Axon-dominant localization of cell-surface cholesterol in cultured hippocampal neurons and its disappearance in Niemann-Pick type C model cells.

There is growing evidence showing the important role of cholesterol in maintaining neuronal function. In particular, much attention has been paid to the role of the cholesterol-rich microdomains called lipid rafts. However, the cholesterol distribution on neurons is not clear. Here, we investigated localization of cholesterol in cultured rat hippocampal neurons, using filipin and a novel cholesterol-binding reagent BCtheta. In our culture system, BCtheta detects only cell-surface cholesterol, whereas filipin stains both intracellular and cell-surface cholesterol. BCtheta staining appeared visible in a maturation-dependent manner and showed axon-dominant distribution of cell-surface cholesterol in fully matured neurons. A part of this cholesterol on axons was resistant to detergents at 4 degrees C, and thus might be involved in lipid rafts. Interestingly, Niemann-Pick type C model neurons induced by class 2 amphiphiles lost the cell-surface but not the intracellular cholesterol staining. Niemann-Pick type C disease is caused by the disruption of intracellular cholesterol transport and is known to induce neurodegeneration in brains accompanied by formation of neurofibrillary tangles. Our observations suggest the important role of cell-surface cholesterol in maintaining a functional axonal membrane and indicate that the observed defect in axonal surface cholesterol might lead to neurodegeneration.