Table 2.
Tissue | Gene Symbol |
Postnatal Day 2 Cycles to Threshold |
Postnatal Day 8 Cycles to Threshold
|
||||||
---|---|---|---|---|---|---|---|---|---|
Baseline | 4 h | P | 2−ΔΔCt | Baseline | 4 h | P | 2−ΔΔCt | ||
Adrenal | Star | 23.71±0.23 | 23.65±0.10 | 0.800 | 0.81 | 23.98±0.15 | 22.84±0.14* | 0.001 | 2.10 |
Ldlr | 31.40±0.40 | 29.72±0.41* | 0.002 | 2.46 | 30.59±0.10 | 28.07±0.08* | 0.001 | 5.46 | |
Cyp11a | 23.20±0.10 | 23.44±0.08 | 0.097 | 0.76 | 23.55±0.08 | 23.88±0.27 | 0.282 | 0.90 | |
Cyp21a1 | 25.41±0.08 | 25.35±0.14 | 0.764 | 1.22 | 19.78±0.14 | 19.59±0.18 | 0.433 | 1.26 | |
Mc2r | 31.89±0.18 | 31.94±0.09 | 0.799 | ND | 32.28±0.39 | 32.46±0.49 | 0.780 | 1.34 | |
Hif1a | 26.83±0.13 | 27.11±0.09 | 0.110 | 0.78 | 26.27±0.04 | 26.77±0.21 | 0.061 | 0.67 | |
Npy | 19.62±0.16 | 19.10±0.20 | 0.087 | ND | 19.67±0.49 | 19.43±0.40 | 0.716 | ND | |
Anterior | Pomc | 25.38±0.07 | 25.61±0.19 | 0.159 | ND | 22.02±0.22 | 22.01±0.33 | 0.980 | ND |
Pituitary | Crhr1 | 34.29±0.26 | 35.00±0.13 | 0.255 | ND | 29.42±0.05 | 30.50±0.17* | 0.001 | ND |
Hsd11b1 | 32.65±0.19 | 32.80±0.24 | 0.822 | ND | 32.08±0.25 | 32.63±0.22 | 0.171 | ND | |
Hypothalamus | Crh | 34.37±0.19 | 34.26±0.24 | 0.731 | ND | 33.44±0.18 | 33.62±0.16 | 0.461 | ND |
Fos | 26.96±0.11 | 26.25±0.17* | 0.005 | ND | 26.76±0.07 | 26.57±0.17 | 0.317 | ND | |
Crhr1 | 27.43±0.07 | 27.73±0.10* | 0.032 | ND | 29.02±0.09 | 29.11±0.09 | 0.475 | ND | |
Nr3c1 (GR) | 23.73±0.06 | 23.89±0.09 | 0.190 | ND | 23.41±0.04 | 23.72±0.09* | 0.011 | ND | |
Nr3c2 (MR) | 27.33±0.06 | 26.94±0.22 | 0.115 | ND | 29.40±0.05 | 29.51±0.09 | 0.301 | ND | |
Hsd11b2 | 29.27±0.14 | 29.10±0.08 | 0.282 | ND | 29.44±0.17 | 29.53±0.08 | 0.640 | ND | |
Npy | 22.35±0.15 | 22.70±0.08 | 0.060 | ND | 22.70±0.10 | 22.76±0.11 | 0.686 | ND | |
Npy1r | 24.14±0.06 | 24.40±0.08* | 0.020 | ND | 25.95±0.08 | 26.13±0.07 | 0.138 | ND | |
Actb | 13.78±0.08 | 13.81±0.08 | 0.797 | ND | 16.57±0.08 | 16.73±0.11 | 0.282 | ND |
Whole adrenal glands (4–12 adrenals/sample) and anterior pituitary glands (2–6 pituitaries/sample) were pooled. Whole hypothalami were dissected and treated as one sample; n = 3–6 per tissue type for each gene of interest (at each age). RNA concentrations were normalized to 10–20 ng/μl, and each sample was assayed in triplicate for PCR analysis. A decrease in the cycle threshold (Ct) value at 4 h, as compared to baseline, translates into an increase in mRNA expression for that particular gene. Comparisons between age groups were not performed.
Significant difference in Ct value. Where indicated, the 2−ΔΔCt equation was used to calculate the fold change in target mRNA expression (vs. baseline). The 2−ΔΔCt equation uses a single reference gene (18S rRNA in this case) for the normalization of relative changes in target mRNA expression. See Table 1 for description of gene symbol abbreviations. ND, not determined.