In vivo high-resolution magic angle spinning proton NMR spectroscopy of Drosophila melanogaster flies as a model system to investigate mitochondrial dysfunction in Drosophila GST2 mutants

Righi,Valeria; Apidianakis,Yiorgos; Psychogios,Nikolaos; Rahme,Laurence G.; Tompkins,Ronald G.; Tzika,A. Aria

doi:10.3892/ijmm.2014.1757

In vivo high-resolution magic angle spinning proton NMR spectroscopy of Drosophila melanogaster flies as a model system to investigate mitochondrial dysfunction in Drosophila GST2 mutants

Authors:
- Valeria Righi
- Yiorgos Apidianakis
- Nikolaos Psychogios
- Laurence G. Rahme
- Ronald G. Tompkins
- A. Aria Tzika
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Affiliations: NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital and Shriners Burn Institute, Harvard Medical School, Boston, MA 02114, USA , Division of Burns, Department of Surgery, Massachusetts General Hospital and Shriners Burns Institute, Harvard Medical School, Boston, MA 02114, USA
Published online on: April 24, 2014 https://doi.org/10.3892/ijmm.2014.1757
Pages: 327-333

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Abstract

In vivo nuclear magnetic resonance spectroscopy (NMR), a non-destructive biochemical tool used for investigating live organisms, has recently been performed in studies of the fruit fly Drosophila melanogaster, a useful model organism for investigating genetics and physiology. We used a novel high-resolution magic angle-spinning (HRMAS) NMR method to investigate live Drosophila GST2 mutants using a conventional 14.1-T NMR spectrometer equipped with an HRMAS probe. The results showed that, compared to wild-type (wt) controls, the GST2 mutants had a 48% greater (CH2)n lipid signal at 1.33 ppm, which is an insulin resistance biomarker in Drosophila skeletal muscle (P=0.0444). The mutants also had a 57% greater CH2C= lipid signal at 2.02 ppm (P=0.0276) and a 100% greater -CH=CH- signal at 5.33 ppm (P=0.0251). Since the -CH=CH- signal encompasses protons from ceramide, this latter difference is consistent with the hypothesis that the GST2 mutation is associated with insulin resistance and apoptosis. The findings of this study corroborate our previous results, support the hypothesis that the GST2 mutation is associated with insulin signaling and suggest that the IMCL level may be a biomarker of insulin resistance. Furthermore, direct links between GST2 mutation (the Drosophila ortholog of the GSTA4 gene in mammals) and insulin resistance, as suggested in this study, have not been made previously. These findings may thus be directly relevant to a wide range of metabolically disruptive conditions, such as trauma, aging and immune system deficiencies, that lead to increased susceptibility to infection.

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1	Weybright P, Millis K, Campbell N, Cory DG and Singer S: Gradient, high-resolution, magic angle spinning ¹H nuclear magnetic resonance spectroscopy of intact cells. Magn Reson Med. 39:337–345. 1998. View Article : Google Scholar : PubMed/NCBI
2	Blankenberg FG, Storrs RW, Naumovski L, Goralski T and Spielman D: Detection of apoptotic cell death by proton nuclear magnetic resonance spectroscopy. Blood. 87:1951–1956. 1996.PubMed/NCBI
3	Cheng LL, Ma MJ, Becerra L, et al: Quantitative neuropathology by high resolution magic angle spinning proton magnetic resonance spectroscopy. Proc Natl Acad Sci USA. 94:6408–6413. 1997. View Article : Google Scholar : PubMed/NCBI
4	Cheng LL, Newell K, Mallory AE, Hyman BT and Gonzalez RG: Quantification of neurons in Alzheimer and control brains with ex vivo high resolution magic angle spinning proton magnetic resonance spectroscopy and stereology. Magn Reson Imaging. 20:527–533. 2002. View Article : Google Scholar
5	Millis KK, Maas WE, Cory DG and Singer S: Gradient, high-resolution, magic-angle spinning nuclear magnetic resonance spectroscopy of human adipocyte tissue. Magn Reson Med. 38:399–403. 1997. View Article : Google Scholar : PubMed/NCBI
6	Millis K, Weybright P, Campbell N, et al: Classification of human liposarcoma and lipoma using ex vivo proton NMR spectroscopy. Magn Reson Med. 41:257–267. 1999. View Article : Google Scholar : PubMed/NCBI
7	Barton SJ, Howe FA, Tomlins AM, et al: Comparison of in vivo ¹H MRS of human brain tumours with ¹H HR-MAS spectroscopy of intact biopsy samples in vitro. MAGMA. 8:121–128. 1999.
8	Griffin JL, Williams HJ, Sang E and Nicholson JK: Abnormal lipid profile of dystrophic cardiac tissue as demonstrated by one- and two-dimensional magic-angle spinning (1)H NMR spectroscopy. Magn Reson Med. 46:249–255. 2001. View Article : Google Scholar
9	Tzika AA, Cheng LL, Goumnerova L, et al: Biochemical characterization of pediatric brain tumors by using in vivo and ex vivo magnetic resonance spectroscopy. J Neurosurg. 96:1023–1031. 2002. View Article : Google Scholar : PubMed/NCBI
10	Tugnoli V, Schenetti L, Mucci A, et al: Ex vivo HR-MAS MRS of human meningiomas: a comparison with in vivo ¹H MR spectra. Int J Mol Med. 18:859–869. 2006.
11	Astrakas LG, Goljer I, Yasuhara S, et al: Proton NMR spectroscopy shows lipids accumulate in skeletal muscle in response to burn trauma-induced apoptosis. FASEB J. 19:1431–1440. 2005. View Article : Google Scholar : PubMed/NCBI
12	Tzika AA, Astrakas LG, Cao H, et al: Murine intramyocellular lipids quantified by NMR act as metabolic biomarkers in burn trauma. Int J Mol Med. 21:825–832. 2008.PubMed/NCBI
13	Bollard ME, Garrod S, Holmes E, et al: High-resolution (1)H and (1)H-(13)C magic angle spinning NMR spectroscopy of rat liver. Magn Reson Med. 44:201–207. 2000. View Article : Google Scholar : PubMed/NCBI
14	Szczepaniak LS, Babcock EE, Schick F, et al: Measurement of intracellular triglyceride stores by H spectroscopy: validation in vivo. Am J Physiol. 276:E977–E989. 1999.PubMed/NCBI
15	van der Graaf M, Tack CJ, de Haan JH, Klomp DW and Heerschap A: Magnetic resonance spectroscopy shows an inverse correlation between intramyocellular lipid content in human calf muscle and local glycogen synthesis rate. NMR Biomed. 23:133–141. 2009.
16	Jacob S, Machann J, Rett K, et al: Association of increased intramyocellular lipid content with insulin resistance in lean nondiabetic offspring of type 2 diabetic subjects. Diabetes. 48:1113–1119. 1999. View Article : Google Scholar : PubMed/NCBI
17	Petersen KF, Befroy D, Dufour S, et al: Mitochondrial dysfunction in the elderly: possible role in insulin resistance. Science. 300:1140–1142. 2003. View Article : Google Scholar : PubMed/NCBI
18	Feala JD, Coquin L, McCulloch AD and Paternostro G: Flexibility in energy metabolism supports hypoxia tolerance in Drosophila flight muscle: metabolomic and computational systems analysis. Mol Syst Biol. 3:992007. View Article : Google Scholar : PubMed/NCBI
19	Pedersen KS, Kristensen TN, Loeschcke V, et al: Metabolomic signatures of inbreeding at benign and stressful temperatures in Drosophila melanogaster. Genetics. 180:1233–1243. 2008. View Article : Google Scholar : PubMed/NCBI
20	Bharucha KN: The epicurean fly: using Drosophila melanogaster to study metabolism. Pediatr Res. 65:132–137. 2009.PubMed/NCBI
21	Null B, Liu CW, Hedehus M, Conolly S and Davis RW: High-resolution, in vivo magnetic resonance imaging of Drosophila at 18.8 Tesla. PLoS One. 3:e28172008. View Article : Google Scholar : PubMed/NCBI
22	Righi V, Apidianakis Y, Rahme LG and Tzika AA: Magnetic resonance spectroscopy of live Drosophila melanogaster using magic angle spinning. J Vis Exp. 38:17102010.
23	Baker KD and Thummel CS: Diabetic larvae and obese flies-emerging studies of metabolism in Drosophila. Cell Metab. 6:257–266. 2007. View Article : Google Scholar : PubMed/NCBI
24	Leopold P and Perrimon N: Drosophila and the genetics of the internal milieu. Nature. 450:186–188. 2007. View Article : Google Scholar
25	Singh SP, Coronella JA, Benes H, Cochrane BJ and Zimniak P: Catalytic function of Drosophila melanogaster glutathione S-transferase DmGSTS1–1 (GST-2) in conjugation of lipid peroxidation end products. Eur J Biochem. 268:2912–2923. 2001.PubMed/NCBI
26	Meiboom S and Gill D: Modified spiin-echo method for measuring nuclear relaxation time. Rev Sci Instrum. 29:688–691. 1958. View Article : Google Scholar
27	Levenberg K: A method for the solution of certain non-linear problems in least squares. Q Appl Math. 2:164–168. 1944.
28	Marquardt D: An algorithm for least-squares estimation of nonlinear parameters. SIAM J Appl Math. 11:431–441. 1963. View Article : Google Scholar
29	Swanson MG, Zektzer AS, Tabatabai ZL, et al: Quantitative analysis of prostate metabolites using ¹H HR-MAS spectroscopy. Magn Reson Med. 55:1257–1264. 2006. View Article : Google Scholar : PubMed/NCBI
30	Righi V, Apidianakis Y, Psychogios N, Rahme LG, Tompkins RG and Tzika AA: In vivo high-resolution magic angle spinning proton NMR spectroscopy of Drosophila melanogaster flies as a model system to investigate mitochondrial dysfunction in Drosophila mutants. Intl Soc Mag Reson Med. 1460:192011.
31	Garofalo RS: Genetic analysis of insulin signaling in Drosophila. Trends Endocrinol Metab. 13:156–162. 2002. View Article : Google Scholar : PubMed/NCBI
32	Saltiel AR and Kahn CR: Insulin signalling and the regulation of glucose and lipid metabolism. Nature. 414:799–806. 2001. View Article : Google Scholar : PubMed/NCBI
33	Abmayr SM, Zhuang S and Geisbrecht ER: Myoblast fusion in Drosophila. Methods Mol Biol. 475:75–97. 2008. View Article : Google Scholar
34	Richardson B, Beckett K and Baylies M: Visualizing new dimensions in Drosophila myoblast fusion. Bioessays. 30:423–431. 2008. View Article : Google Scholar : PubMed/NCBI
35	Rochlin K, Yu S, Roy S and Baylies MK: Myoblast fusion: when it takes more to make one. Dev Biol. 341:66–83. 2010. View Article : Google Scholar : PubMed/NCBI
36	Partridge L and Tower J: Yeast, a feast: The fruit fly Drosophila as a model organism for research into aging. The Molecular Biology of Aging. Guarente L and Partridge L: Cold Spring Harbor Laboratory Press; pp. 267–308. 2008
37	Marsh JL and Thompson LM: Drosophila in the study of neurodegenerative disease. Neuron. 52:169–178. 2006. View Article : Google Scholar
38	Ramsden S, Cheung YY and Seroude L: Functional analysis of the Drosophila immune response during aging. Aging Cell. 7:225–236. 2008.
39	Zerofsky M, Harel E, Silverman N and Tatar M: Aging of the innate immune response in Drosophila melanogaster. Aging Cell. 4:103–108. 2005.PubMed/NCBI
40	Ocorr K, Akasaka T and Bodmer R: Age-related cardiac disease model of Drosophila. Mech Ageing Dev. 128:112–116. 2007. View Article : Google Scholar : PubMed/NCBI
41	Smith JM, Bozcuk AN and Tebbutt S: Protein turnover in adult Drosophila. J Insect Physiol. 16:601–613. 1970. View Article : Google Scholar
42	Webster GC, Beachell VT and Webster SL: Differential decrease in protein synthesis by microsomes from aging Drosophila melanogaster. Exp Gerontol. 15:495–497. 1980. View Article : Google Scholar : PubMed/NCBI
43	Gartner LP: Aging and the visceral musculature of the adult fruitfly: an ultrastructural investigation. Trans Am Microsc Soc. 96:48–55. 1977. View Article : Google Scholar : PubMed/NCBI
44	Miller MS, Lekkas P, Braddock JM, et al: Aging enhances indirect flight muscle fiber performance yet decreases flight ability in Drosophila. Biophys J. 95:2391–2401. 2008. View Article : Google Scholar : PubMed/NCBI
45	Takahashi A, Philpott DE and Miquel J: Electron microscope studies on aging Drosophila melanogaster. 3 Flight muscle. J Gerontol. 25:222–228. 1970. View Article : Google Scholar
46	Zheng J, Edelman SW, Tharmarajah G, Walker DW, Pletcher SD and Seroude L: Differential patterns of apoptosis in response to aging in Drosophila. Proc Natl Acad Sci USA. 102:12083–12088. 2005. View Article : Google Scholar : PubMed/NCBI
47	Ferguson M, Mockett RJ, Shen Y, Orr WC and Sohal RS: Age-associated decline in mitochondrial respiration and electron transport in Drosophila melanogaster. Biochem J. 390:501–511. 2005. View Article : Google Scholar : PubMed/NCBI
48	Girardot F, Lasbleiz C, Monnier V and Tricoire H: Specific age-related signatures in Drosophila body parts transcriptome. BMC Genomics. 7:692006. View Article : Google Scholar : PubMed/NCBI
49	Magwere T, Goodall S, Skepper J, Mair W, Brand MD and Partridge L: The effect of dietary restriction on mitochondrial protein density and flight muscle mitochondrial morphology in Drosophila. J Gerontol A Biol Sci Med Sci. 61:36–47. 2006. View Article : Google Scholar : PubMed/NCBI
50	Sohal RS, Sohal BH and Orr WC: Mitochondrial superoxide and hydrogen peroxide generation, protein oxidative damage, and longevity in different species of flies. Free Radic Biol Med. 19:499–504. 1995. View Article : Google Scholar : PubMed/NCBI
51	Goddeeris MM, Cook-Wiens E, Horton WJ, et al: Delayed behavioural aging and altered mortality in Drosophila beta integrin mutants. Aging Cell. 2:257–264. 2003.PubMed/NCBI
52	Miller BM, Zhang S, Suggs JA, et al: An alternative domain near the nucleotide-binding site of Drosophila muscle myosin affects ATPase kinetics. J Mol Biol. 353:14–25. 2005. View Article : Google Scholar : PubMed/NCBI
53	Kronert WA, Dambacher CM, Knowles AF, Swank DM and Bernstein SI: Alternative relay domains of Drosophila melanogaster myosin differentially affect ATPase activity, in vitro motility, myofibril structure and muscle function. J Mol Biol. 379:443–456. 2008.PubMed/NCBI
54	Kronert WA, Melkani GC, Melkani A and Bernstein SI: Mutating the converter-relay interface of Drosophila myosin perturbs ATPase activity, actin motility, myofibril stability and flight ability. J Mol Biol. 398:625–632. 2010.PubMed/NCBI
55	Das N, Levine RL, Orr WC and Sohal RS: Selectivity of protein oxidative damage during aging in Drosophila melanogaster. Biochem J. 360:209–216. 2001. View Article : Google Scholar : PubMed/NCBI
56	Toroser D, Orr WC and Sohal RS: Carbonylation of mitochondrial proteins in Drosophila melanogaster during aging. Biochem Biophys Res Commun. 363:418–424. 2007. View Article : Google Scholar : PubMed/NCBI
57	Wheeler JC, Bieschke ET and Tower J: Muscle-specific expression of Drosophila hsp70 in response to aging and oxidative stress. Proc Natl Acad Sci USA. 92:10408–10412. 1995.
58	Zhan M, Yamaza H, Sun Y, Sinclair J, Li H and Zou S: Temporal and spatial transcriptional profiles of aging in Drosophila melanogaster. Genome Res. 17:1236–1243. 2007. View Article : Google Scholar : PubMed/NCBI
59	Singh SP, Niemczyk M, Saini D, Awasthi YC, Zimniak L and Zimniak P: Role of the electrophilic lipid peroxidation product 4-hydroxynonenal in the development and maintenance of obesity in mice. Biochemistry. 47:3900–3911. 2008. View Article : Google Scholar : PubMed/NCBI
60	Weis J, Johansson L, Ortiz-Nieto F and Ahlstrom H: Assessment of lipids in skeletal muscle by LCModel and AMARES. J Magn Reson Imaging. 30:1124–1129. 2009. View Article : Google Scholar : PubMed/NCBI
61	Wang L, Salibi N, Wu Y, Schweitzer ME and Regatte RR: Relaxation times of skeletal muscle metabolites at 7T. J Magn Reson Imaging. 29:1457–1464. 2009. View Article : Google Scholar : PubMed/NCBI
62	Chen JH, Sambol EB, Decarolis P, et al: High-resolution MAS NMR spectroscopy detection of the spin magnetization exchange by cross-relaxation and chemical exchange in intact cell lines and human tissue specimens. Magn Reson Med. 55:1246–1256. 2006. View Article : Google Scholar : PubMed/NCBI
63	Boesch C, Slotboom J, Hoppeler H and Kreis R: In vivo determination of intra-myocellular lipids in human muscle by means of localized ¹H-MR-spectroscopy. Magn Reson Med. 37:484–493. 1997. View Article : Google Scholar : PubMed/NCBI
64	Vermathen P, Kreis R and Boesch C: Distribution of intramyocellular lipids in human calf muscles as determined by MR spectroscopic imaging. Magn Reson Med. 51:253–262. 2004. View Article : Google Scholar : PubMed/NCBI
65	Havel RJ, Carlson LA, Ekelund LG and Holmgren A: Turnover rate and oxidation of different free fatty acids in man during exercise. J Appl Physiol. 19:613–618. 1964.PubMed/NCBI
66	Mehring M: High Resolution NMR in Solids. Springer-Verlag; New York: 1982
67	Garroway AN: Magic-angle sample spinning of liquids. J Magn Reson. 49:168–171. 1982.
68	Barbara TM: Cylindrical demagnetization fields and microprobe design in high resolution NMR. J Magn Reson A. 109:2651994. View Article : Google Scholar
69	Chen JH, Enloe BM, Xiao Y, Cory DG and Singer S: Isotropic susceptibility shift under MAS: the origin of the split water resonances in ¹H MAS NMR spectra of cell suspensions. Magn Reson Med. 50:515–521. 2003. View Article : Google Scholar : PubMed/NCBI
70	Chu SC, Xu Y, Balschi JA and Springer CS Jr: Bulk magnetic susceptibility shifts in NMR studies of compartmentalized samples: use of paramagnetic reagents. Magn Reson Med. 13:239–262. 1990. View Article : Google Scholar : PubMed/NCBI
71	Kayar SR, Hoppeler H, Howald H, Claassen H and Oberholzer F: Acute effects of endurance exercise on mitochondrial distribution and skeletal muscle morphology. Eur J Appl Physiol Occup Physiol. 54:578–584. 1986. View Article : Google Scholar : PubMed/NCBI
72	Canavoso LE, Jouni ZE, Karnas KJ, Pennington JE and Wells MA: Fat metabolism in insects. Annu Rev Nutr. 21:23–46. 2001. View Article : Google Scholar
73	Gilby AR: Lipids and their metabolism in insects. Annu Rev Entomol. 10:141–160. 1965. View Article : Google Scholar
74	Fast PG: A comparative study of the phospholipids and fatty acids of some insect lipids. Science. 155:1680–1681. 1967.
75	Stanley-Samuelson DW, Jurenka RA, Cripps C, Blomquist GJ and deRenobales M: Fatty acids in insects: composition, metabolism, and biological significance. Arch Insect Biochem Physiol. 9:1–33. 1988. View Article : Google Scholar
76	Horne I, Haritos VS and Oakeshott JG: Comparative and functional genomics of lipases in holometabolous insects. Insect Biochem Mol Biol. 39:547–567. 2009. View Article : Google Scholar : PubMed/NCBI
77	Patel RT, Soulages JL, Hariharasundaram B and Arrese EL: Activation of the lipid droplet controls the rate of lipolysis of triglycerides in the insect fat body. J Biol Chem. 280:22624–22631. 2005. View Article : Google Scholar : PubMed/NCBI
78	Bharucha KN, Tarr P and Zipursky SL: A glucagon-like endocrine pathway in Drosophila modulates both lipid and carbohydrate homeostasis. J Exp Biol. 211:3103–3110. 2008. View Article : Google Scholar : PubMed/NCBI
79	Righi V, Apidianakis Y, Mintzopoulos D, Astrakas L, Rahme LG and Tzika AA: In vivo high-resolution magic angle spinning magnetic resonance spectroscopy of Drosophila melanogaster at 14.1 T shows trauma in aging and in innate immune-deficiency is linked to reduced insulin signaling. Int J Mol Med. 26:175–184. 2010.
80	Machann J, Thamer C, Schnoedt B, et al: Age and gender related effects on adipose tissue compartments of subjects with increased risk for type 2 diabetes: a whole body MRI/MRS study. MAGMA. 18:128–137. 2005. View Article : Google Scholar : PubMed/NCBI
81	Nakagawa Y, Hattori M, Harada K, Shirase R, Bando M and Okano G: Age-related changes in intramyocellular lipid in humans by in vivo H-MR spectroscopy. Gerontology. 53:218–223. 2007. View Article : Google Scholar : PubMed/NCBI
82	Muller MJ and Herndon DN: The challenge of burns. Lancet. 343:216–220. 1994. View Article : Google Scholar : PubMed/NCBI
83	Ikezu T, Okamoto T, Yonezawa K, Tompkins RG and Martyn JA: Analysis of thermal injury-induced insulin resistance in rodents. Implication of postreceptor mechanisms. J Biol Chem. 272:25289–25295. 1997. View Article : Google Scholar : PubMed/NCBI
84	Sinha R, Dufour S, Petersen KF, et al: Assessment of skeletal muscle triglyceride content by (1)H nuclear magnetic resonance spectroscopy in lean and obese adolescents: relationships to insulin sensitivity, total body fat, and central adiposity. Diabetes. 51:1022–1027. 2002. View Article : Google Scholar
85	Schrauwen-Hinderling VB, Hesselink MK, Schrauwen P and Kooi ME: Intramyocellular lipid content in human skeletal muscle. Obesity (Silver Spring). 14:357–367. 2006. View Article : Google Scholar : PubMed/NCBI
86	Consitt LA, Bell JA and Houmard JA: Intramuscular lipid metabolism, insulin action, and obesity. IUBMB Life. 61:47–55. 2009. View Article : Google Scholar : PubMed/NCBI
87	Johnson AB, Argyraki M, Thow JC, Cooper BG, Fulcher G and Taylor R: Effect of increased free fatty acid supply on glucose metabolism and skeletal muscle glycogen synthase activity in normal man. Clin Sci (Lond). 82:219–226. 1992.PubMed/NCBI
88	Clancy DJ, Gems D, Harshman LG, et al: Extension of life-span by loss of CHICO, a Drosophila insulin receptor substrate protein. Science. 292:104–106. 2001. View Article : Google Scholar : PubMed/NCBI
89	Bohni R, Riesgo-Escovar J, Oldham S, et al: Autonomous control of cell and organ size by CHICO, a Drosophila homolog of vertebrate IRS1–4. Cell. 97:865–875. 1999. View Article : Google Scholar : PubMed/NCBI
90	Paumen MB, Ishida Y, Muramatsu M, Yamamoto M and Honjo T: Inhibition of carnitine palmitoyltransferase I augments sphingolipid synthesis and palmitate-induced apoptosis. J Biol Chem. 272:3324–3329. 1997. View Article : Google Scholar : PubMed/NCBI
91	Ruddock MW, Stein A, Landaker E, et al: Saturated fatty acids inhibit hepatic insulin action by modulating insulin receptor expression and post-receptor signalling. J Biochem. 144:599–607. 2008. View Article : Google Scholar : PubMed/NCBI
92	Fernandez-Real JM and Pickup JC: Innate immunity, insulin resistance and type 2 diabetes. Trends Endocrinol Metab. 19:10–16. 2008. View Article : Google Scholar