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Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice

Abstract

The P2X7 receptor is an ATP-gated cation channel expressed by a number of cell types. We have shown previously that disruption of P2X7 receptor function results in downregulation of osteogenic markers and upregulation of adipogenic markers in calvarial cell cultures. In the present study, we assessed whether loss of P2X7 receptor function results in changes to adipocyte distribution and lipid accumulation in vivo. Male P2X7 loss-of-function (KO) mice exhibited significantly greater body weight and epididymal fat pad mass than wild-type (WT) mice at 9 months of age. Fat pad adipocytes did not differ in size, consistent with adipocyte hyperplasia rather than hypertrophy. Histological examination revealed ectopic lipid accumulation in the form of adipocytes and/or lipid droplets in several non-adipose tissues of older male KO mice (9–12 months of age). Ectopic lipid was observed in kidney, extraorbital lacrimal gland and pancreas, but not in liver, heart or skeletal muscle. Specifically, lacrimal gland and pancreas from 12-month-old male KO mice had greater numbers of adipocytes in perivascular, periductal and acinar regions. As well, lipid droplets accumulated in the renal tubular epithelium and lacrimal acinar cells. Blood plasma analyses revealed diminished total cholesterol levels in 9- and 12-month-old male KO mice compared with WT controls. Interestingly, no differences were observed in female mice. Moreover, there were no significant differences in food consumption between male KO and WT mice. Taken together, these data establish novel in vivo roles for the P2X7 receptor in regulating adipogenesis and lipid metabolism in an age- and sex-dependent manner.

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References

  1. 1.

    Dixon JB (2010) The effect of obesity on health outcomes. Mol Cell Endocrinol 316:104–108

  2. 2.

    Hassan M, Latif N, Yacoub M (2012) Adipose tissue: friend or foe? Nat Rev Cardiol 9:689–702

  3. 3.

    Dulloo AG, Montani JP (2012) Body composition, inflammation and thermogenesis in pathways to obesity and the metabolic syndrome: an overview. Obes Rev 13(Suppl 2):1–5

  4. 4.

    Shuldiner AR, Yang R, Gong DW (2001) Resistin, obesity and insulin resistance—the emerging role of the adipocyte as an endocrine organ. N Engl J Med 345:1345–1346

  5. 5.

    Ducy P, Amling M, Takeda S, Priemel M, Schilling AF, Beil FT, Shen J, Vinson C, Rueger JM, Karsenty G (2000) Leptin inhibits bone formation through a hypothalamic relay: a central control of bone mass. Cell 100:197–207

  6. 6.

    Ferron M, Wei J, Yoshizawa T, Del Fattore A, DePinho RA, Teti A, Ducy P, Karsenty G (2010) Insulin signaling in osteoblasts integrates bone remodeling and energy metabolism. Cell 142:296–308

  7. 7.

    Jacobi D, Stanya KJ, Lee CH (2012) Adipose tissue signaling by nuclear receptors in metabolic complications of obesity. Adipocyte 1:4–12

  8. 8.

    Suganami T, Tanaka M, Ogawa Y (2012) Adipose tissue inflammation and ectopic lipid accumulation. Endocr J 59:849–857

  9. 9.

    Kopecky J, Rossmeisl M, Flachs P, Brauner P, Sponarova J, Matejkova O, Prazak T, Ruzickova J, Bardova K, Kuda O (2004) Energy metabolism of adipose tissue—physiological aspects and target in obesity treatment. Physiol Res 53(Suppl 1):S225–S232

  10. 10.

    Lee NK, Sowa H, Hinoi E, Ferron M, Ahn JD, Confavreux C, Dacquin R, Mee PJ, McKee MD, Jung DY, Zhang Z, Kim JK, Mauvais-Jarvis F, Ducy P, Karsenty G (2007) Endocrine regulation of energy metabolism by the skeleton. Cell 130:456–469

  11. 11.

    Medina-Gomez G (2012) Mitochondria and endocrine function of adipose tissue. Best Pract Res Clin Endocrinol Metab 26:791–804

  12. 12.

    Burnstock G (2007) Physiology and pathophysiology of purinergic neurotransmission. Physiol Rev 87:659–797

  13. 13.

    Panupinthu N, Rogers JT, Zhao L, Solano-Flores LP, Possmayer F, Sims SM, Dixon SJ (2008) P2X7 receptors on osteoblasts couple to production of lysophosphatidic acid: a signaling axis promoting osteogenesis. J Cell Biol 181:859–871

  14. 14.

    Hughes JP, Hatcher JP, Chessell IP (2007) The role of P2X7 in pain and inflammation. Purinergic Signal 3:163–169

  15. 15.

    Sorge RE, Trang T, Dorfman R, Smith SB, Beggs S, Ritchie J, Austin JS, Zaykin DV, Vander Meulen H, Costigan M, Herbert TA, Yarkoni-Abitbul M, Tichauer D, Livneh J, Gershon E, Zheng M, Tan K, John SL, Slade GD, Jordan J, Woolf CJ, Peltz G, Maixner W, Diatchenko L, Seltzer Z, Salter MW, Mogil JS (2012) Genetically determined P2X7 receptor pore formation regulates variability in chronic pain sensitivity. Nat Med 18:595–599

  16. 16.

    Coutinho-Silva R, Persechini PM, Bisaggio RD, Perfettini JL, Neto AC, Kanellopoulos JM, Motta-Ly I, Dautry-Varsat A, Ojcius DM (1999) P2Z/P2X7 receptor-dependent apoptosis of dendritic cells. Am J Physiol 276:C1139–C1147

  17. 17.

    Gu BJ, Saunders BM, Petrou S, Wiley JS (2011) P2X7 is a scavenger receptor for apoptotic cells in the absence of its ligand, extracellular ATP. J Immunol 187:2365–2375

  18. 18.

    Surprenant A, Rassendren F, Kawashima E, North RA, Buell G (1996) The cytolytic P2Z receptor for extracellular ATP identified as a P2X receptor (P2X7). Science 272:735–738

  19. 19.

    Ke HZ, Qi H, Weidema AF, Zhang Q, Panupinthu N, Crawford DT, Grasser WA, Paralkar VM, Li M, Audoly LP, Gabel CA, Jee WS, Dixon SJ, Sims SM, Thompson DD (2003) Deletion of the P2X7 nucleotide receptor reveals its regulatory roles in bone formation and resorption. Mol Endocrinol 17:1356–1367

  20. 20.

    Li JLD, Ke HZ, Duncan RL, Turner CH (2005) The P2X7 nucleotide receptor mediates skeletal mechanotransduction. J Biol Chem 280:42952–42959

  21. 21.

    Grol MW, Panupinthu N, Korcok J, Sims SM, Dixon SJ (2009) Expression, signaling, and function of P2X7 receptors in bone. Purinergic Signal 5:205–221

  22. 22.

    Dubyak GR (2012) P2X7 receptor regulation of non-classical secretion from immune effector cells. Cell Microbiol 14:1697–1706

  23. 23.

    Miller CM, Boulter NR, Fuller SJ, Zakrzewski AM, Lees MP, Saunders BM, Wiley JS, Smith NC (2011) The role of the P2X7 receptor in infectious diseases. PLoS Pathog 7:e1002212

  24. 24.

    Sakowicz-Burkiewicz M, Kocbuch K, Grden M, Maciejewska I, Szutowicz A, Pawelczyk T (2013) High glucose concentration impairs ATP outflow and immunoglobulin production by human peripheral B lymphocytes: involvement of P2X7 receptor. Immunobiology 218:591–601

  25. 25.

    Solle M, Labasi J, Perregaux DG, Stam E, Petrushova N, Koller BH, Griffiths RJ, Gabel CA (2001) Altered cytokine production in mice lacking P2X7 receptors. J Biol Chem 276:125–132

  26. 26.

    Burnstock G, Novak I (2013) Purinergic signalling and diabetes. Purinergic Signal 9:307–324

  27. 27.

    Novak I (2008) Purinergic receptors in the endocrine and exocrine pancreas. Purinergic Signal 4:237–253

  28. 28.

    Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsenty G (1997) Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell 89:747–754

  29. 29.

    Rosen ED, Walkey CJ, Puigserver P, Spiegelman BM (2000) Transcriptional regulation of adipogenesis. Genes Dev 14:1293–1307

  30. 30.

    Muruganandan S, Roman AA, Sinal CJ (2009) Adipocyte differentiation of bone marrow-derived mesenchymal stem cells: cross talk with the osteoblastogenic program. Cell Mol Life Sci 66:236–253

  31. 31.

    Grol MW, Zelner I, Dixon SJ (2012) P2X7-mediated calcium influx triggers a sustained, PI3K-dependent increase in metabolic acid production by osteoblast-like cells. Am J Physiol Endocrinol Metab 302:E561–E575

  32. 32.

    Amoroso F, Falzoni S, Adinolfi E, Ferrari D, Di Virgilio F (2012) The P2X7 receptor is a key modulator of aerobic glycolysis. Cell Death Dis 3:e370

  33. 33.

    Sun S, Xia S, Ji Y, Kersten S, Qi L (2012) The ATP-P2X7 signaling axis is dispensable for obesity-associated inflammasome activation in adipose tissue. Diabetes 61:1471–1478

  34. 34.

    Madec S, Rossi C, Chiarugi M, Santini E, Salvati A, Ferrannini E, Solini A (2011) Adipocyte P2X7 receptors expression: a role in modulating inflammatory response in subjects with metabolic syndrome? Atherosclerosis 219:552–558

  35. 35.

    Sim JA, Young MT, Sung HY, North RA, Surprenant A (2004) Reanalysis of P2X7 receptor expression in rodent brain. J Neurosci 24:6307–6314

  36. 36.

    Labasi JM, Petrushova N, Donovan C, McCurdy S, Lira P, Payette MM, Brissette W, Wicks JR, Audoly L, Gabel CA (2002) Absence of the P2X7 receptor alters leukocyte function and attenuates an inflammatory response. J Immunol 168:6436–6445

  37. 37.

    Panupinthu N, Zhao L, Possmayer F, Ke HZ, Sims SM, Dixon SJ (2007) P2X7 nucleotide receptors mediate blebbing in osteoblasts through a pathway involving lysophosphatidic acid. J Biol Chem 282:3403–3412

  38. 38.

    Naemsch LN, Dixon SJ, Sims SM (2001) Activity-dependent development of P2X7 current and Ca2+ entry in rabbit osteoclasts. J Biol Chem 276:39107–39114

  39. 39.

    Masin M, Young C, Lim K, Barnes SJ, Xu XJ, Marschall V, Brutkowski W, Mooney ER, Gorecki DC, Murrell-Lagnado R (2012) Expression, assembly and function of novel C-terminal truncated variants of the mouse P2X7 receptor: re-evaluation of P2X7 knockouts. Br J Pharmacol 165:978–993

  40. 40.

    Granton PV, Norley CJ, Umoh J, Turley EA, Frier BC, Noble EG, Holdsworth DW (2010) Rapid in vivo whole body composition of rats using cone beam μCT. J Appl Physiol 109:1162–1169

  41. 41.

    Gulam M, Thornton MM, Hodsman AB, Holdsworth DW (2000) Bone mineral measurement of phalanges: comparison of radiographic absorptiometry and area dual X-ray absorptiometry. Radiology 216:586–591

  42. 42.

    Carson FL (1996) Histotechnology: a self-instructional text. American Society for Clinical Pathology, Chicago

  43. 43.

    Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226:497–509

  44. 44.

    Burnett JR, Wilcox LJ, Telford DE, Kleinstiver SJ, Barrett PH, Newton RS, Huff MW (1997) Inhibition of HMG-CoA reductase by atorvastatin decreases both VLDL and LDL apolipoprotein B production in miniature pigs. Arterioscler Thromb Vasc Biol 17:2589–2600

  45. 45.

    Hillman KA, Burnstock G, Unwin RJ (2005) The P2X7 ATP receptor in the kidney: a matter of life or death? Nephron Exp Nephrol 101:e24–e30

  46. 46.

    Belazi D, Sole-Domenech S, Johansson B, Schalling M, Sjovall P (2009) Chemical analysis of osmium tetroxide staining in adipose tissue using imaging ToF-SIMS. Histochem Cell Biol 132:105–115

  47. 47.

    Hodges RR, Vrouvlianis J, Shatos MA, Dartt DA (2009) Characterization of P2X7 purinergic receptors and their function in rat lacrimal gland. Invest Ophthalmol Vis Sci 50:5681–5689

  48. 48.

    Pochet S, Garcia-Marcos M, Seil M, Otto A, Marino A, Dehaye JP (2007) Contribution of two ionotropic purinergic receptors to ATP responses in submandibular gland ductal cells. Cell Signal 19:2155–2164

  49. 49.

    Nakamoto T, Brown DA, Catalan MA, Gonzalez-Begne M, Romanenko VG, Melvin JE (2009) Purinergic P2X7 receptors mediate ATP-induced saliva secretion by the mouse submandibular gland. J Biol Chem 284:4815–4822

  50. 50.

    Costa-Junior HM, Marques-da-Silva C, Vieira FS, Moncao-Ribeiro LC, Coutinho-Silva R (2011) Lipid metabolism modulation by the P2X7 receptor in the immune system and during the course of infection: new insights into the old view. Purinergic Signal 7:381–392

  51. 51.

    Taylor SR, Turner CM, Elliott JI, McDaid J, Hewitt R, Smith J, Pickering MC, Whitehouse DL, Cook HT, Burnstock G, Pusey CD, Unwin RJ, Tam FW (2009) P2X7 deficiency attenuates renal injury in experimental glomerulonephritis. J Am Soc Nephrol 20:1275–1281

  52. 52.

    Bjorntorp P (1991) Adipose tissue distribution and function. Int J Obes 15(Suppl 2):67–81

  53. 53.

    Volonte C, Amadio S, D’Ambrosi N, Colpi M, Burnstock G (2006) P2 receptor web: complexity and fine-tuning. Pharmacol Ther 112:264–280

  54. 54.

    Drolet R, Richard C, Sniderman AD, Mailloux J, Fortier M, Huot C, Rheaume C, Tchernof A (2008) Hypertrophy and hyperplasia of abdominal adipose tissues in women. Int J Obes (Lond) 32:283–291

  55. 55.

    Bouacida A, Rosset P, Trichet V, Guilloton F, Espagnolle N, Cordonier T, Heymann D, Layrolle P, Sensebe L, Deschaseaux F (2012) Pericyte-like progenitors show high immaturity and engraftment potential as compared with mesenchymal stem cells. PLoS One 7:e48648

  56. 56.

    Zippel N, Limbach CA, Ratajski N, Urban C, Luparello C, Pansky A, Kassack MU, Tobiasch E (2012) Purinergic receptors influence the differentiation of human mesenchymal stem cells. Stem Cells Dev 21:884–900

  57. 57.

    You S, Kublin CL, Avidan O, Miyasaki D, Zoukhri D (2011) Isolation and propagation of mesenchymal stem cells from the lacrimal gland. Invest Ophthalmol Vis Sci 52:2087–2094

  58. 58.

    Rovira M, Scott SG, Liss AS, Jensen J, Thayer SP, Leach SD (2010) Isolation and characterization of centroacinar/terminal ductal progenitor cells in adult mouse pancreas. Proc Natl Acad Sci U S A 107:75–80

  59. 59.

    Le Stunff H, Auger R, Kanellopoulos J, Raymond MN (2004) The Pro-451 to Leu polymorphism within the C-terminal tail of P2X7 receptor impairs cell death but not phospholipase D activation in murine thymocytes. J Biol Chem 279:16918–16926

  60. 60.

    Novak I, Jans IM, Wohlfahrt L (2010) Effect of P2X7 receptor knockout on exocrine secretion of pancreas, salivary glands and lacrimal glands. J Physiol 588:3615–3627

  61. 61.

    Cario-Toumaniantz C, Loirand G, Ferrier L, Pacaud P (1998) Non-genomic inhibition of human P2X7 purinoceptor by 17β-oestradiol. J Physiol 508:659–666

  62. 62.

    Hillman KA, Johnson TM, Winyard PJ, Burnstock G, Unwin RJ, Woolf AS (2002) P2X7 receptors are expressed during mouse nephrogenesis and in collecting duct cysts of the cpk/cpk mouse. Exp Nephrol 10:34–42

  63. 63.

    Emmett DS, Feranchak A, Kilic G, Puljak L, Miller B, Dolovcak S, McWilliams R, Doctor RB, Fitz JG (2008) Characterization of ionotrophic purinergic receptors in hepatocytes. Hepatology 47:698–705

  64. 64.

    Young CN, Brutkowski W, Lien CF, Arkle S, Lochmuller H, Zablocki K, Gorecki DC (2012) P2X7 purinoceptor alterations in dystrophic mdx mouse muscles: relationship to pathology and potential target for treatment. J Cell Mol Med 16:1026–1037

  65. 65.

    Daugherty A (2002) Mouse models of atherosclerosis. Am J Med Sci 323:3–10

  66. 66.

    Zadelaar S, Kleemann R, Verschuren L, de Vries-Van der Weij J, van der Hoorn J, Princen HM, Kooistra T (2007) Mouse models for atherosclerosis and pharmaceutical modifiers. Arterioscler Thromb Vasc Biol 27:1706–1721

  67. 67.

    Getz GS, Reardon CA (2012) Animal models of atherosclerosis. Arterioscler Thromb Vasc Biol 32:1104–1115

  68. 68.

    Basso AM, Bratcher NA, Harris RR, Jarvis MF, Decker MW, Rueter LE (2009) Behavioral profile of P2X7 receptor knockout mice in animal models of depression and anxiety: relevance for neuropsychiatric disorders. Behav Brain Res 198:83–90

  69. 69.

    Glas R, Sauter NS, Schulthess FT, Shu L, Oberholzer J, Maedler K (2009) Purinergic P2X7 receptors regulate secretion of interleukin-1 receptor antagonist and beta cell function and survival. Diabetologia 52:1579–1588

  70. 70.

    Houtkooper RH, Argmann C, Houten SM, Canto C, Jeninga EH, Andreux PA, Thomas C, Doenlen R, Schoonjans K, Auwerx J (2011) The metabolic footprint of aging in mice. Sci Rep 1:134

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Acknowledgements

This work was funded by the Canadian Institutes of Health Research (CIHR) [grant number 102542]. KLB and AX were supported in part by the Joint Motion Program—A CIHR Training Program in Musculoskeletal Health Research and Leadership. MWG was supported by a CIHR Frederick Banting and Charles Best Canada Graduate Scholarship Doctoral Award. DWH holds the Dr. Sandy Kirkley Chair in Musculoskeletal Research at The University of Western Ontario. We thank Dr. Nattapon Panupinthu for performing preliminary studies, Linda Jackson and Tom Chrones for assistance with histology, Vasek Pitelka for aiding with animal anaesthesia, Dr. Joseph Umoh for micro-CT scanning, Drs. Tom Daley and Ian Welch for advice on histopathology, Cindy Sawyez and Brian Sutherland for assistance with biochemical analyses and Dr. Nica Borradaile for helpful comments.

Conflict of interest

The authors have no conflicts of interest to declare.

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Correspondence to S. Jeffrey Dixon.

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Beaucage, K.L., Xiao, A., Pollmann, S.I. et al. Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice. Purinergic Signalling 10, 291–304 (2014). https://doi.org/10.1007/s11302-013-9388-x

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Keywords

  • Adipocyte
  • Cholesterol
  • Exocrine
  • Kidney
  • Metabolic syndrome
  • P2rx7