TABLE 1.
qPCR analysis of nAChR subunit gene expression in rat adrenal medulla and pituitary gland
| Adrenal Medulla | Pituitary | Adrenal Medulla relative to Pituitary | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Gene | Ct | n | log102−(ΔCt) | Fold-less relative to α3 | Ct | n | log102−(ΔCt) | log102−(ΔΔCt) | Fold-difference |
| actin | 20.0 ± 0.7 | 6 | nd | nd | 20.1 ±0.09 | 6 | nd | nd | nd |
| GAPDH | 21.5 ± 1.2 | 6 | nd | nd | 21.0 ± 0.2 | 6 | nd | nd | nd |
| α2 | 37.1 ± 0.6 | 3 | −5.2 ± 0.1 | >30,000**** | 32.2 ± 1.7 | 3 | −3.5 ± 0.5 | −1.7 ± 0.4 | −44.7 ± 2.5**** |
| α3 | 22.1 ± 1.2 | 3 | −0.7 ± 0.4 | 1 | 34.8 ± 0.6 | 3 | −4.3 ± 0.2 | 3.7 ± 0.5 | 4,602 ± 3**** |
| α4 | 33.0 ± 0.8 | 3 | −3.9 ± 0.3 | >1,500**** | 29.6 ± 0.2 | 3 | −2.0 ± 0.04 | −1.9 ± 0.3 | −89.7 ± 2.0**** |
| α5 | 27.1 ± 1.0 | 3 | −2.2 ± 0.3 | 32 ± 1.6**** | 35.0 ± 1.4 | 3 | −4.4 ± 0.4 | 2.2 ± 0.6 | 163 ± 4**** |
| α6 | 39.6 ± 0.6 | 2 | −5.9 ± 0.2 | >150,000**** | 38.1 ± 1.6 | 3 | −5.3 ± 0.5 | −0.8 ± 0.4 | −4.2 ± 2.3 ns |
| α7 | 24.0 ± 0.7 | 3 | −1.2 ± 0.2 | 3.8 ± 1.6ns | 27.5 ± 0.7 | 3 | −2.2 ± 0.2 | 0.9 ± 0.3 | 7.7 ± 2.1* |
| α9 | 38.2 ± 0.5 | 2 | −5.2 ± 0.4 | >30,000**** | 30.0 ± 0.6 | 3 | −2.8 ± 0.2 | −2.4 ± 0.2 | −228 ± 1.5**** |
| α10 | 37.4 ± 0.5 | 3 | −5.0 ± 0.08 | >20,000**** | 31.0 ± 0.6 | 3 | −3.1 ± 0.2 | −1.9 ± 0.2 | −70.8 ± 1.5**** |
| β2 | 25.6 ± 0.8 | 3 | −1.4 ± 0.2 | 5.7 ± 1.6* | 24.5 ± 0.3 | 3 | −1.2 ± 0.04 | −0.3 ± 0.2 | −1.8 ± 1.5 ns |
| β3 | 39.3 ± 0.5 | 2 | −5.6 ± 0.08 | >90,000**** | 35.6 ± 0.9 | 3 | −4.5 ± 0.3 | −1.3 ± 0.06 | −13.5 ± 0.3** |
| β4 | 24.9 ± 0.9 | 3 | −1.2 ± 0.3 | 3.5 ± 1.6ns | 30.6 ± 0.4 | 3 | −3.0 ± 0.1 | 1.8 ± 0.3 | 61.2 ± 2.1**** |
Tissues from three animals were analyzed individually. Positive values for comparisons of gene expression between adrenal medulla and pituitary indicate greater relative abundance in the adrenal medulla. Significance was determined using a one-way ANOVA with a Holm-Šídák post hoc comparison test
not significant or p > 0.05
p ≤ 0.05
p ≤ 0.01
p ≤ 0.001
p ≤ 0.0001
‘±’ values indicate the SD; Ct, cycle threshold; fold-less and fold-difference values were converted from logarithmic units; ‘n’ values indicate number of tissue samples where the reaction reached threshold; inclusion of actin and GAPDH reactions in two separate runs resulted in higher n values and were included in each run to ensure plate-to-plate reproducibility of results.