First Author | Arthur JR | Year | 2011 |
Journal | J Lipid Res | Volume | 52 |
Issue | 7 | Pages | 1345-51 |
PubMed ID | 21508255 | Mgi Jnum | J:174698 |
Mgi Id | MGI:5140641 | Doi | 10.1194/jlr.M012633 |
Citation | Arthur JR, et al. (2011) Filipin recognizes both GM1 and cholesterol in GM1 gangliosidosis mouse brain. J Lipid Res 52(7):1345-51 |
abstractText | Filipin is an antibiotic polyene widely used as a histochemical marker for cholesterol. We previously reported cholesterol/filipin-positive staining in brain of beta-galactosidase (beta-gal) knockout ((-/-)) mice (GM1 gangliosidosis). The content and distribution of cholesterol and gangliosides was analyzed in plasma membrane (PM) and microsomal (MS) fractions from whole-brain tissue of 15 week-old control (beta-gal(+/-)) and GM1 gangliosidosis (beta-gal(-/-)) mice. Total ganglioside content (mug sialic acid/mg protein) was 3-fold and 7-fold greater in the PM and MS fractions, respectively, in betagal(-/-) mice than in betagal(+/-) mice. GM1 content was 30-fold and 50-fold greater in the PM and MS fractions, respectively. In contrast, unesterified cholesterol content (mug/mg protein) was similar in the PM and the MS fractions of the betagal(-/-) and betagal(+/-) mice. Filipin is known to bind to various sterol derivatives and phospholipids on thin-layer chromatograms. Biochemical evidence is presented showing that filipin also binds to GM1 with an affinity similar to that for cholesterol, with a corresponding fluorescent reaction. Our data suggest that the GM1 storage seen in the beta-gal(-/-) mouse contributes to the filipin ultraviolet fluorescence observed in GM1 gangliosidosis brain. The data indicate that in addition to cholesterol, filipin can also be useful for detecting GM1. |