1. Volume 11, Issue 1, Pages 89-95 (January 2005)
Lentiviral-Mediated Delivery of Bcl-2 or GDNF Protects against Excitotoxicity in the Rat Hippocampus 
Liang-Fong Wong, G. Scott Ralph, Lucy E. Walmsley, Alison S. Bienemann, Stephen Parham, Susan M. Kingsman, James B. Uney, Nicholas D. Mazarakis 
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
Transduction of cultured hippocampal cells by EIAV-Bcl-2 and EIAV-GDNF and their neuroprotective effects. E18 hippocampal cultures were transduced with (A) EIAV-LacZ, (B) EIAV-Bcl-2, or (C) EIAV-GDNF at an m.o.i. of 10 and the expression of the appropriate transgenes was determined using antibodies against β-galactosidase, Bcl-2, or GDNF, respectively (all indicated in green in A–C). Colocalization with a neuronal marker was determined using antibody against MAP-2 (red), while all cells were stained with DAPI (blue). Approximately 50% of total cells were transduced. (D) The percentage of cells transduced by EIAV-LacZ, EIAV-Bcl-2, or EIAV-GDNF that colocalized with MAP2 or a glial marker, GFAP, was assessed by counting the number of transduced cells that express MAP2 (MAP2+) or GFAP (GFAP+) and expressing it as a percentage of the total transduced cells. (E) Protection against glutamate excitotoxicity by EIAV-Bcl-2 and EIAV-GDNF in hippocampal cultures. Control hippocampal cultures transduced by EIAV-LacZ underwent cell death after exposure to 100 μM or 1 mM glutamate. In contrast, EIAV-Bcl-2- and EIAV-GDNF-transduced hippocampal cells were protected against glutamate toxicity. Scale bars (A–C) represent 50 μm.
Molecular Therapy  , 89-95DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

3. Fig. 2
Transduction of rat hippocampus by lentiviral vectors. (A) EIAV-LacZ pseudotyped with rabies-G was injected into the rat hippocampus and expression of the marker gene was assessed by X-gal staining. EIAV-LacZ pseudotyped with rabies-G demonstrated good expression in the CA1 pyramidal neurons. Similarly, good transduction was observed with (B) EIAV-Bcl-2 and (C) EIAV-GDNF. (D) In the EIAV-LacZ animals, the majority of β-galactosidase expression (green) occurred in neurons, which express NeuN (red). Scale bars represent 100 μm (A) and 50 μm (B–D).
Molecular Therapy  , 89-95DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

4. Fig. 3
Neuroprotection by EIAV-Bcl-2 against NMDA excitotoxicity in the rat hippocampus. (A) Injection of NMDA induced an excitotoxic lesion (indicated by arrowhead) in the CA1 layer of the hippocampus in control EIAV-LacZ-transduced rats. Lesion results in the loss of NeuN staining and widespread cellular damage to the CA1 layer of the hippocampus. (B) In contrast an insignificant lesion was induced in EIAV-Bcl-2-transduced rats. (C) Using Fluorojade-B staining a large number of fluorescent neurons in the CA1 were observed in the NMDA-induced lesion in EIAV-LacZ animals, whereas (D) only a few fluorescent cells could be detected in EIAV-Bcl-2 animals. (E) Quantitative assessment of excitotoxic lesion in both groups indicated that EIAV-Bcl-2 significantly protected hippocampal neurons from NMDA-induced toxicity. *P < 0.05 (Student's unpaired t test). Scale bar represents 1 mm (A and B) and 50 μm (C and D).
Molecular Therapy  , 89-95DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

5. Fig. 4
EIAV-GDNF protects rat hippocampal neurons against NMDA excitotoxicity. (A) An excitotoxic lesion (indicated by arrowhead) in the CA1 layer of the hippocampus was produced by NMDA injection in control EIAV-LacZ-transduced rats. In this lesion, there was significantly reduced NeuN staining compared to the contralateral control PBS-injected side. (B) In EIAV-GDNF-transduced rats, only a small area of lesion was induced by NMDA as indicated by the presence of NeuN staining. (C) A large number of CA1 neurons in the lesion were stained with Fluorojade-B in EIAV-LacZ animals, whereas (D) only a few fluorescent cells could be detected in EIAV-GDNF animals. (E) Quantitative assessment of excitotoxic lesion in both groups showed that EIAV-GDNF significantly reduced the lesion size compared to EIAV-LacZ rats. *P < 0.05 (Student's unpaired t test). Scale bar represents 1 mm (A and B) and 50 μm (C and D).
Molecular Therapy  , 89-95DOI: ( /j.ymthe )
Copyright © 2004 The American Society of Gene Therapy Terms and Conditions