Table 3.
Confirmation by quantitative PCR of differential expression of noradrenergic neuron-relevant genes.
| Genes | Unigene Number | LongSAGE | Quantitative PCR (ratio KO/WT) | |||
| WT | KO | Day 7 NCC | Adult LC | Adult SCG | ||
| TH | Mm.1292 | 2 | 9 | 4.2 ± 0.4 (p = 0.001) | 2.4 ± 0.2 (p = 0.04) | 4.4 ± 0.2 (p = 0.03) |
| DBH | Mm.167781 | 4 | 14 | 4.1 ± 0.5 (p = 0.004) | 3.1 ± 0.2 (p = 0.000) | 6.8 ± 0.8 (p = 0.001) |
| MAO-A | Mm.21108 | 2 | 0 | 0.6 ± 0.0 (p = 0.006) | 0.5 ± 0.0 (p = 0.002) | 0.9 ± 0.1 (p = 0.04) |
| COMT | Mm.100940 | 0 | 1 | 1.1 ± 0.0 (p = 0.1) | 1.7 ± 0.5 (p = 0.2) | 1.4 ± 0.1 (p = 0.05) |
| Htr3a | Mm.4831 | 1 | 11 | 2.6 ± 0.6 (p = 0.004) | 3.5 ± 0.3 (p < 0.0001) | 4.0 ± 0.3 (p = 0.001) |
| CART | Mm.75498 | 12 | 22 | 2.0 ± 0.2 (p = 0.004) | 3.9 ± 0.2 (p < 0.0001) | 4.7 ± 0.3 (p = 0.001) |
| NET | Mm.57040 | 1 | 0 | Not detected in KO | Not detected in KO | Not detected in KO |
| Pja2 | Mm.41711 | 1 | 12 | 7.0 ± 1.8 (p = 0.04) | N/A | N/A |
| Numb1 | Mm.458153 | 0 | 7 | 3.5 ± 0.5 (p = 0.006) | N/A | N/A |
| Hdac2 | Mm.19806 | 12 | 1 | -6.97 ± 1.8 (p = 0.021) | N/A | N/A |
| Tlx2 | Mm.37 | 0 | 9 | 7.63 ± 1.2 (p = 0.04) | N/A | N/A |
For determining mRNA levels, the 2-ΔΔCT method was used as described. For a given target, ΔCT was computed by subtracting CT for HPRT from each primer pair. CT and ΔΔC were computed by subtracting ΔCT for WT from ΔCT for NETKO. The difference of the expression level for each gene was expressed as 2-ΔΔCT. Statistical analysis was performed using the Student's t test. qPCR data are presented as mean ± S.E.M., and significant differences between KO and WT reported at the p < 0.05 level; n = 3.