Ser129 phosphorylation of endogenous α-synuclein induced by overexpression of polo-like kinases 2 and 3 in nigral dopamine neurons is not detrimental to their survival and function

Neurobiol Dis. 2015 Jun:78:100-14. doi: 10.1016/j.nbd.2015.03.008. Epub 2015 Mar 25.

Abstract

Phosphorylation of the α-synuclein (α-syn) protein at Ser129 [P(S129)-α-syn] was found to be the most abundant form in intracellular inclusions in brains from Parkinson's disease (PD) patients. This finding suggests that P(S129)-α-syn plays a central role in the pathogenesis of PD. However, it is at present unclear whether P(S129)-α-syn is pathogenic driving the neurodegenerative process. Rodent studies using neither the phosphomimics of human α-syn nor co-expression of human wild-type α-syn and kinases phosphorylating α-syn at Ser129 gave consistent results. One major concern in interpreting these findings is that human α-syn was expressed above physiological levels inducing neurodegeneration in rat nigral neurons. In order to exclude this confounding factor, we took a different approach and increased the phosphorylation level of endogenous α-syn. For this purpose, we took advantage of recombinant adeno-associated viral (rAAV) vectors to deliver polo-like kinase (PLK) 2 or PLK3 in the substantia nigra and investigated whether increased levels of P(S129)-α-syn compromised the function and survival of nigral dopaminergic neurons. Interestingly, we observed that hyperphosphorylated α-syn did not induce nigral dopaminergic cell death, as assessed at 1 and 4months. Furthermore, histological analysis did not show any accumulation of α-syn protein or formation of inclusions. Using in vivo microdialysis, we found that the only measurable functional alteration was the depolarisation-induced release of dopamine, while the in vivo synthesis rate of DOPA and dopamine baseline release remained unaltered. Taken together, our results suggest that phosphorylation of α-syn at Ser129 does not confer a toxic gain of function per se.

Keywords: Adeno-associated vector; Parkinson's disease; Phosphorylation; Polo-like kinase; α-Synuclein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism
  • Brain / virology
  • Cell Cycle Proteins / metabolism*
  • Cell Survival
  • Dependovirus
  • Dopamine / metabolism
  • Dopaminergic Neurons / metabolism*
  • Female
  • Genetic Vectors / adverse effects
  • Phosphorylation
  • Protein Serine-Threonine Kinases / metabolism*
  • Rats
  • Rats, Sprague-Dawley
  • Serine / metabolism
  • Substantia Nigra / metabolism*
  • Substantia Nigra / virology
  • alpha-Synuclein / metabolism*

Substances

  • Cell Cycle Proteins
  • alpha-Synuclein
  • Serine
  • Plk3 protein, rat
  • Protein Serine-Threonine Kinases
  • Plk2 protein, rat
  • Dopamine