Singer H S, Tiemeyer M, Slesinger P A, Sinnott M L
Ann Neurol. 1987 May;21(5):497-503. doi: 10.1002/ana.410210514.
This study was designed to establish an in vitro model with biochemical and morphological similarities to the human neurodegenerative disease GM1 gangliosidosis. Utilizing a specific inactivator of the lysosomal enzyme GM1-ganglioside beta-galactosidase (beta-D-galactopyranosylmethyl-p-nitrophenyltriazene [beta-GalMNT]) and neuroblastoma X glioma hybrid cells (NG108-15), we suppressed beta-galactosidase activity for up to 72 hours. Coincidental with suppression of this enzyme to levels less than 1% of control, we found up to a nine-fold accumulation of its substrate, the GM1-ganglioside, and the ultrastructural appearance of membranous cytoplasmic bodies. beta-GalMNT treatment suppressed growth but had little effect on the specific activity of choline acetyltransferase, lactate dehydrogenase, or other lysosomal enzymes including galactosylceramidase. This model should permit studies of the neurophysiological effects of increased ganglioside accumulation and their reversibility.
本研究旨在建立一种体外模型,使其在生化和形态学上与人类神经退行性疾病GM1神经节苷脂贮积症相似。利用溶酶体酶GM1-神经节苷脂β-半乳糖苷酶的特异性灭活剂(β-D-吡喃半乳糖基甲基对硝基苯基三氮烯[β-GalMNT])和神经母细胞瘤X胶质瘤杂交细胞(NG108-15),我们将β-半乳糖苷酶活性抑制长达72小时。在将该酶抑制至低于对照水平1%的同时,我们发现其底物GM1-神经节苷脂积累高达九倍,并出现了膜性细胞质体的超微结构外观。β-GalMNT处理抑制了生长,但对胆碱乙酰转移酶、乳酸脱氢酶或其他溶酶体酶(包括半乳糖基神经酰胺酶)的比活性影响很小。该模型应有助于研究神经节苷脂积累增加的神经生理效应及其可逆性。