Analysis of lysosomal hydrolase trafficking and activity in human iPSC-derived neuronal models

Leah K. Cuddy*, Joseph R. Mazzulli

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Lysosomes are critical for maintaining protein homeostasis and cellular metabolism. Lysosomal dysfunction and disrupted protein trafficking contribute to cell death in neurodegenerative disorders, including Parkinson's disease and dementia. We describe three complementary protocols—the use of protein glycosylation, western blotting, immunofluorescence, and hydrolase activity measurement—to analyze the trafficking and activity of lysosomal proteins in patient-derived neurons differentiated from iPSCs. These methods should help to identify lysosomal phenotypes in patient-derived cultures and aid the discovery of therapeutics that augment lysosomal function. For complete details on the use and execution of this protocol, please refer to Cuddy et al. (2019).

Original languageEnglish (US)
Article number100340
JournalSTAR Protocols
Volume2
Issue number1
DOIs
StatePublished - Mar 19 2021

Keywords

  • Neuroscience
  • Protein biochemistry
  • Stem cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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