Supplementary material from "piRNA-associated proteins and retrotransposons are differentially expressed in murine testis and ovary of aryl hydrocarbon receptor deficient mice"

Previous studies suggested that the aryl hydrocarbon receptor (AhR) contributes to mice reproduction and fertility. However, the mechanisms involved remain mostly unknown. Retrotransposon silencing by piRNAs is essential for germ cell maturation and, remarkably, AhR has been identified as a regulator of murine B1-SINE retrotransposons. Here, using littermate AhR^+/+ and AhR^−/− mice, we report that AhR regulates the general course of spermatogenesis and oogenesis by a mechanism likely associated with piRNA-associated proteins, piRNAs and retrotransposons. piRNA-associated proteins MVH and Miwi are upregulated in leptotene to pachytene spermatocytes with a more precocious timing in AhR^−/− than in AhR^+/+ testes. piRNAs and transcripts from B1-SINE, LINE-1 and IAP retrotransposons increased at these meiotic stages in AhR-null testes. Moreover, B1-SINE transcripts colocalize with MVH and Miwi in leptonaema and pachynaema spermatocytes. Unexpectedly, AhR^−/− males have increased sperm counts, higher sperm functionality and enhanced fertility than AhR^+/+ mice. Contrary, piRNA-associated proteins and B1-SINE and IAP-derived transcripts are reduced in adult AhR^−/− ovaries. Accordingly, AhR-null female mice had lower numbers of follicles when compared with AhR^+/+ mice. Thus, AhR deficiency differentially affects testis and ovary development possibly by a process involving piRNA-associated proteins, piRNAs and transposable elements.