Norepinephrine transport-mediated gene expression in noradrenergic neurogenesis

BMC Genomics

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^-ΔΔC_T method was used as described. For a given target, ΔC_T was computed by subtracting C_T for HPRT from each primer pair. C_T and ΔΔC were computed by subtracting ΔC_T for WT from ΔC_T for NETKO. The difference of the expression level for each gene was expressed as 2^-ΔΔC_T. 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.

ISSN: 1471-2164