective postprandial inhibition of proteasome‐dependent proteolysis in aged rat skeletal muscle , 2005, The Journal of physiology.

[45]  M. Reid.  Response of the ubiquitin-proteasome pathway to changes in muscle activity. , 2005, American journal of physiology. Regulatory, integrative and comparative physiology.

[46]  M. Tarnopolsky.  Protein requirements for endurance athletes. , 2004, Nutrition.

[47]  G. Yancopoulos,et al.  The IGF-1/PI3K/Akt pathway prevents expression of muscle atrophy-induced ubiquitin ligases by inhibiting FOXO transcription factors. , 2004, Molecular cell.

[48]  A. Goldberg,et al.  Multiple types of skeletal muscle atrophy involve a common program of changes in gene expression , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[49]  M. Matsui,et al.  In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. , 2003, Molecular biology of the cell.

[50]  D. Attaix,et al.  Class III phosphoinositide 3-kinase--Beclin1 complex mediates the amino acid-dependent regulation of autophagy in C2C12 myotubes. , 2003, The Biochemical journal.

[51]  B. Kemp,et al.  AMPK β Subunit Targets Metabolic Stress Sensing to Glycogen , 2003, Current Biology.

[52]  R. Candau,et al.  Effects of concurrent endurance and strength training on running economy and .VO(2) kinetics. , 2002, Medicine and science in sports and exercise.

[53]  D. Bolster,et al.  AMP-activated Protein Kinase Suppresses Protein Synthesis in Rat Skeletal Muscle through Down-regulated Mammalian Target of Rapamycin (mTOR) Signaling* , 2002, The Journal of Biological Chemistry.

[54]  S. Bodine,et al.  Control of Ser2448 Phosphorylation in the Mammalian Target of Rapamycin by Insulin and Skeletal Muscle Load* , 2002, The Journal of Biological Chemistry.

[55]  K Suzuki,et al.  The pre‐autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation , 2001, The EMBO journal.

[56]  D J Glass,et al.  Identification of Ubiquitin Ligases Required for Skeletal Muscle Atrophy , 2001, Science.

[57]  D. Alessi,et al.  Mammalian target of rapamycin is a direct target for protein kinase B: identification of a convergence point for opposing effects of insulin and amino-acid deficiency on protein translation. , 1999, The Biochemical journal.

[58]  M. Birnbaum,et al.  Insulin, but Not Contraction, Activates Akt/PKB in Isolated Rat Skeletal Muscle* , 1998, The Journal of Biological Chemistry.

[59]  A. Goldberg,et al.  Importance of the ATP-Ubiquitin-Proteasome Pathway in the Degradation of Soluble and Myofibrillar Proteins in Rabbit Muscle Extracts* , 1996, The Journal of Biological Chemistry.

[60]  H. Kuipers,et al.  Exercise-Induced Muscle Damage , 1994, International journal of sports medicine.

[61]  K. Henriksson.  “Semi‐open” muscle biopsy technique A SIMPLE OUTPATIENT PROCEDURE , 1979, Acta neurologica Scandinavica.

[62]  A. Sims,et al.  Discriminators between neurotics who die and neurotics who live , 1979, Acta psychiatrica Scandinavica.

[63]  W. Malaisse,et al.  Effects of epinephrine, stress, and exercise on insulin secretion by the rat. , 1968, The American journal of physiology.

[64]  Hyo Jeong Kim,et al.  ER-Stress Markers and Ubiquitin-Proteasome Pathway Activity in Response to 200-Km Run. , 2011 .

[65]  DAVID J. Smith,et al.  Training principles and issues for ultra-endurance athletes , 2005, Current sports medicine reports.

[66]  D. Leroith,et al.  Skeletal muscle. , 2005, Advances in experimental medicine and biology.

[67]  B. Kemp,et al.  AMPK beta subunit targets metabolic stress sensing to glycogen. , 2003, Current biology : CB.