Behavioral and biochemical effects of antisense-mediated knockdown of
HuC gene expression in mouse brain. (a) Performance in
the probe session of the radial maze task in sham controls (TR) and in
mice daily infused during the training with fully sequence-degenerate
(TR + DN) or anti-HuC antisense (TR + AS) oligonucleotides
(n = 9 for TR animals and n = 8
for TR + DN and TR + AS animals). Data are expressed as means ±
SEM (**, P < 0.01, post hoc analysis
between DN-treated and AS-treated mice). (b) Effect of
the anti-HuC antisense treatment on HuC gene expression.
(Left) Real-time quantitative RT-PCR determination of
HuC mRNA steady-state levels normalized to GAPDH levels.
(Right) (Upper) Representative Western
blot of the overall ELAV-like immunoreactivity in the three groups of
mice compared with α-tubulin. (Lower) Means ±
SEM, n = 6 each group for both mRNA and protein
determinations. ****, P
< 0.001, post hoc analysis between DN-treated and AS-treated mice.
(c) Sequence specificity of the antisense-mediated HuC
gene expression down-regulation. Representative in situ
hybridizations showing hippocampal mRNA levels for the HuC and for the
phylogenetically related BRUNOL3 genes in the AS
ODN-treated and the two control mice groups. (d) Effect
of the anti-HuC antisense treatment on GAP-43 gene expression.
(Left) Real-time quantitative RT-PCR determination of
GAP-43 mRNA levels normalized to GAPDH levels in the hippocampi of the
three groups of mice (means ± SEM, n = 6 for
each group; **, P < 0.01, post hoc
comparison between DN-treated and AS-treated mice).
(Right) Densitometric analysis of GAP-43 mRNA levels in
the CA1 and CA3 hippocampal subfields from in situ
hybridization performed on brain slices of the same mice (means ±
SEM, data collected from five to seven mice from each group; *,
P < 0.05; ***,
P < 0.005).