Figure 9

VEGF antibody inhibits the SON proliferative response. Rats were implanted unilaterally with a cannula positioned over the SON and connected to an Alzet osmotic pump containing a blocking VEGF antibody (schematic drawing), osmotically stimulated during 6 days, and injected with BrdU 5 hours before their fixation. A-D: Stack confocal images (10 μm-thick) of sections double immunostained for BrdU and goat-IgG. In the SON homolateral to the cannula, the number of BrdU-labeled nuclei is clearly lower that in the SON contralateral to the cannula (A vs C). Immunostaining for goat-IgG indicates that the goat-IgG anti-VEGF antibody has diffused from the cannula to the homolateral SON (B), but not to the contralateral SON (D). Note that numerous BrdU-labeled nuclei are observed along the border of the lesional cavity formed by the cannula diffusing the anti-VEGF antibody (arrow in A). E: Quantitative evaluation of the number of BrdU-labeled nuclei detected within the SON of 6 days osmotically stimulated rats implanted with a cannula diffusing either the anti-VEGF antibody or the isotype IgG. Data indicate that cell proliferation is significantly inhibited in the SON homolateral to implanted with cannulas diffusing blocking anti-VEGF antibody, but not with cannulas diffusing goat-IgG. Data represent means ± SEM of counts made on at least 4 SON areas per rats in 4 rats per experiment. Double asterisk: P < 0.01, Mann-Whitney test, statistically different from the SON contra-lateral to the cannula. BrdU: bromodeoxyuridine; cl: contra-lateral to the cannula; hl: homo-lateral to the cannula; IgG: type G immunoglobulin; OC: optic chiasma; SON: supraoptic nucleus; VEGF: vascular endothelial growth factor. Scale bar: A-D = 100 μm.