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Evidence That Mobile Lipids Detected in Rat Brain Glioma by ¹H Nuclear Magnetic Resonance Correspond to Lipid Droplets¹

INSERM U438, RMN Bioclinique, CHU de Grenoble, BP 217, 38043 Grenoble Cedex 9, France [C. R., N. F., E. S-L., H. L., M. I., A. Z., R. M., M. D.], and Universitat Autonòma de Barcelona, Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, 08193 Bellaterra, Spain [I. B., M-G. C., A. M., C. A.]

Mobile lipids have been detected by proton nuclear magnetic resonance (NMR) in animal and human tumors (cultured cells, biopsies, and in vivo), but their origin and subcellular location are still unclear. They have been associated with malignancy, metastatic ability, drug resistance, and necrosis. We wanted to determine whether these lipids are located within plasma membrane microdomains or in lipid droplets for a C6 cell-induced rat glioma. NMR-visible mobile lipids were found in all subcellular fractions isolated from the rat tumor, except in the cytosolic supernatants. Transmission electron microscopy showed that lipid droplets were present in all subcellular fractions containing NMR-visible lipids and in the necrotic and perinecrotic areas of the tumor. The mean diameter of droplets isolated by flotation in the subcellular fractionation protocol was 0.97 µm (n = 682; droplet profile diameter range between 0.2 and 5.0 µm). The apparent diffusion coefficient for these lipids (46 ± 17 µm²s^-1) measured in vivo by proton spectroscopy was four orders of magnitude higher than would be expected if mobile lipids were inside plasma membrane microdomains. The combined results demonstrated that mobile lipids detected in vivo by proton NMR in the C6 rat glioma are located in large lipid droplets, associated with the necrotic process.

¹ This work was supported by the Association Espoir, l'Isère contre le Cancer, Fondation pour la Recherche Médicale, and by Comisión Interministerial de Ciencia y Tecnología (CICYT) SAF 93-0582-C02-01. Short-term travel grants were obtained from Coopération franco-espagnole-Accords INSERM-CSIC. N. F. received a grant from the Ligue Nationale Contre le Cancer, and E. S-L. received a grant from the Association pour la Recherche sur le Cancer and Ligue Nationale Contre le Cancer. A. M. held a Fellowship from the Asociación Española Contra el Cancer during his participation in the study.

² To whom requests for reprints should be addressed.

Received 8/ 5/96. Accepted 12/ 4/96.

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