17-AAG induces cytoplasmic alpha-synuclein aggregate clearance by induction of autophagy.
BACKGROUND
The accumulation and aggregation of alpha-synuclein in nerve cells and glia are characteristic features of a number of neurodegenerative diseases termed synucleinopathies. alpha-Synuclein is a highly soluble protein which in a nucleation dependent process is capable of self-aggregation. The causes underlying aggregate formation are not yet understood, impairment of the proteolytic degradation systems might be involved.


RESULTS
In the present study the possible aggregate clearing effects of the geldanamycin analogue 17-AAG (17-(Allylamino)-17-demethoxygeldanamycin) was investigated. Towards this, an oligodendroglial cell line (OLN-93 cells), stably expressing human alpha-synuclein (A53T mutation) was used. In these cells small punctate aggregates, not staining with thioflavine S, representing prefibrillary aggregates, occur characteristically. Our data demonstrate that 17-AAG attenuated the formation of alpha-synuclein aggregates by stimulating macroautophagy. By blocking the lysosomal compartment with NH(4)Cl the aggregate clearing effects of 17-AAG were abolished and alpha-synuclein deposits were enlarged. Analysis of LC3-II immunoreactivity, which is an indicator of autophagosome formation, further revealed that 17-AAG led to the recruitment of LC3-II and to the formation of LC3 positive puncta. This effect was also observed in cultured oligodendrocytes derived from the brains of newborn rats. Inhibition of macroautophagy by 3-methyladenine prevented 17-AAG induced occurrence of LC3 positive puncta as well as the removal of alpha-synuclein aggregates in OLN-A53T cells.


CONCLUSIONS
Our data demonstrate for the first time that 17-AAG not only causes the upregulation of heat shock proteins, but also is an effective inducer of the autophagic pathway by which alpha-synuclein can be removed. Hence geldanamycin derivatives may provide a means to modulate autophagy in neural cells, thereby ameliorating pathogenic aggregate formation and protecting the cells during disease and aging.
Riedel
Michael
M
Department of Biology, Molecular Neurobiology, University of Oldenburg, Oldenburg, Germany.
Goldbaum
Olaf
O
Schwarz
Lisa
L
Schmitt
Sebastian
S
Richter-Landsberg
Christiane
C
D000818:Animals; D001343:Autophagy; D016227:Benzoquinones / Q000494:pharmacology*; D002460:Cell Line; D003593:Cytoplasm / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D047029:Lactams, Macrocyclic / Q000494:pharmacology*; D009154:Mutation; D051381:Rats; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism*
D016227:Benzoquinones; D047029:Lactams, Macrocyclic; D051844:alpha-Synuclein; C112765:tanespimycin