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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.

Righi V, Apidianakis Y, Mintzopoulos D, Astrakas L, Rahme LG, Tzika AA - Int. J. Mol. Med. (2010)

Bottom Line: We demonstrated, for the first time, the feasibility of using HRMAS MRS for molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe.We showed that the metabolic HRMAS MRS profiles of injured, aged wild-type (wt) flies and of immune deficient (imd) flies were more similar to chico flies mutated at the chico gene in the insulin signaling pathway, which is analogous to insulin receptor substrate1-4 (IRS1-4) in mammals and less to those of adipokinetic hormone receptor (akhr) mutant flies, which have an obese phenotype.Our approach advances the development of novel in vivo non-destructive research approaches in Drosophila, suggests biomarkers for investigation of biomedical paradigms, and thus may contribute to novel therapeutic development.

View Article: PubMed Central - PubMed

Affiliation: NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital and Shriners Burn Institute, Harvard Medical School, Boston, MA 02114, USA.

ABSTRACT
In vivo magnetic resonance spectroscopy (MRS), a non-destructive biochemical tool for investigating live organisms, has yet to be used in the fruit fly Drosophila melanogaster, a useful model organism for investigating genetics and physiology. We developed and implemented a high-resolution magic-angle-spinning (HRMAS) MRS method to investigate live Drosophila at 14.1 T. We demonstrated, for the first time, the feasibility of using HRMAS MRS for molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe. We showed that the metabolic HRMAS MRS profiles of injured, aged wild-type (wt) flies and of immune deficient (imd) flies were more similar to chico flies mutated at the chico gene in the insulin signaling pathway, which is analogous to insulin receptor substrate1-4 (IRS1-4) in mammals and less to those of adipokinetic hormone receptor (akhr) mutant flies, which have an obese phenotype. We thus provide evidence for the hypothesis that trauma in aging and in innate immune-deficiency is linked to insulin signaling. This link may explain the mitochondrial dysfunction that accompanies insulin resistance and muscle wasting that occurs in trauma, aging and immune system deficiencies, leading to higher susceptibility to infection. Our approach advances the development of novel in vivo non-destructive research approaches in Drosophila, suggests biomarkers for investigation of biomedical paradigms, and thus may contribute to novel therapeutic development.

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Representative 2D 1H-1H TOBSY HRMAS spectrum of live Drosophila at 14.1 T. Small metabolites and lipid components were identified. Metabolites: alanine (Ala), ß-alanine (ß-Ala), arginine (Arg), glutamine (Gln), glutamate (Glu), phosphocholine (PC), phosphoethanolamine (PE), Taurine (Tau), α-glucose (α-Glc) and glycerol. Lipids components: CH3 (0.89 ppm), (CH2)n (1.33 ppm), CH2C-CO (1.58 ppm), CH2C═C (2.02 ppm), CH2C═O (2.24 ppm), CH═CH (5.33 ppm).
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f3-ijmm-26-02-0175: Representative 2D 1H-1H TOBSY HRMAS spectrum of live Drosophila at 14.1 T. Small metabolites and lipid components were identified. Metabolites: alanine (Ala), ß-alanine (ß-Ala), arginine (Arg), glutamine (Gln), glutamate (Glu), phosphocholine (PC), phosphoethanolamine (PE), Taurine (Tau), α-glucose (α-Glc) and glycerol. Lipids components: CH3 (0.89 ppm), (CH2)n (1.33 ppm), CH2C-CO (1.58 ppm), CH2C═C (2.02 ppm), CH2C═O (2.24 ppm), CH═CH (5.33 ppm).

Mentions: Metabolites that could not be assigned or were not visible using the 1D spectrum were detected using selected 2D experiments such as 2D TOBSY (Fig. 3), and HSQC (Fig. 4); and their assignment was confirmed by comparison with literature data. HSQC spectra revealed directly bonded carbon-proton pairs, thus enabling the assignment of singlets (which do not give correlations in homonuclear TOBSY spectra), and the discrimination among compounds having similar protons but diverse 13C chemical shifts. The experiments provided complete and unambiguous identification of the metabolic pattern characterizing Drosophila. The main mobile lipids and small metabolites are reported in Table II.


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.

Righi V, Apidianakis Y, Mintzopoulos D, Astrakas L, Rahme LG, Tzika AA - Int. J. Mol. Med. (2010)

Representative 2D 1H-1H TOBSY HRMAS spectrum of live Drosophila at 14.1 T. Small metabolites and lipid components were identified. Metabolites: alanine (Ala), ß-alanine (ß-Ala), arginine (Arg), glutamine (Gln), glutamate (Glu), phosphocholine (PC), phosphoethanolamine (PE), Taurine (Tau), α-glucose (α-Glc) and glycerol. Lipids components: CH3 (0.89 ppm), (CH2)n (1.33 ppm), CH2C-CO (1.58 ppm), CH2C═C (2.02 ppm), CH2C═O (2.24 ppm), CH═CH (5.33 ppm).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3722717&req=5

f3-ijmm-26-02-0175: Representative 2D 1H-1H TOBSY HRMAS spectrum of live Drosophila at 14.1 T. Small metabolites and lipid components were identified. Metabolites: alanine (Ala), ß-alanine (ß-Ala), arginine (Arg), glutamine (Gln), glutamate (Glu), phosphocholine (PC), phosphoethanolamine (PE), Taurine (Tau), α-glucose (α-Glc) and glycerol. Lipids components: CH3 (0.89 ppm), (CH2)n (1.33 ppm), CH2C-CO (1.58 ppm), CH2C═C (2.02 ppm), CH2C═O (2.24 ppm), CH═CH (5.33 ppm).
Mentions: Metabolites that could not be assigned or were not visible using the 1D spectrum were detected using selected 2D experiments such as 2D TOBSY (Fig. 3), and HSQC (Fig. 4); and their assignment was confirmed by comparison with literature data. HSQC spectra revealed directly bonded carbon-proton pairs, thus enabling the assignment of singlets (which do not give correlations in homonuclear TOBSY spectra), and the discrimination among compounds having similar protons but diverse 13C chemical shifts. The experiments provided complete and unambiguous identification of the metabolic pattern characterizing Drosophila. The main mobile lipids and small metabolites are reported in Table II.

Bottom Line: We demonstrated, for the first time, the feasibility of using HRMAS MRS for molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe.We showed that the metabolic HRMAS MRS profiles of injured, aged wild-type (wt) flies and of immune deficient (imd) flies were more similar to chico flies mutated at the chico gene in the insulin signaling pathway, which is analogous to insulin receptor substrate1-4 (IRS1-4) in mammals and less to those of adipokinetic hormone receptor (akhr) mutant flies, which have an obese phenotype.Our approach advances the development of novel in vivo non-destructive research approaches in Drosophila, suggests biomarkers for investigation of biomedical paradigms, and thus may contribute to novel therapeutic development.

View Article: PubMed Central - PubMed

Affiliation: NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital and Shriners Burn Institute, Harvard Medical School, Boston, MA 02114, USA.

ABSTRACT
In vivo magnetic resonance spectroscopy (MRS), a non-destructive biochemical tool for investigating live organisms, has yet to be used in the fruit fly Drosophila melanogaster, a useful model organism for investigating genetics and physiology. We developed and implemented a high-resolution magic-angle-spinning (HRMAS) MRS method to investigate live Drosophila at 14.1 T. We demonstrated, for the first time, the feasibility of using HRMAS MRS for molecular characterization of Drosophila with a conventional MR spectrometer equipped with an HRMAS probe. We showed that the metabolic HRMAS MRS profiles of injured, aged wild-type (wt) flies and of immune deficient (imd) flies were more similar to chico flies mutated at the chico gene in the insulin signaling pathway, which is analogous to insulin receptor substrate1-4 (IRS1-4) in mammals and less to those of adipokinetic hormone receptor (akhr) mutant flies, which have an obese phenotype. We thus provide evidence for the hypothesis that trauma in aging and in innate immune-deficiency is linked to insulin signaling. This link may explain the mitochondrial dysfunction that accompanies insulin resistance and muscle wasting that occurs in trauma, aging and immune system deficiencies, leading to higher susceptibility to infection. Our approach advances the development of novel in vivo non-destructive research approaches in Drosophila, suggests biomarkers for investigation of biomedical paradigms, and thus may contribute to novel therapeutic development.

Show MeSH
Related in: MedlinePlus