Direct visualization of CHIP-mediated degradation of alpha-synuclein in vivo: implications for PD therapeutics.

Parkinson's disease is a neurodegenerative disorder characterized by Lewy bodies, a pathological hallmark comprised mostly of aggregated alpha synuclein. Accumulating evidence demonstrates the association of smaller oligomeric aggregates to disease etiology and many therapeutic approaches are aimed at inhibiting and reducing the aggregation process. Molecular chaperones and co-chaperones play a key role in protein homeostasis and have potential as therapeutics to inhibit alpha synuclein associated toxicity. Here we use a gene therapy approach to evaluate the applicability of the Hsp70 co-chaperone CHIP (C-terminal Hsp70 interacting protein) as a therapeutic candidate and examine its direct effect on alpha synuclein aggregates in vivo. Utilizing a novel viral vector mediated rat model to directly detect alpha synuclein aggregates, we show that CHIP can mediate the degradation of alpha synuclein aggregates in vivo. However, our studies also reveal that CHIP may potentially degrade tyrosine hydroxylase which would compromise the applicability of CHIP as a therapeutic approach for Parkinson's disease.

Abnormal involuntary movement (AIM) expression following D2 dopamine agonist challenge is determined by the nature of prior dopamine receptor stimulation (priming) in 6-hydroxydopamine lesioned rats.

Rats with unilateral 6-hydroxydopamine (6-OHDA) lesions show sensitization (priming) of rotational behavior upon repeated treatment with dopamine agonists. To relate these observations to dyskinesias exhibited by Parkinson's Disease patients, we assessed abnormal involuntary movements (AIMs) in 6-OHDA rats, which were primed with three injections of either the following: water, D1/D2 agonist apomorphine (Apo) (0.5mg/kg), D1 agonist SKF38393 (SKF) (10mg/kg) or D2 agonist quinpirole (Quin) (1 or 2.5mg/kg). The rats were challenged one week later with Quin (0.25mg/kg). Axial, limb, orolingual, locomotor, and grooming AIMs were scored (0-4) every 5min. Priming with water did not produce AIMs. Priming with Quin (1mg/kg) produced axial and locomotor AIMs, while priming with Apo, SKF or Quin (2.5mg/kg) produced axial, locomotor, limb, and grooming AIMs. The disparity in AIM profiles between Quin (1mg/kg) and (2.5mg/kg) was not the result of D1 receptor stimulation since there was little striatal Fos expression following the third priming injection with Quin (1 or 2.5mg/kg) compared to following SKF, which led to robust striatal Fos expression. Challenge with Quin (0.25mg/kg) essentially reproduced the categories of AIMs exhibited during priming, with no AIMs in water-primed 6-OHDA rats, mild, non-significant, axial and locomotor AIMs in Quin (1 and 2.5mg/kg)-primed 6-OHDA rats, and axial, limb, locomotor, and grooming AIMs in Apo- and SKF-primed 6-OHDA rats. These data suggest that the types of AIMs expressed following challenge with Quin depend on the dopamine receptor subtype and dose of dopamine agonist used during priming.