Figure 5

Qualitative and quantitative analysis of immunohistochemical staining for AR. Representative micrographs of control (A-C) and flutamide-treated boars (D-O) and a histogram of AR staining intensity expressed as relative optical density (ROD) of diaminobenzidine brown reaction products (P). Caput epididymis. In the control group (A) very strong intensity of AR staining is seen in the nuclei of principal cells (bold arrow), moderate to strong staining is seen in basal cells (arrow) and in stromal cells (arrowhead). In flutamide-treated males moderate to strong staining is seen in the nuclei of principal cells, weak staining is visible in basal cells and stromal cells (D, G, J, M). Note a slight decrease in the staining intensity in principal and basal cells, while in stromal cells a marked decrease is seen (F, I, L, O) Corpus epididymis. In the control group (B) strong intensity of AR staining in the nuclei of principal, basal, and stromal cells is seen. Note a marked decrease in the staining intensity in all cell types after flutamide-treatment (E, H, K, N) Cauda epididymis. In the control group (C) moderate to strong intensity of AR staining in the nuclei of principal and stromal cells and weak staining in basal cells is seen. Note a significant decrease in the staining intensity in principal and basal cells after flutamide exposure (F, I, L, O). In the control tissue section, no immunostaining for ARs is observed when the incubation was performed without the primary antibody (insert in A). principal epithelial cells - large arrows, basal epithelial cells - arrows, stromal cells - arrowheads. Nomarski interference contrast. Scale bar represents 20 μm. Values are means ± SD. Asterisks indicate significant differences between the respective control group and groups treated with flutamide prenatally (GD20, GD80) and postnatally (PD2, PD90) (n = 3 for each group). Statistical significance: *p < 0.05.