High-throughput imaging method for direct assessment of GM1 ganglioside levels in mammalian cells

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• 作者：Walter Acosta^a ; ^{wacosta@biostrategies-lc.com" class="auth_mail" title="E-mail the corresponding author} ; Reid Martin^a ; ^b ; David N. Radin^a ; Carole L. Cramer^a ; ^b
• 关键词：High-throughput imaging ; GM1-gangliosidosis ; Acid β-galactosidase ; Cholera toxin B subunit
• 刊名：Data in Brief
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：6
• 期：Complete
• 页码：1016-1022
• 全文大小：1506 K

文摘

GM1-gangliosidosis is an inherited autosomal recessive disorder caused by mutations in the gene GLB1, which encodes acid β-galactosidase (β-gal). The lack of activity in this lysosomal enzyme leads to accumulation of GM1 gangliosides (GM1) in cells. We have developed a high-content-imaging method to assess GM1 levels in fibroblasts that can be used to evaluate substrate reduction in treated GLB1^−/− cells [1]. This assay allows fluorescent quantification in a multi-well system which generates unbiased and statistically significant data. Fluorescently labeled Cholera Toxin B subunit (CTXB), which specifically binds to GM1 gangliosides, was used to detect in situ GM1 levels in a fixed monolayer of fibroblasts. This sensitive, rapid, and inexpensive method facilitates in vitro drug screening in a format that allows a high number of replicates using low working volumes.