Connexin 43 gene expression in male and female gonads of porcine offspring following in utero exposure to an anti-androgen, flutamide

The aim of this study was to show the effect of maternal exposure to flutamide on connexin 43 (Cx43) gene expression in testes and ovaries of 2-day-old piglets. Additionally, anogenital distance (AGD) was measured both in male and female offspring. Immunohistochemistry, Western blotting, and RT-PCR were performed to assess the immunoreactivity and the presence of Cx43 protein and itʹs mRNA, respectively. Following flutamide exposure strong immunostaining for Cx43 was observed between testicular Leydig cells, between granulosa cells of primary follicles, and between interstitial cells surrounding clusters of oocyte nests in the ovarian cortex as in the respective controls. Differences between the flutamide-treated groups and the controls obtained by qualitative immunohistochemistry were confirmed by quantitative image analysis (*P<0.05; **P<0.01). In Western blotting, Cx43 appeared as a band of 43 kDa, whereas electrophoresis revealed PCR products of the predicted sizes. Screening for Cx43 expression revealed the presence of a transcript, both in control and in flutamide-treated pigs. The AGD values differed significantly from the control (*P<0.05). Overall, since no obvious changes in gonad morphology were observed and the Cx43 signal was present in all the examined tissues, it seems likely that androgens acting through ARs are not involved in the control of Cx43 gene expression in neonatal pig gonads.