FLUORESCENCE SPECTROSCOPY ON HUNTINGTON'S FIBROBLASTS

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Abstract
Abstract– 8‐Anilino‐1‐naphthalene sulfonic acid (ANS), a fluorescent probe specific for membrane polar‐apolar interfaces, has been used to investigate differences in corrected excitation, emission and scanned excitation polarization spectra of intact age, sex and passage number matched Huntington's disease (HD) and normal fibroblasts grown in cell culture. At passage number 4, the HD fibroblasts, compared to normals, exhibited a 30‐fold increase in excitation and emission intensity, a 30 nm blue shift of the emission maximum and a 0.18 polarization unit increase. These findings suggest that the ANS molecule experiences either increased binding sites or an increased quantum yield, a more nonpolar environment, and a more restricted motion in HD fibroblasts. Continued subculture at higher passages (10) showed a change of these findings towards normals. These fluorescent studies support the concept that a membrane abnormality is present in HD.