Post-transcriptional Inhibition of Hsc70-4/HSPA8 Expression Leads to Synaptic Vesicle Cycling Defects in Multiple Models of ALS

Cell Rep. 2017 Oct 3;21(1):110-125. doi: 10.1016/j.celrep.2017.09.028.

Abstract

Amyotrophic lateral sclerosis (ALS) is a synaptopathy accompanied by the presence of cytoplasmic aggregates containing TDP-43, an RNA-binding protein linked to ∼97% of ALS cases. Using a Drosophila model of ALS, we show that TDP-43 overexpression (OE) in motor neurons results in decreased expression of the Hsc70-4 chaperone at the neuromuscular junction (NMJ). Mechanistically, mutant TDP-43 sequesters hsc70-4 mRNA and impairs its translation. Expression of the Hsc70-4 ortholog, HSPA8, is also reduced in primary motor neurons and NMJs of mice expressing mutant TDP-43. Electrophysiology, imaging, and genetic interaction experiments reveal TDP-43-dependent defects in synaptic vesicle endocytosis. These deficits can be partially restored by OE of Hsc70-4, cysteine-string protein (Csp), or dynamin. This suggests that TDP-43 toxicity results in part from impaired activity of the synaptic CSP/Hsc70 chaperone complex impacting dynamin function. Finally, Hsc70-4/HSPA8 expression is also post-transcriptionally reduced in fly and human induced pluripotent stem cell (iPSC) C9orf72 models, suggesting a common disease pathomechanism.

Keywords: C9orf72; Drosophila; RNA processing; TDP-43; amyotrophic lateral sclerosis; endocytosis; iPSC; neuromuscular junction; synaptic vesicle cycle; translation.

MeSH terms

  • Amyotrophic Lateral Sclerosis / genetics*
  • Amyotrophic Lateral Sclerosis / metabolism
  • Amyotrophic Lateral Sclerosis / pathology
  • Animals
  • C9orf72 Protein / genetics
  • C9orf72 Protein / metabolism
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Disease Models, Animal
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / metabolism
  • Dynamins / genetics
  • Dynamins / metabolism
  • Endocytosis
  • Gene Expression Regulation
  • HSC70 Heat-Shock Proteins / genetics*
  • HSC70 Heat-Shock Proteins / metabolism
  • HSP40 Heat-Shock Proteins / genetics
  • HSP40 Heat-Shock Proteins / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Motor Neurons / metabolism
  • Motor Neurons / pathology
  • Neuromuscular Junction / metabolism
  • Neuromuscular Junction / pathology
  • Protein Aggregates
  • Protein Biosynthesis
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Synaptic Transmission
  • Synaptic Vesicles / metabolism*
  • Synaptic Vesicles / pathology

Substances

  • C9orf72 Protein
  • C9orf72 protein, human
  • DNA-Binding Proteins
  • HSC70 Heat-Shock Proteins
  • HSP40 Heat-Shock Proteins
  • HSPA8 protein, human
  • Membrane Proteins
  • Protein Aggregates
  • RNA, Messenger
  • TARDBP protein, human
  • cysteine string protein
  • Dynamins