Figure 3

Qualitative and quantitative analysis of immunohistochemical staining for Cx43. Representative micrographs of control (A-C) and flutamide-treated boars (D-O) and a histogram of Cx43 staining intensity expressed as relative optical density (ROD) of diaminobenzidine brown reaction products (P). Caput epididymis. In the control group (A) and flutamide-treated groups (D, G, J, M) very weak signal for Cx43 or no Cx43 signal is seen between the basal and principal cells (arrow) and between the basal and stromal cells (arrowhead). Note a strong signal for Cx43 between neighboring principal cells in the initial segment of the epididymis (arrows) of GD20 and GD80 groups versus control group (inserts in A, D, G) Corpus epididymis. In the control group (B) very weak, punctate signal between the basal and principal cells (arrow) and moderate to strong signal between adjacent stromal cells are seen. In GD20, GD80 and PD2 a slight decrease in Cx43 signal between stromal cells or no decrease (PD90) compared to control is seen, respectively (E, H K, N) Cauda epididymis. In the control group (C) strong signal for Cx43 between stromal cells (arrowhead) and weak signal among basal cells (arrow) are seen. Note a significant decrease in the signal intensity in stromal cells of GD20 and PD2 groups (F, L), not in those of GD80 and PD90 (I, O). Negative control sections processed without primary antibody showed no staining (insert in B). principal epithelial cells - large arrows, basal epithelial cells - arrows, stromal cells - arrowheads. Nomarski interference contrast. 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