α-Synuclein Seeding Assay Using Cultured Cells

Methods Mol Biol. 2021:2322:27-39. doi: 10.1007/978-1-0716-1495-2_3.

Abstract

α-Synuclein, a presynaptic protein, is involved in synaptic vesicle dynamics in response to neuronal activity. Mutations of the α-synuclein gene and the neuronal deposition of α-synuclein, called Lewy bodies, are linked to the development of Parkinson's disease. α-Synuclein has a prion-like property that converts its physiological protein conformation to a pathogenic one, forming disease-causing fibrils. Aggregation of these fibrils and subsequent inclusion formation are suggested to interfere with vesicular trafficking and organelle function in neurons. Thus, detection of a prion-like property of α-synuclein and the evaluation of its modifying factors are required to understand the pathogenesis of Parkinson's disease and to develop new therapies. In this chapter, we describe a cell-based assay for detecting α-synuclein propagation.

Keywords: Cultured cells; Protein purification; Recombinant protein; Transfection; α-Synuclein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport / physiology
  • Brain / metabolism
  • Cell Line, Tumor
  • Cells, Cultured / metabolism*
  • Humans
  • Lewy Bodies / metabolism
  • Neurons / metabolism
  • Parkinson Disease / metabolism
  • Prions / metabolism
  • alpha-Synuclein / metabolism*

Substances

  • Prions
  • alpha-Synuclein