Figure 2

Gestational exposure to CD decreases the number of undifferentiated SG in the F3 generation males and leads to a decrease in spermatozoa number in F1 and F3. (A) Representative images of testes sections from the control (left panel) and CD lineage (right panel) animals: Sertoli and SG cells were immunostained using anti-GATA1 (red) or anti-ZBTB16 (green) antibodies, respectively. The ZBTB16 antibody staining of Leydig cells located outside of seminiferous tubules is non-specific, scale bar 150 µm. A quantitative analysis of (B) Sertoli cells and (C) SG was performed by manually counting the cells at stage VII of the seminiferous epithelium in F3 mouse testes. The contour of each tubule section was measured using ImageJ. The values shown indicate the cell counts per micrometre of tubule circumference. (D) The ratio of the number of Sertoli cells per SG is also indicated (**p < 0.01, n = 7 control, n = 6 CD, t-test). The immunostaining of the testis sections was performed as described in the Methods section. (E) Spermatozoa numbers in F1 and F3 have decreased. Spermatozoa were counted as described in the Methods section, by using epididymis of at least 8 biological replicates for control and CD and data presented as normalised value compared to control +/−SEM, (F1: p = 3.69E-4, n = 9 control and CD; F3: p = 1.64E-5, n = 8 control, n = 9 CD, t-test).