1. Hsp70 Gene Transfer by Adeno-associated Virus Inhibits MPTP-Induced Nigrostriatal Degeneration in the Mouse Model of Parkinson Disease 
Zhizhong Dong, David P. Wolfer, Hans-Peter Lipp, Hansruedi Büeler 
Molecular Therapy 
Volume 11, Issue 1, Pages (January 2005)
DOI: /j.ymthe
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

2. Fig. 1
Expression of EGFP and HA-Hsp70 in the substantia nigra of rAAV-injected mice. Expression of (A) EGFP and (D) HA-Hsp70 in tyrosine hydroxylase (TH)-positive neurons (B and E) of the substantia nigra pars compacta is shown. (C and F) Overlays. EGFP was detected in the FITC channel (without antibodies). HA-Hsp70 was detected by immunohistochemistry with an anti-HA antibody. No transgene expression was observed in the uninjected hemisphere or in the absence of the primary antibody (data not shown).
Molecular Therapy  , 80-88DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

3. Fig. 2
HA-Hsp70 inhibits MPTP-induced nigrostriatal degeneration in mice. Mice were injected above the right substantia nigra with the indicated rAAV vector and 5 weeks later treated with MPTP (chronic regimen). Animals were sacrificed and analyzed 9 days after the last MPTP injection. Normal control mice were age matched to the MPTP-treated mice. (A) Number of tyrosine hydroxylase (TH)-positive neurons in sections of the substantia nigra (mean ± SEM). Normal, 4 mice, n = 13 sections; AAV-EGFP, 5 mice, n = 16 sections; AAV-Hsp70, 12 mice, n = 37 sections. (B) Striatal dopamine levels expressed in ng/mg wet tissue (mean ± SEM). Normal, n = 6 mice; AAV-EGFP, n = 11 mice; AAV-Hsp70, n = 12 mice. (C) Percentage TH fiber density (mean ± SD). Normal, 4 mice, n = 14 sections; AAV-EGFP, 6 mice, n = 18 sections; AAV-Hsp70, 7 mice, n = 23 sections. (D–F) Pictures of DAB-stained TH-positive neurons of a (D) normal, (E) MPTP-treated, AAV-EGFP-injected, and (F) MPTP-treated, AAV-Hsp70-injected substantia nigra. (G) Illustration of TH fiber protection in the virus-injected striatal hemisphere of an AAV-Hsp70-injected mouse (stained with an antibody to TH).
Molecular Therapy  , 80-88DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

4. Fig. 3
Amphetamine-induced behavior. Behavioral analysis showing box plots of amphetamine-induced on-spot rotations and general turning bias during locomotion. Boxes indicate the 25th and 75th, whiskers the 10th and 90th percentiles. AAV-Hsp70 and AAV-EGFP groups were compared using the Mann–Whitney U test. The injection of amphetamine induced marked locomotion in all animals, interrupted by short rests and episodes of lingering. By direct observation during the experiment, on-spot rotations were noted in only one animal of the Hsp70 group, which earned a score of 2.08 rounds per minute (RPM) (black dot). Quantitative analysis of on-spot rotations was in agreement with direct observation and revealed no significant group difference (ns). Evaluation of general turning bias during locomotion showed a significant bias toward left turns in the AAV-Hsp70 group, while the AAV-EGFP group showed no turning bias. Overall locomotor activity was slightly higher in the AAV-Hsp70 group but this did not reach statistical significance (data not shown).
Molecular Therapy  , 80-88DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

5. Fig. 4
Hsp70 inhibits MPTP-induced apoptosis. Mice were sacrificed and analyzed 9 days after the last MPTP injection. (A) TUNEL-stained apoptotic nuclei in the substantia nigra of MPTP-treated mice. Sections from normal mice treated with DNase I served as a positive control. (B) Quantitation of apoptotic nuclei (mean ± SEM) in the substantia nigra (identified by the presence of the MTN). Normal, 3 mice, n = 9 sections; AAV-EGFP, 4 mice, n = 12 sections; AAV-Hsp70, 12 mice, n = 36 sections.
Molecular Therapy  , 80-88DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions