Direct evidence for ultrastructures of the α-synuclein-associated synaptic vesicle pool in presynaptic terminals

Biochim Biophys Acta Mol Basis Dis. 2024 Dec;1870(8):167494. doi: 10.1016/j.bbadis.2024.167494. Epub 2024 Sep 2.

Abstract

SNCA/PARK1 encodes α-synuclein, which is associated with familial Parkinson's disease. Despite its abundance in presynaptic terminals, the aggregation mechanism of α-synuclein and its relationship with Parkinson's disease have not yet been elucidated. Moreover, the ultrastructures of α-synuclein localization sites in neuronal presynaptic terminals remain unclear. Therefore, we herein generated transgenic mice expressing human α-synuclein tagged with mKate2 (hSNCA-mKate2 mice). These mice exhibited normal growth and fertility and had no motor dysfunction relative to their wild-type littermates, even at one year old. α-Synuclein-mKate2 accumulated in presynaptic terminals, particularly between Purkinje cells in the cerebellum and neurons in cerebellar nuclei. α-Synuclein-mKate2 was associated with the presynaptic marker, synaptophysin. In-resin CLEM and immunoelectron or electron microscopy revealed that α-synuclein-mKate2 localized on the surface of synaptic vesicles that were tightly arranged and assembled to form large synaptic pools in the cerebellum with negligible effects on the active zone. These results suggest that α-synuclein-associated ultrastructures in the presynaptic terminals of hSNCA-mKate2 mice reflect the structures of α-synuclein-assembled synaptic vesicle pools, and the size of vesicle pools increased. This transgenic mouse model will be a valuable tool for studying α-synuclein-associated synaptic vesicle pools.

Keywords: Synaptic vesicle pool; Transgenic mouse; α-Synuclein.

MeSH terms

  • Animals
  • Cerebellum / metabolism
  • Cerebellum / ultrastructure
  • Humans
  • Male
  • Mice
  • Mice, Transgenic*
  • Parkinson Disease / metabolism
  • Parkinson Disease / pathology
  • Presynaptic Terminals* / metabolism
  • Presynaptic Terminals* / ultrastructure
  • Purkinje Cells / metabolism
  • Purkinje Cells / ultrastructure
  • Synaptic Vesicles* / metabolism
  • Synaptic Vesicles* / ultrastructure
  • Synaptophysin / genetics
  • Synaptophysin / metabolism
  • alpha-Synuclein* / genetics
  • alpha-Synuclein* / metabolism

Substances

  • alpha-Synuclein
  • SNCA protein, human
  • Synaptophysin