Download presentation
Presentation is loading. Please wait.
Published byFerenc Vörös Modified over 5 years ago
1
The Lymphocytic Choriomeningitis Virus Envelope Glycoprotein Targets Lentiviral Gene Transfer Vector to Neural Progenitors in the Murine Brain Colleen S. Stein, Inês Martins, Beverly L. Davidson Molecular Therapy Volume 11, Issue 3, Pages (March 2005) DOI: /j.ymthe Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
2
Fig. 1 Transgene-expressing cells appear in the OB after injection of LCMV-pseudotyped lentiviral vector into the striatum. Three weeks after injection of LCMV/FIV-CMVntβ-gal (1.5 × 106 TU), numerous β-gal-positive nuclei (X-gal-stained blue) were detected (A) not only around the site of injection in the striatum (B, lower boxed area in A) and along the corpus callosum (C, upper boxed area in A), but also (D) in the SVZ/proximal RMS and (E) in the OB (F, boxed area in E). Images were captured at 100× (A), 200× (E), or 600× (B, C, D, F) magnification. The arrow in A indicates the direction of the needle path, which continues ventrally, and the line approximates the interface between the striatum and the corpus callosum. Arrowheads in B and C indicate examples of transgene-positive cells in the striatum. Arrows in C and D indicate examples of transgene-positive cells in the corpus callosum and SVZ/proximal RMS, respectively. v, ventricle; str, striatum; cc, corpus callosum. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
3
Fig. 2 Expression of transgene occurs along the neurogenic pathway from the SVZ to the OB. Three weeks after injection of LCMV/FIV-CMVβ-gal (3 × 106 TU) into the striatum, β-gal-expressing cells were apparent in the striatum and in the SVZ, along the RMS, and in the OB. This pattern was detected for mice injected at (A) 3 weeks of age and (B) 16 weeks of age. Higher power images show morphology suggestive of (C) astrocytes in the striatum and (D) neurons in the OB. Images were captured from 40-μm stained sections at 40× (A, B), 400× (C), or 300× (D) magnification. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
4
Fig. 3 Determination of neural cell types by confocal analysis of sections from LCMV/FIV-CMVβ-gal-injected mice. Three weeks or 7.5 weeks after LCMV/FIV-CMVβ-gal injection into the striatum, brain sections were dual stained for β-gal (green) and GFAP, class III β-tubulin, or NeuN (red) and analyzed by confocal microscopy. Merged images are shown, and overlap of green and red fluorescence appears yellow. Colocalization of β-gal and GFAP is prominent in the (A) striatum and (B) SVZ, but is minimal in the (C) proximal RMS (arrowheads) and is absent from the (D) horizontal, or more distal, RMS. In the RMS, β-gal expression colocalizes with class III β-tubulin (E and F, 3 weeks postinjection; G, 7.5 weeks postinjection). (H) In the OB, expression of β-gal overlaps with NeuN. Scale bars, 20 μm. Images shown are 1.7- to 2.0-μm (B, C, D, E, H) or to 0.44-μm (A, F, G) slices from captured z series. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
5
Fig. 4 Proliferation occurs after LCMV-directed transduction. LCMV/FIV-CMVβ-gal (3 × 106 TU) was injected into the striatum on day 0, BrdU was injected daily ip from day 1 to day 18, and mice were sacrificed on day 19. (A and B) Confocal analysis of the OB shows β-gal-positive cells (green, arrowheads) that also stain positive for NeuN (blue) and BrdU (red), as seen in the merged images (right). (C) Similarly in the RMS, a β-gal-positive cell (green, arrowhead) is also positive for class III β-tubulin (blue) and BrdU (red). Scale bars, 20 μm. Images shown are (A) or 0.38-μm (B, C) slices from captured z series. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
6
Fig. 5 LCMV-pseudotyped FIV transduces GFAP-expressing neuronal precursor/stem cells. Three weeks after injection of LCMV/FIV-GFAPcre into the striatum of ROSA26 Cre reporter mice, β-gal-expressing cells were detected in the (A) RMS and (B) OB. In the RMS, β-gal-positive cells (green, arrowheads) are also class III β-tubulin positive (red), as seen in the merged image (right). In the OB, β-gal-positive cells (green, arrowheads) are also NeuN positive (red), as seen in the merged image (right). Scale bars, 20 μm. Images shown are 1.0- (A) and 0.44-μm (B) slices from captured z series. Molecular Therapy , DOI: ( /j.ymthe ) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions
Similar presentations
© 2024 SlidePlayer.com. Inc.
All rights reserved.