Figure 5.

In vivo inhibition of KAP3 synthesis via icv administration of an AS ODN against KAP3 mRNA reduces basal GnRH release from the ME in vitro and delays the onset of puberty. A, Upper panel, Western blot showing that KAP3 AS ODN decreased KAP3 protein content in the MBH; middle panel, Western blot showing the lack of changes in β-tubulin (β-tub) content; this housekeeping protein was used as an internal control for normalization purposes; lower panel, quantitation of the changes shown in the upper panels. Each bar represents the mean of three Western blots, each using a pool of three MBH fragments. Vertical lines are sem. **, P < 0.01. AU, Arbitrary units. B, Effect of in vivo KAP3 AS ODN-mediated inhibition of KAP3 synthesis on the in vitro ability of the ME to release GnRH. Each point represents the mean ± sem of five to six independent observations. *, P < 0.05; **, P < 0.01 vs. AS ODN-treated group. C, Delay of female puberty caused by the disruption of KAP3 synthesis. The left panel depicts the percentage of animals in each group showing vaginal opening at different peripubertal ages; the right panel compares the mean age at vaginal opening in AS ODN- and SCR ODN-injected rats and sham-operated rats (n = 20 rats per group). Vertical lines are sem. **, P < 0.01 vs. the two control groups.