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Transduction characteristics of adeno-associated virus vectors expressing cap serotypes 7, 8, 9, and Rh10 in the mouse brain  Cassia N. Cearley, John.

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Presentation on theme: "Transduction characteristics of adeno-associated virus vectors expressing cap serotypes 7, 8, 9, and Rh10 in the mouse brain  Cassia N. Cearley, John."— Presentation transcript:

1 Transduction characteristics of adeno-associated virus vectors expressing cap serotypes 7, 8, 9, and Rh10 in the mouse brain  Cassia N. Cearley, John H. Wolfe  Molecular Therapy  Volume 13, Issue 3, Pages (March 2006) DOI: /j.ymthe Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

2 FIG. 1 Comparison of vector and enzyme distribution resulting from AAV 7, 8, 9, or Rh10 HβH injections at 2 months postinjection. Animals (n = 3 or 4/serotype) were injected in the hippocampus, thalamus, cortex, and striatum, 1 μl per injection site. Shown are (B, D, F, H) the enzyme histochemistry and (A, C, E, G) corresponding in situ hybridization (ISH) of different areas of the brain. The level of the slice is given as its rostral–caudal coordinate relative to bregma [32], and the levels of injection are represented by asterisks. Injections of the different serotypes resulted in dramatically different transduction patterns and enzyme distribution. While injections of AAV Rh10 resulted in the most enzyme distribution (H), injections of AAV 9 showed the most vector distribution to the contralateral side (E) with ISH-positive cells in the contralateral hippocampus (E, arrow). Injections of AAV 7 or 8 also resulted in high levels of vector-and enzyme-positive cells. A high-magnification inset showing AAV 8 transduction of hippocampal cells is shown. Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

3 FIG. 2 Cell-type specificity of the individual serotypes. Fluorescence in situ hybridization for hGUSB mRNA was performed on brain sections from mice injected with AAV 7, 8, 9, or Rh10, followed by incubation in antibodies to PLP, GFAP, or Map2ab. Cy3 (red) was used for GUSB detection and FITC (green) was used for PLP, GFAP, and Map2ab detection. Pictures were taken by confocal microscope. All cells positive for vector mRNA were colocalized with Map2ab-positive cells, but not PLP-or GFAP-positive cells, indicating that the vectors transduced neurons, but not oligodendrocytes or astrocytes. Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

4 FIG. 3 The AAV 9 vector genome undergoes transport following a unilateral injection into the hippocampus. Animals (n = 3) were injected in the hippocampus and then analyzed 2 months postinjection for vector and enzyme distribution. Shown are the enzyme histochemistry and corresponding in situ hybridization. A unilateral injection resulted in both enzyme-and vector-positive cells in the contralateral hippocampus, which appear to have been transported via the dorsal hippocampal commissure. Vector-and enzyme-positive cells were also found in the septal nuclei and ipsilateral entorhinal cortex, both projection sites of the hippocampus. Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions

5 FIG. 4 Reversal of storage pathology in the MPS VII mouse following injections of AAV 9 HβH into the CNS. Mice (n = 3) were injected in the cortex, striatum, hippocampus, and thalamus at a volume of 1 μl per injection site and analyzed 2 months postinjection. Brains were embedded in JB4 resin, sectioned at a thickness of 1 μm, and stained using toluidine blue, which highlights storage vacuoles. Reversal of storage lesions was seen throughout the brain, compared to uninjected MPS VII control mice. Representative sections are shown with examples of storage vacuoles demarcated by arrows. Molecular Therapy  , DOI: ( /j.ymthe ) Copyright © 2005 The American Society of Gene Therapy Terms and Conditions


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