In vivo radiometric analysis of glucose uptake and distribution in mouse bone

Abstract

Bone formation and remodeling occurs throughout life and requires the sustained activity of osteoblasts and osteoclasts, particularly during periods of rapid bone growth. Despite increasing evidence linking bone cell activity to global energy homeostasis, little is known about the relative energy requirements or substrate utilization of bone cells. In these studies, we measured the uptake and distribution of glucose in the skeleton in vivo using positron-emitting ¹⁸F-fluorodeoxyglucose ([¹⁸F]-FDG) and non-invasive, high-resolution positron emission tomography/computed tomography (PET/CT) imaging and ex vivo autoradiography. Assessment of [¹⁸F]-FDG uptake demonstrated that relative to other tissues bone accumulated a significant fraction of the total dose of the glucose analog. Skeletal accumulation was greatest in young mice undergoing the rapid bone formation that characterizes early development. PET/CT imaging revealed that [¹⁸F]-FDG uptake was greatest in the epiphyseal and metaphyseal regions of long bones, which accords with the increased osteoblast numbers and activity at this skeletal site. Insulin administration significantly increased skeletal accumulation of [¹⁸F]-FDG, while uptake was reduced in mice lacking the insulin receptor specifically in osteoblasts or fed a high-fat diet. Our results indicated that the skeleton is a site of significant glucose uptake and that its consumption by bone cells is subject to regulation by insulin and disturbances in whole-body metabolism.

Metabolism: Glucose uptake and distribution in bones

The skeleton is a site of significant glucose uptake, which is regulated by insulin and affected by whole body metabolic changes. According to Ryan Riddle and DanielThorek from the Johns Hopkins University School of Medicine, Baltimore, US, and colleagues, little is known about the energy requirements of bone cells,despite evidence linking bone growth to energy intake. Using radiolabelled glucose analogs in mice, they found evidence of glucose uptake within the bone cells, particularly within spongy bone. The extent of glucose uptake exceeded that of other glucose-storing organs such as the liver and muscle. The uptake appeared to be regulated by insulin. The findings suggest that the skeleton should be considered as a possible site of significant glucose disposal and a major component of whole body glucose metabolism.

References

[1] Buttgereit F, Brand MD. A hierarchy of ATP-consuming processes in mammalian cells. Biochem J, 1995, 312 Pt 1 163-167

[2] Passi-Even L, Gazit D, Bab I. Ontogenesis of ultrastructural features during osteogenic differentiation in diffusion chamber cultures of marrow cells. J Bone Miner Res, 1993, 8: 589-595

[3] Komarova SV, Ataullakhanov FI, Globus RK. Bioenergetics and mitochondrial transmembrane potential during differentiation of cultured osteoblasts. Am J Physiol Cell Physiol, 2000, 279: C1220-C1229

[4] Klein BY, Gal I, Hartshtark Z et al Induction of osteoprogenitor cell differentiation in rat marrow stroma increases mitochondrial retention of rhodamine 123 in stromal cells. J Cell Biochem, 1993, 53: 190-197

[5] Esen E, Chen J, Karner CM et al WNT-LRP5 signaling induces Warburg effect through mTORC2 activation during osteoblast differentiation. Cell Metabolism, 2013, 17: 745-755

[6] Guntur AR, Le PT, Farber CR et al Bioenergetics during calvarial osteoblast differentiation reflect strain differences in bone mass. Endocrinology, 2014, 155: 1589-1595

[7] DiGirolamo DJ, Clemens TL, Kousteni S. The skeleton as an endocrine organ. Nat Rev Rheumatol, 2012, 8: 674-683

[8] Lee NK, Sowa H, Hinoi E et al Endocrine regulation of energy metabolism by the skeleton. Cell, 2007, 130: 456-469

[9] Zoch ML, Clemens TL, Riddle RC. New insights into the biology of osteocalcin. Bone, 2016, 82: 42-49

[10] Karsenty G, Ferron M. The contribution of bone to whole-organism physiology. Nature, 2012, 481: 314-320

[11] Ferron M, Hinoi E, Karsenty G et al Osteocalcin differentially regulates beta cell and adipocyte gene expression and affects the development of metabolic diseases in wild-type mice. Proc Natl Acad Sci USA, 2008, 105: 5266-5270

[12] Ferron M, McKee MD, Levine RL et al Intermittent injections of osteocalcin improve glucose metabolism and prevent type 2 diabetes in mice. Bone, 2011, 50: 568-575

[13] Ferron M, Wei J, Yoshizawa T et al Insulin signaling in osteoblasts integrates bone remodeling and energy metabolism. Cell, 2010, 142: 296-308

[14] Fulzele K, Riddle RC, DiGirolamo DJ et al Insulin receptor signaling in osteoblasts regulates postnatal bone acquisition and body composition. Cell, 2010, 142: 309-319

[15] Yoshikawa Y, Kode A, Xu L et al Genetic evidence points to an osteocalcin-independent influence of osteoblasts on energy metabolism. J Bone Miner Res, 2011, 26: 2012-2025

[16] Borle AB, Nichols N, Nichols G Jr. Metabolic studies of bone in vitro. I. Normal bone. J Biol Chem, 1960, 235: 1206-1210

[17] Cohn DV, Forscher BK. Aerobic metabolism of glucose by bone. J Biol Chem, 1962, 237: 615-618

[18] Peck WA, Birge SJ Jr, Fedak SA. Bone Cells: Biochemical and Biological Studies after Enzymatic Isolation. Science, 1964, 146: 1476-1477

[19] Wei J, Shimazu J, Makinistoglu MP et al Glucose uptake and Runx2 synergize to orchestrate osteoblast differentiation and bone formation. Cell, 2015, 161: 1576-1591

[20] Thomas DM, Rogers SD, Sleeman MW et al Modulation of glucose transport by parathyroid hormone and insulin in UMR 106-01, a clonal rat osteogenic sarcoma cell line. J Mol Endocrinol, 1995, 14: 263-275

[21] Esen E, Lee SY, Wice BM et al PTH Promotes bone anabolism by stimulating aerobic glycolysis via IGF signaling. J Bone Miner Res, 2015, 30: 2137

[22] Ituarte EA, Halstead LR, Iida-Klein A et al Glucose transport system in UMR-106-01 osteoblastic osteosarcoma cells: regulation by insulin. Calcif Tissue Int, 1989, 45: 27-33

[23] Zoidis E, Ghirlanda-Keller C, Schmid C. Stimulation of glucose transport in osteoblastic cells by parathyroid hormone and insulin-like growth factor I. Mol Cell Biochem, 2011, 348: 33-42

[24] Karner CM, Esen E, Okunade AL et al Increased glutamine catabolism mediates bone anabolism in response to WNT signaling. J Clin Invest, 2015, 125: 551-562

[25] Frey JL, Li Z, Ellis JM et al Wnt-Lrp5 signaling regulates fatty acid metabolism in the osteoblast. Mol Cell Biol, 2015, 35: 1979-1991

[26] Adamek G, Felix R, Guenther HL et al Fatty acid oxidation in bone tissue and bone cells in culture. Characterization and hormonal influences. Biochem J, 1987, 248: 129-137

[27] Niemeier A, Niedzielska D, Secer R et al Uptake of postprandial lipoproteins into bone in vivo: impact on osteoblast function. Bone, 2008, 43: 230-237

[28] Salas J, Salas M, Vinuela E et al Glucokinase of rabbit liver. J Biol Chem, 1965, 240: 1014-1018

[29] Sokoloff L, Reivich M, Kennedy C et al The [¹⁴C]deoxyglucose method for the measurement of local cerebral glucose utilization: theory, procedure, and normal values in the conscious and anesthetized albino rat. J Neurochem, 1977, 28: 897-916

[30] Wick AN, Drury DR, Nakada HI et al Localization of the primary metabolic block produced by 2-deoxyglucose. J Biol Chem, 1957, 224: 963-969

[31] Phelps ME, Huang SC, Hoffman EJ et al Tomographic measurement of local cerebral glucose metabolic rate in humans with (F-18)2-fluoro-2-deoxy-D-glucose: validation of method. Ann Neurol, 1979, 6: 371-388

[32] Bruning JC, Michael MD, Winnay JN et al A muscle-specific insulin receptor knockout exhibits features of the metabolic syndrome of NIDDM without altering glucose tolerance. Mol Cell, 1998, 2: 559-569

[33] Zhang M, Xuan S, Bouxsein ML et al Osteoblast-specific knockout of the insulin-like growth factor (IGF) receptor gene reveals an essential role of IGF signaling in bone matrix mineralization. J Biol Chem, 2002, 277: 44005-44012

[34] Bender D, Munk OL, Feng HQ et al Metabolites of (18)F-FDG and 3-O-(11)C-methylglucose in pig liver. J Nucl Med, 2001, 42: 1673-1678

[35] Gallagher BM, Fowler JS, Gutterson NI et al Metabolic trapping as a principle of oradiopharmaceutical design: some factors resposible for the biodistribution of [¹⁸F] 2-deoxy-2-fluoro-D-glucose. J Nucl Med, 1978, 19: 1154-1161

[36] Fueger BJ, Czernin J, Hildebrandt I et al Impact of animal handling on the results of ¹⁸F-FDG PET studies in mice. J Nucl Med, 2006, 47: 999-1006

[37] Wei J, Ferron M, Clarke CJ et al Bone-specific insulin resistance disrupts whole-body glucose homeostasis via decreased osteocalcin activation. J Clin Invest, 2014, 124: 1-13

[38] Thiebaud D, Jacot E, DeFronzo RA et al The effect of graded doses of insulin on total glucose uptake, glucose oxidation, and glucose storage in man. Diabetes, 1982, 31: 957-963

[39] Kim JK, Michael MD, Previs SF et al Redistribution of substrates to adipose tissue promotes obesity in mice with selective insulin resistance in muscle. J Clin Invest, 2000, 105: 1791-1797

[40] Richter EA, Hargreaves M. Exercise, GLUT4, and skeletal muscle glucose uptake. Physiol Rev, 2013, 93: 993-1017

[41] Skerry TM, Bitensky L, Chayen J et al Early strain-related changes in enzyme activity in osteocytes following bone loading in vivo. J Bone Miner Res, 1989, 4: 783-788

[42] Beamer WG, Donahue LR, Rosen CJ et al Genetic variability in adult bone density among inbred strains of mice. Bone, 1996, 18: 397-403

[43] Glatt V, Canalis E, Stadmeyer L et al Age-related changes in trabecular architecture differ in female and male C57BL/6J mice. J Bone Miner Res, 2007, 22: 1197-1207