BackgroundParkinson’s disease (PD) is characterized by a chronic loss of dopaminergic neurons and the presence of proteinaceous inclusions (Lewy bodies) within some remaining neurons in the substantia nigra. Recently, astroglial inclusion body has also been found in some neurodegenerative diseases including PD. However, the underlying molecular mechanisms of how astroglial protein aggregation forms remain largely unknown. Here, we investigated the contribution of αB-crystallin (CRYAB), a small heat shock protein, in α-synuclein inclusion formation in astrocytes.MethodsSmall interfering RNA (siRNA)-mediated CRYAB (siCRYAB) knockdown or CRYAB overexpression was performed to investigate the impact of CRYAB on the autophagy in human glioblastoma cell line U251 cells. Co-immunoprecipitation (co-IP) and immunoblotting were used to dissect the interaction among multiple proteins. The clearance of α-synuclein in vitro was evaluated by immunocytochemistry. CRYAB transgenic mice and transgenic mice overexpressing A30P mutant form of human α-synuclein were used to examine the influence of CRYAB to α-synuclein accumulation in vivo.ResultsWe found that knockdown of CRYAB in U251 cells or primary cultured astrocytes resulted in a marked augmentation of autophagy activity. In contrast, exogenous CRYAB disrupted the assembly of the BAG3-HSPB8-HSC70 complex via binding with BAG3, thereby suppressing the autophagy activity. Furthermore, CRYAB-regulated autophagy has relevance to PD pathogenesis. Knockdown of CRYAB remarkably promoted cytoplasmic clearance of α-synuclein preformed fibrils (PFFs). Conversely, selective overexpression of CRYAB in astrocytes markedly suppressed autophagy leading to the accumulation of α-synuclein aggregates in the brain of transgenic mice expressing human α-synuclein A30P mutant.ConclusionsThis study reveals a novel function for CRYAB as a natural inhibitor of astrocytic autophagy and shows that knockdown of CYRAB may provide a therapeutic target against proteinopathies such as synucleinopathies.

We have studied the multiphoton dissociation dynamics of the Ẽ Ј 1 A 1 Ј Rydberg state of ammonia ͑NH 3 ͒ on a homebuilt femtosecond pump-probe system by resonance-enhanced multiphoton ionization photoelectron ͑REMPI-PE͒ spectroscopy. The highly excited Rydberg state, Ẽ Ј 1 A 1 Ј, of ammonia was accessed by two 267 nm pump photons and then ionized by a 401 nm probe pulse delayed in time. The variation of the REMPI-PE spectra of ammonia with pump-probe delay time provides valuable information on the dynamics of the excited intermediate accessed by the pump pulse. We find that the Frank-Condon preferred transition during ionization does not occur for ⌬ 1 = 0 but for ⌬ 1 = 1, which implies that the intermediate has a different geometry from the ionic ground state. Different dynamical behavior has been observed for each of the transitions ⌬ 1 =0,1,2,3, giving a full temporal description of the excited intermediate state by projection onto the eigenspace of the ionic ground state.