Figure 5. Analysis of Thy1 expression in optic nerve crush-resistant ganglion cells of Bax^-/- mice

Analysis of Thy1 expression in retinal ganglion cells in Bax^+/+ and Bax^-/- mice after optic nerve crush. The change in Thy1 expression 1 week after optic nerve crush was monitored by in situ hybridization (A-D) and RPA analysis of equal amounts of total RNA (E). High magnification Nomarski images of the central retinal ganglion cell layer of a Bax^+/+ mouse control eye (A) and experimental eye 1 week after crush (B). Like the CB6F1 mice, optic nerve crush causes a decrease in the number of Thy1 positive cells in the retina before there is noticeable cell loss. C: A similar view of the central retina from a control Bax^-/- mouse, showing a typical increase in the number of ganglion cells and robust Thy1 staining. D: One week after optic nerve crush, Bax^-/- mice also show a decrease in Thy1 staining. Size bar, 10 mm. E: The change in Thy1 expression observed by in situ hybridization was also monitored by RPA of total RNA. Bax^-/- mice contain more Thy1 mRNA per mg of total RNA than wild-type littermates, but both groups of mice exhibit approximately 50% loss of the Thy1 mRNA 1 week after crush. No fragment was protected by the murine Thy1 probe in an equal mass of total RNA isolated from human retinoblastoma cells (hRB RNA).