Relative mRNA expression of steroidogenic enzymes in the chick CAM.

<p>RT-qPCR analysis of mRNA coding for StAR, P450scc, P45017α, 3β-HSD and 17β-HSD on chick embryonic days 8 (n = 10), 10 (n = 17), 12 (n = 14), 14 (n = 10), 16 (n = 14), and 18 (n = 12). Data are reported as relative mRNA expression and represent mean normalized mRNA transcript number in copies/µL±SEM. 3β-HSD increased (F_{5, 68} = 89.282, p<0.0001) and 17β-HSD decreased (F_{5, 68} = 16.027, p<0.0001) significantly between embryonic day 8 and day 18.</p

Progesterone synthesis in the chick CAM.

<p>CAM sections were incubated in culture media for 2, 4, or 8 hours either with (circles) or without (squares) cholesterol (plus cAMP) as a precursor. Concentration of P4 in the culture media is represented as pg/ml of P4 per g of CAM tissue (pg/ml/g). Addition of precursor significantly increased the concentration of P4 in the culture media (F_{1, 58} = 46.917 p<0.0001) with a significant interaction between time of incubation and addition of precursor (F_{2, 58} = 3.709, p = 0.0305). To determine background and cross-reactivity of the P4 assay, controls consisting of only cholesterol and cAMP were incubated for 8 hours with an average P4 concentration of 0.337 pg/ml/g±0.423 SEM (not shown).</p

PR-ab and ERα mRNA expression and PR-ab immunolocalization.

<p>(A) RT-qPCR analysis of mRNA coding for PR-ab and ERα on chick embryonic days 8 (n = 10), 10 (n = 17), 12 (n = 14), 14 (n = 10), 16 (n = 14), and 18 (n = 12). Data are reported as relative mRNA expression and represent mean normalized mRNA transcript number in copies/µL±SEM. PR-ab (F_{5, 68} = 15.897, p<0.0001) and ERα (F_{5, 66} = 14.432, p<0.0001) increased significantly between embryonic day 8 and day 18. (B, C), PR-ab immunohistochemistry of embryonic day 18 CAM. (B) PR-ab positive section. Nuclear staining of PR-ab is localized predominately to the chorionic epithelium (c), and epithelial cells of blood vessels (bv). Positive nuclear staining is also present in allantoic epithelium (a), and mesenchyme (m). (C) Negative control of corresponding CAM section incubated without primary PR-ab antibody does not show specific nuclear staining. Scale bar represents 10 microns.</p