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Purinergic receptors expressed in human skeletal muscle fibres

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Abstract

Purinergic receptors are present in most tissues and thought to be involved in various signalling pathways, including neural signalling, cell metabolism and local regulation of the microcirculation in skeletal muscles. The present study aims to determine the distribution and intracellular content of purinergic receptors in skeletal muscle fibres in patients with type 2 diabetes and age-matched controls. Muscle biopsies from vastus lateralis were obtained from six type 2 diabetic patients and seven age-matched controls. Purinergic receptors were analysed using light and confocal microscopy in immunolabelled transverse sections of muscle biopsies. The receptors P2Y4, P2Y11 and likely P2X1 were present intracellularly or in the plasma membrane of muscle fibres and were thus selected for further detailed morphological analysis. P2X1 receptors were expressed in intracellular vesicles and sarcolemma. P2Y4 receptors were present in sarcolemma. P2Y11 receptors were abundantly and diffusely expressed intracellularly and were more explicitly expressed in type I than in type II fibres, whereas P2X1 and P2Y4 showed no fibre-type specificity. Both diabetic patients and healthy controls showed similar distribution of receptors. The current study demonstrates that purinergic receptors are located intracellularly in human skeletal muscle fibres. The similar cellular localization of receptors in healthy and diabetic subjects suggests that diabetes is not associated with an altered distribution of purinergic receptors in skeletal muscle fibres. We speculate that the intracellular localization of purinergic receptors may reflect a role in regulation of muscle metabolism; further studies are nevertheless needed to determine the function of the purinergic system in skeletal muscle cells.

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References

  1. Ralevic V, Burnstock G (1998) Receptors for purines and pyrimidines. Pharmacol Rev 50:413–492

    PubMed  CAS  Google Scholar 

  2. Erlinge D, Burnstock G (2008) P2 receptors in cardiovascular regulation and disease. Purinergic Signal 4:1–20

    Article  PubMed  CAS  Google Scholar 

  3. Burnstock G (2004) Introduction: P2 receptors. Curr Top Med Chem 4:793–803

    Article  PubMed  CAS  Google Scholar 

  4. Abbracchio M, Burnstock G, Boeynaems J, Barnard E, Boyer J, Kennedy C, Knight G, Fumagalli M, Gachet C, Jacobson K, Weisman G (2006) International Union of Pharmacology LVIII: update on the P2Y G protein-coupled nucleotide receptors: from molecular mechanisms and pathophysiology to therapy. Pharmacol Rev 58:281–341

    Article  PubMed  CAS  Google Scholar 

  5. Burnstock G (2007) Purine and pyrimidine receptors. Cell Moll Life Sci 64:1471–1483

    Article  CAS  Google Scholar 

  6. Wang L, Karlsson L, Moses S, Hultgårdh-Nilsson A, Andersson M, Borna C, Gudbjartsson T, Jern S, Erlinge D (2002) P2 receptor expression profiles in human vascular smooth muscle and endothelial cells. J Cardiovasc Pharmacol 40:841–853

    Article  PubMed  CAS  Google Scholar 

  7. Thaning P, Bune LT, Hellsten Y, Pilegaard H, Saltin B, Rosenmeier JB (2010) Attenuated purinergic receptor function in patients with type 2 diabetes. Diabetes 59:182–189

    Article  PubMed  CAS  Google Scholar 

  8. Mortensen SP, Gonzalez-Alonso J, Bune LT, Saltin B, Pilegaard H, Hellsten Y (2009) ATP-induced vasodilation and purinergic receptors in the human leg: roles of nitric oxide, prostaglandins, and adenosine. Am J Physiol Regul Integr Comp Physiol 296:R1140–R1148

    Article  PubMed  CAS  Google Scholar 

  9. Mortensen SP, Gonzalez-Alonso J, Nielsen J-J, Saltin B, Hellsten Y (2009) Muscle interstitial ATP and norepinephrine concentrations in the human leg during exercise and ATP infusion. J Appl Physiol 107:1757–1762

    Article  PubMed  CAS  Google Scholar 

  10. Kim MS, Lee J, Ha J, Kim SS, Kong Y, Cho YH, Baik HH, Kang I (2002) ATP stimulates glucose transport through activation of P2 purinergic receptors in C2C12 skeletal muscle cells. Arch Bioch Biophys 401:205–214

    Article  CAS  Google Scholar 

  11. American, Diabetes, Association (2004) Diagnosis and classification of diabetes mellitus. Diabetes Care 27:s5–s10

    Article  Google Scholar 

  12. Bergström J (1975) Percutaneous needle biopsy of skeletal muscle in physiological and clinical research. Scand J Clin Lab Invest 35:609–616

    Article  PubMed  Google Scholar 

  13. Vial C, Evans RJ (2000) P2X receptor expression in mouse urinary bladder and the requirement of P2X1 receptors for functional P2X receptor responses in the mouse urinary bladder smooth muscle. Br J Pharmacol 131:1489–1495

    Article  PubMed  CAS  Google Scholar 

  14. Lewis CJ, Gitterman DP, Schlüter H, Evans RJ (2000) Effects of diadenosine polyphosphates (ApnAs) and adenosine polyphospho guanosines (ApnGs) on rat mesenteric artery P2X receptor ion channels. Br J Pharmacol 129:124–130

    Article  PubMed  CAS  Google Scholar 

  15. Lewis CJ, Evans RJ (2000) Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique, physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels. Br J Pharmacol 131:1659–1666

    Article  PubMed  CAS  Google Scholar 

  16. Kim M, Spelta V, Sim J, North RA, Surprenant A (2001) Differential assembly of rat purinergic P2X7 receptor in immune cells of the brain and periphery. J Biol Chem 276:23262–23267

    Article  PubMed  CAS  Google Scholar 

  17. Gitterman DP, Evans RJ (2000) Properties of P2X and P2Y receptors are dependent on artery diameter in the rat mesenteric bed. Br J Pharmacol 131:1561–1568

    Article  PubMed  CAS  Google Scholar 

  18. Gifford SM, Yi F-X, Bird IM (2006) Pregnancy-enhanced Ca2+ responses to ATP in uterine artery endothelial cells is due to greater capacitative Ca2+ entry rather than altered receptor coupling. J Endocrinol 190:373–384

    Article  PubMed  CAS  Google Scholar 

  19. Ennion S, Hagan S, Evans RJ (2000) The role of positively charged amino acids in ATP recognition by human p2x1 receptors. J Biol Chem 275:29361–29367

    Article  PubMed  CAS  Google Scholar 

  20. Malin S, Molliver D (2010) Gi- and Gq-coupled ADP (P2Y) receptors act in opposition to modulate nociceptive signaling and inflammatory pain behavior. Mol Pain 6:21

    Article  PubMed  Google Scholar 

  21. Ecke D, Hanck T, Tulapurkar ME, Schäfer R, Kassack M, Stricker R, Reiser G (2008) Hetero-oligomerization of the P2Y11 receptor with the P2Y1 receptor controls the internalization and ligand selectivity of the P2Y11 receptor. Biochem J 409:107–116

    Article  PubMed  CAS  Google Scholar 

  22. Lin JHC, Takano T, Arcuino G, Wang X, Hu F, Darzynkiewicz Z, Nunes M, Goldman SA, Nedergaard M (2007) Purinergic signaling regulates neural progenitor cell expansion and neurogenesis. Dev Biol 302:356–366

    Article  PubMed  CAS  Google Scholar 

  23. Knight MM, McGlashan SR, Garcia M, Jensen CG, Poole CA (2009) Articular chondrocytes express connexin 43 hemichannels and P2 receptors—a putative mechanoreceptor complex involving the primary cilium? J Anat 214:275–283

    Article  PubMed  CAS  Google Scholar 

  24. Fischer W, Nörenberg W, Franke H, Schaefer M, Illes P (2009) Increase of intracellular Ca2+ by P2Y but not P2X receptors in cultured cortical multipolar neurons of the rat. J Comp Neurol 516:343–359

    Article  PubMed  CAS  Google Scholar 

  25. Yu J, Sheung N, Soliman EM, Spirli C, Dranoff JA (2009) Transcriptional regulation of IL-6 in bile duct epithelia by extracellular ATP. Am J Physiol Gastrointest Liver Physiol 296:G563–G571

    Article  PubMed  CAS  Google Scholar 

  26. Vaughan KR, Stokes L, Prince LR, Marriott HM, Meis S, Kassack MU, Bingle CD, Sabroe I, Surprenant A, Whyte MKB (2007) Inhibition of neutrophil apoptosis by ATP is mediated by the P2Y11 receptor. J Immunol 179:8544–8553

    PubMed  CAS  Google Scholar 

  27. Lugo-Garcia L, Nadal B, Gomis R, Petit P, Gross R, Lajoix AD (2008) Human pancreatic islets express the purinergic P2Y11 and P2Y12 receptors. Horm Metab Res 40:827–830

    Article  PubMed  CAS  Google Scholar 

  28. Klein C, Grahnert A, Abdelrahman A, Müller CE, Hauschildt S (2009) Extracellular NAD+ induces a rise in [Ca2+]i in activated human monocytes via engagement of P2Y1 and P2Y11 receptors. Cell Calcium 46:263–272

    Article  PubMed  CAS  Google Scholar 

  29. Gulbransen BD, Sharkey KA (2009) Purinergic neuron-to-glia signaling in the enteric nervous system. Gastroenterology 136:1349–1358

    Article  PubMed  CAS  Google Scholar 

  30. Brandenburg L-O, Jansen S, Wruck CJ, Lucius R, Pufe T (2010) Antimicrobial peptide rCRAMP induced glial cell activation through P2Y receptor signalling pathways. Mol Immunol 47:1905–1913

    Article  PubMed  CAS  Google Scholar 

  31. Shanker G, Kontos J, Eckman D, Wesley-Farrington D, Sane D (2006) Nicotine upregulates the expression of P2Y12 on vascular cells and megakaryoblasts. J Thromb Thrombolysis 22:213–220

    Article  PubMed  CAS  Google Scholar 

  32. Ploug T, van Deurs B, Ai H, Cushman SW, Ralston E (1998) Analysis of GLUT4 distribution in whole skeletal muscle fibers: identification of distinct storage compartments that are recruited by insulin and muscle contractions. J Cell Biol 142:1429–1446

    Article  PubMed  CAS  Google Scholar 

  33. von Zglinicki T, Brunk UT (1993) Intracellular interactions under oxidative stress and aging: a hypothesis. Z Gerontol 26:215–220

    Google Scholar 

  34. Sohal RS, Wolfe LS (1986) Chapter 11 lipofuscin: characteristics and significance. Prog Brain Res 70:171–183

    Article  PubMed  CAS  Google Scholar 

  35. Sohal RS, Brunk UT (1989) Lipofuscin as an indicator of oxidative stress and aging. Adv Exp Med Biol 266:17–26

    PubMed  CAS  Google Scholar 

  36. Kiefer CR, McKenney JB, Trainor JF, Lambrecht RW, Bonkovsky HL, Lifshitz LM, Valeri CR, Snyder LM (1998) Porphyrin loading of lipofuscin granules in inflamed striated muscle. Am J Pathol 153:703–708

    Article  PubMed  CAS  Google Scholar 

  37. Ryder J, Chibalin A, Zierath J (2001) Intracellular mechanisms underlying increases in glucose uptake in response to insulin or exercise in skeletal muscle. Acta Physiol Scand 171:249–257

    Article  PubMed  CAS  Google Scholar 

  38. Hellsten Y, Maclean D, Radegran G, Saltin B, Bangsbo J (1998) Adenosine concentrations in the interstitium of resting and contracting human skeletal muscle. Circulation 98:6–8

    PubMed  CAS  Google Scholar 

  39. Durham WJ, Yeckel CW, Miller SL, Gore DC, Wolfe RR (2003) Exogenous nitric oxide increases basal leg glucose uptake in humans. Metabolism 52:662–665

    Article  PubMed  CAS  Google Scholar 

  40. Bradley SJ, Kingwell BA, McConell GK (1999) Nitric oxide synthase inhibition reduces leg glucose uptake but not blood flow during dynamic exercise in humans. Diabetes 48:1815–1821

    Article  PubMed  CAS  Google Scholar 

  41. Kingwell BA, Formosa M, Muhlmann M, Bradley SJ, McConell GK (2002) Nitric oxide synthase inhibition reduces glucose uptake during exercise in individuals with Type 2 diabetes more than in control subjects. Diabetes 51:2572–2580

    Article  PubMed  CAS  Google Scholar 

  42. Ohnishi T, Matsumura S, Ito S (2009) Translocation of neuronal nitric oxide synthase to the plasma membrane by ATP is mediated by P2X and P2Y receptors. Mol Pain 5:40

    Article  PubMed  Google Scholar 

  43. Merry TL, McConell GK (2009) Skeletal muscle glucose uptake during exercise: a focus on reactive oxygen species and nitric oxide signaling. IUBMB Life 61:479–484

    Article  PubMed  CAS  Google Scholar 

  44. Frandsen U, Lopez-Figueroa M, Hellsten Y (1996) Localization of nitric oxide synthase in human skeletal muscle. Biochem Biophys Res Commun 227:88–93

    Article  PubMed  CAS  Google Scholar 

  45. Balogh J, Wihlborg A-K, Isackson H, Joshi BV, Jacobson KA, Arner A, Erlinge D (2005) Phospholipase C and cAMP-dependent positive inotropic effects of ATP in mouse cardiomyocytes via P2Y11-like receptors. J Mol Cell Cardiol 39:223–230

    Article  PubMed  CAS  Google Scholar 

  46. Hou M, Malmsjö M, Möller S, Pantev E, Bergdahl A, Zhao X-H, Sun X-Y, Hedner T, Edvinsson L, Erlinge D (1999) Increase in cardiac P2X1- and P2Y2-receptor mRNA levels in congestive heart failure. Life Sci 65:1195–1206

    Article  PubMed  CAS  Google Scholar 

  47. Dutton JL, Poronnik P, Li GH, Holding CA, Worthington RA, Vandenberg RJ, Cook DI, Barden JA, Bennett MR (2000) P2X1 receptor membrane redistribution and down-regulation visualized by using receptor-coupled green fluorescent protein chimeras. Neuropharmacology 39:2054–2066

    Article  PubMed  CAS  Google Scholar 

  48. Malmsjö M, Edvinsson L, Erlinge D (1998) P2U-receptor mediated endothelium-dependent but nitric oxide-independent vascular relaxation. Br J Pharmacol 123:719–729

    Article  PubMed  Google Scholar 

  49. Dorsam RT, Kunapuli SP (2004) Central role of the P2Y12 receptor in platelet activation. J Clin Invest 113:340–345

    PubMed  CAS  Google Scholar 

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Acknowledgements

We thank Nethe Kristensen for technical support and Professor Ylva Hellsten and Professor Bengt Saltin (The Copenhagen Muscle Research Centre) for guidance and assistance. Novo Nordisk A/S financially supported the study.

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Authors and Affiliations

  1. Copenhagen Muscle Research Centre, Rigshospitalet, Section 7652, Tagensvej 29, DK-2200, Copenhagen N, Denmark

    A. Bornø, L. T. Bune & P. Thaning

  2. Department of Biomedical Sciences, Panum Institute, University of Copenhagen, Copenhagen, Denmark

    T. Ploug

  3. Department of Cardiology, Copenhagen University Hospital Gentofte, Copenhagen, Denmark

    J. B. Rosenmeier

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  1. A. Bornø
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Correspondence to P. Thaning.

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Bornø, A., Ploug, T., Bune, L.T. et al. Purinergic receptors expressed in human skeletal muscle fibres. Purinergic Signalling 8, 255–264 (2012). https://doi.org/10.1007/s11302-011-9279-y

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