1. Volume 10, Issue 1, Pages 67-75 (July 2004)
Prolonged biologically active transgene expression driven by HSV LAP2 in brain in vivo 
Veljko Puskovic, Darren Wolfe, James Goss, Shaohua Huang, Marina Mata, Joseph C. Glorioso, David J. Fink 
Molecular Therapy 
Volume 10, Issue 1, Pages (July 2004)
DOI: /j.ymthe
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

2. Fig. 1 Schematic of vector constructs.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

3. Fig. 2
Time line of the two animal studies. (A) 6-OHDA study. Five and a half months after intracranial inoculation of the vector rats received a single intrastriatal injection of 6-OHDA. At 6 months they were tested for amphetamine-induced rotation and sacrificed for histology. (B) MPTP study. After intracranial vector inoculation mice received MPTP daily (5 days of 7) for 13 weeks. Behavioral testing (paw reaching) was performed at weekly intervals; at 13 weeks the animals were sacrificed for histology.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

4. Fig. 3
Rats inoculated with QLGD showed a significant reduction in amphetamine-induced rotation 6 months after vector inoculation. The net number of ipsilateral rotations over 90 min is presented. Means ± SEM, *P < compared to Q0ZHG or QHGD.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

5. Fig. 4
Rats inoculated with QLGD showed a significant preservation of TH-IR neurons in the lesioned SN 6 months after vector inoculation. The right column (“injected”) shows the SN ipsilateral to the 6-OHDA injection into striatum, inoculated with vector Q0ZHG (top row), QHGD (second row), and QLGD (third row). Representative photomicrographs demonstrate the loss of TH-IR cells in QHGD-inoculated animals compared to the contralateral, unlesioned side. The ratio of number of neurons (ipsilateral/contralateral) was determined in counts of five or six sections per animal with four or five animals per group. Scale bar, 200 μm. Means ± SEM, *P < compared to Q0ZHG or QHGD.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

6. Fig. 5
Rats inoculated with QLGD showed a significant preservation of TH-IR and DAT-IR terminals in the striatum. Representative immunostained brain sections are shown. In each composite the striatum ipsilateral to 6-OHDA injection is indicated by an arrow, and sections from animals inoculated with Q0ZHG (top row), QHGD (second row), and QLGD (third row) are shown. The amount of immunoreactivity was quantitated using a PC-based image analysis system. The ratio was determined from five or six sections per animal with four or five animals per group. Means ± SEM, *P < compared to Q0ZHG or QLGD.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

7. Fig. 6
The amount of GDNF in substantia nigra was determined by ELISA at monthly intervals after vector inoculation. Rats inoculated with Q0ZHG had undetectable amounts of GDNF. QHGD-inoculated rats produced detectable but decreasing amounts of GDNF up to 3 months after inoculation, but no detectable GDNF after 4 months. QLGD-inoculated rats produced 1–2 pg/mg GDNF up to 6 months after inoculation.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

8. Fig. 7
Intracranial inoculation of QLGD protects against continuous intoxication with low-dose MPTP. Mice receiving MPTP daily 5 days of 7 show a progressive reduction in the number of pellets retrieved using the forelimb contralateral to vector inoculation. Animals inoculated with QLGD stabilize over 3 months of MPTP intoxication. Animals that did not receive MPTP treatment continued to retrieve 10–12 pellets over the course of 3 months (data not shown). Means ± SEM; the difference between the two curves is statistically significant (P < 0.001) by repeated-measures ANOVA.
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

9. Fig. 8
Mice inoculated with QLGD showed a significant preservation of TH-IR neurons in the SN ipsilateral to inoculation 6 months after vector inoculation. Representative photomicrographs demonstrate the loss of TH-IR cells in QHGD-inoculated animals compared to the contralateral, unlesioned side. The ratio of number of neurons (ipsilateral/contralateral) was determined from five to seven sections per animal with five or six animals per group. Scale bar, 200 μm. Means ± SEM, *P <
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

10. Fig. 9
Mice inoculated with QLGD showed a significant preservation of TH-IR and DAT-IR terminals in the striatum. Systemic administration of MPTP reduces TH IR and DAT IR bilaterally. The side of vector inoculation is indicated with an arrow. Representative immunostained brain sections are shown. The amount of immunoreactivity was quantitated using a PC-based image analysis system and reported as relative optical density (ROD). The ratio was determined from five to seven sections per animal with five or six animals per group. Means ± SEM, *P <
Molecular Therapy  , 67-75DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions