FIG. 5.

Representative photomicrographs of sections of various mouse tissues 6 weeks after tail vein injection of AAV8-EF1α-nlslacZ at a dose of 7.2 × 10¹² vg/mouse (A to D) or 3 weeks after tail vein injection of AAV8-CMV-lacZ at a dose of 3.0 ×10¹¹ or 7.2 × 10¹² vg/mouse (E). The sections were either X-Gal stained (A, B, D, and E) or stained with designated antibodies (C). (A) Tissue distribution of β-galactosidase-positive cells: lu, lung; h, heart; s, spleen; k, kidney; i, intestine; t, testis; p, pancreas; and m, skeletal muscle (quadriceps). The top row represents tissues from a mouse injected with excipient only, while the bottom row shows samples from vector-injected mice. (B) Brain transduction with rAAV8. (a) Cerebral cortex. Positive cells are scattered throughout the region. (b) Hippocampus. Positive cells are observed in both granule and pyramidal cell layers. (c) Striatum. (d) Amygdala. (e) Hypothalamus. β-Galactosidase-positive neurons and glial cells are clustered in the arcuate nucleus and median eminence. Some ependymal cells of the third ventricle are also positive. (f) Cerebellum. Purkinje cells are regionally well transduced. (C) Confocal microscopy to assess colocalization of β-galactosidase and either NeuN (a marker for neurons) or GFAP (a marker for astrocytes) to determine rAAV8-transduced cell types in the cerebral cortex of the brain. Both neurons and glial cells were transduced with rAAV8. Scale bars, 5 μm. (D) Transduction of vascular smooth muscle cells in the walls of a branch of the coronary artery (a) and a branch of the splenic artery (b). (E) Tissue distribution of β-galactosidase-positive cells in mice injected with AAV8-CMV-lacZ via the tail vein. The vector doses (vg/mouse) are indicated above the pictures. The section of the pancreas was also stained with anti-insulin antibody (brown cells). Scale bars (duplicated lines), 250 μm. lv, liver; t.a., tibialis anterior limb muscle. The tissues in panels A, B, D, and E were counterstained with nuclear fast red or light hematoxylin. Scale bars represent 100 μm unless otherwise noted.