Abstract
Object
To evaluate how spinning rate affects mobile lipid (ML) resonances visibility in HR-MAS spectra of C6 glioma cells and artificial oil bodies (AOB), as models of cytosolic lipid droplets.
Materials and methods
Using C6 cells and AOB of two different sizes, 780 ± 580 and 240 ± 293 nm, as models, we acquired HR-MAS pulse and acquire spectra at different spinning rates between 500 and 15,000 Hz, all at 37 °C.
Results
Sample spinning at 15,000 Hz increased by 2.3 and 4.6-fold with respect to 500 Hz spinning the area of resonances corresponding to ML at 0.88 and 1.28 ppm, respectively, for log phase C6 cells. Furthermore, postconfluent C6 cells displayed an increase of 2.5-fold at 0.88 ppm and 4.2-fold at 1.28 ppm. These changes were reversible upon low speed spinning. AOBs did show much lower ML area increases (1.4–1.5-fold) upon high-speed HR-MAS.
Conclusion
ML can be reversibly mobilized in C6 glioma cells by high-speed HR-MAS, partially unveiling the NMR “invisible” ML pool. A small part of the ML pool also shows reduced visibility in freely tumbling AOBs.
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Abbreviations
- AOB:
-
Artificial oil bodies
- ChCCp:
-
Choline containing compounds
- HR-MAS:
-
High resolution-magic angle spinning
- LB:
-
Lysogeny broth
- LD:
-
Lipid droplets
- ML:
-
Mobile lipids
- PBS:
-
Phosphate buffered saline
- PtdCho:
-
Phosphatidylcholine
- TAG:
-
Triacylglycerides
- TLC:
-
Thin layer chromatography
- TMS:
-
Tetramethylsilane
References
Hakumäki JM, Kauppinen RA (2000) 1H NMR visible lipids in the life and death of cells. Trends Biochem Sci 25:357–362
Chen JH, Enloe BM, Weybright P, Campbell N, Dorfman D, Fletcher CD, Cory DG, Singer S (2002) Biochemical correlates of thiazolidinedione-induced adipocyte differentiation by high-resolution magic angle spinning NMR spectroscopy. Magn Reson Med 48(4):10–602
Barba I, Cabañas ME, Arús C (1999) The relationship between nuclear magnetic resonance-visible lipids, lipid droplets, and cell proliferation in cultured C6 cells. Cancer Res 59:1861–1868
Mountford CE, Wright LC (1988) Organization of lipids in the plasma membranes of malignant and stimulated cells: a new model. Trends Biochem Sci 13:172–177
Rémy C, Fouilhe N, Barba I, Sam-Lai E, Lahrech H, Cucurella MG, Izquierdo M, Moreno A, Ziegler A, Massarelli R, Décorps M, Arús C (1997) Evidence that mobile lipids detected in rat brain glioma by 1H nuclear magnetic resonance correspond to lipid droplets. Cancer Res 57:407–414
Quintero M, Cabañas ME, Arús C (2007) A possible cellular explanation for the NMR-visible mobile lipid (ML) changes in cultured C6 glioma cells with growth. Biochim Biophys Acta 177:31–44
Delikatny EJ, Chawla S, Leung DJ, Poptani H (2011) MR-visible lipids and the tumor microenviroment. NMR Biomed 24(6):592–611
Zweytick D, Athesntaedt K, Daum G (2000) Intracellular lipid particles of eukaryotic cells. Biochim Biophys Acta 1469:101–120
Martin S, Parton RG (2006) Lipid droplets: a unified view of a dynamic organelle. Nat Rev Mol Cell Bio 7:373–378
Pérez Y, Lahrech H, Cabañas ME, Barnadas R, Sabés M, Rémy C, Arús C (2002) Measurement by nuclear magnetic resonance diffusion of the dimensions of the mobile lipid compartment in C6 cells. Cancer Res 62:5672–5677
Beckonert O, Coen M, Keun HC, Wang Y, Ebbels TMD, Holmes E, Lindon JC, Nicholson JK (2010) High-resolution magic-angle-spinning NMR spectroscopy for metabolic profiling of intact tissues. Nat Protoc 5:1019–1032
Lindon JC, Beckonert OP, Holmes E, Nicholson JK (2009) High-resolution magic angle spinning NMR spectroscopy: application to biomedical studies. Prog Nucl Mag Res Sp 55(2):79–100
Martín-Sitjar J, Delgado-Goñi T, Valverde D, Cabañas, M, Nolis, P, Arús C (2008) Unveiling the “invisible” neutral lipid pool in cultured log phase C6 cells using HR-MAS at 37 °C. In: Proceedings of the 24th annual meeting, European Society for Magnetic Resonance in Medicine and Biology, pp 181–182
Traikia M, Langlais DB, Cannarozzi GM, Devaux PF (1997) High-Resolution spectra of liposomes using MAS NMR. The case of intermediate-size vesicles. J Magn Reson 125:140–144
Chen MCM, Chyan CL, Lee TTT, Huang SH, Tzen JTC (2004) Constitution of stable artificial oil bodies with triacylglycerol, phospholipids and caleosin. J Agric Food Chem 52:3982–3987
Tai SSK, Chen MCM, Peng CC, Tzen JTC (2002) Gene family of oleosin isoforms and their structural stabilization in sesame seed oil bodies. Biosci Biotechnol Biochem 66(10):2146–2153
Peng CC, Lin IP, Lin CK, Tzen JTC (2003) Size and stability of reconstituted sesame oil bodies. Biotechnol Progr 19(5):1623–1626
Tzen JTC, Huang AHC (1992) Surface structure and properties of plant seed oil bodies. J Cell Biol 117(2):327–335
Peng CC, Chen JCF, Shyu DJH, Chen MJ, Tzen JTC (2004) A system for purification of recombinant proteins in Escherichia coli via artificial oil bodies constituted with their oleosin-fused polypeptides. J Biotechnol 111:51–57
Liu TH, Chyan CL, Li FY, Tzen JT (2009) Stability of artificial oil bodies constituted with recombinant caleosins. J Agric Food Chem 57:2308–2313
Vuorela T, Catte A, Niemelä PS, Hall A, Hyvónen MT, Marrink SJ, Kattunen M, Vattulainen I (2010) Role of lipids in spheroidal high density lipoproteins. PLoS Comp Biol 6(10):e1000964
Hevonoja T, Pentikäinen MO, Hyvönen MT, Kovanen PT, Ala-Korpela M (2000) Structure of low density lipoprotein (LDL) particles: basis for understanding molecular changes in modified LDL. Biochem Biophys Acta 1488(3):189–210
Valverde-Saubí D, Candiota AP, Molins MA, Feliz M, Godino O, Dávila M, Acebes JJ, Arús C (2010) Short-term temperature effect on the HRMAS spectra of human brain tumor biopsies and their pattern recognition analysis. Magn Reson Mater Phy 23(4):203–215
Opstad KS, Bell BA, Griffiths JR, Howe FA (2008) An assessment of the effects of sample ischaemia and spinning time on the metabolic profile of brain tumour biopsy specimens as determined by high-resolution magic angle spinning 1H NMR. NMR Biomed 21:1138–1147
Deese AJ, Dratz EA, Hymel L, Fleischer S (1982) Proton NMR T1, T2, and T1 rho relaxation studies of native and reconstituted sarcoplasmic reticulum and phospholipid vesicles. Biophys J 37(1):16–207
Shapiro YE (1988) The paramagnetic hydrophilic probing of colloidal systems by NMR spectroscopy. Russ Chem Rev 57:1253–1272
Delikatny EJ, Cooper WA, Brammah S, Sathasivam N, Rideout DC (2002) Nuclear magnetic resonance-visible lipids induced by cationic lipophilic chemotherapeutic agents are accompanied by increased lipid droplet formation and damaged mitochondria. Cancer Res 62(5):1394–1400
Griffin JL, Lehtimäki KK, Valonen PK, Gröhn OHJ, Kettunen MI, Ylä-Herttuala S, Pitkänen A, Nicholson JK, Kauppinen RA (2003) Assignment of 1H nuclear magnetic resonance visible polyunsaturated fatty acids in BT4C gliomas undergoing ganciclovir-thymidine kinase gene therapy-induced programmed cell death. Cancer Res 63:3195–3201
Acknowledgments
Juana Martín was the recipient of a PhD fellowship from the Ministerio de Ciencia e Innovación (Spain) associated to project SAF2008-03323. We thank the Summer Program in Taiwan for contributing to the J.M. visit at the National Chung Hsing University group of Prof Jason Tzen. This work was supported by the Spanish Ministerio de Ciencia e Innovación (SAF2008-03323 and SAF2011-23870). CIBER-BBN is an initiative of Instituto de Salud Carlos III, Spain, which co-funded with EU-funds.
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Martín-Sitjar, J., Delgado-Goñi, T., Cabañas, M.E. et al. Influence of the spinning rate in the HR-MAS pattern of mobile lipids in C6 glioma cells and in artificial oil bodies. Magn Reson Mater Phy 25, 487–496 (2012). https://doi.org/10.1007/s10334-012-0327-6
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DOI: https://doi.org/10.1007/s10334-012-0327-6