Crucial roles of the mesenchymal androgen receptor in Wolffian duct development
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Dec 18, 2023 version files 34.90 MB
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README.md
Abstract
Wolffian duct maintenance and differentiation is predominantly driven by the androgen action, which is mediated by the androgen receptor (AR). It is well established that the mesenchyme indicates the fate and differentiation of epithelial cells. However, in vivo developmental requirement of mesenchymal AR in Wolffian duct development is still undefined. By designing a mesenchyme-specific Ar knockout (ARcKO), we discovered that the loss of mesenchymal Ar led to the bilateral or unilateral degeneration of caudal Wolffian ducts and cystic formation at the cranial Wolffian ducts. Ex vivo culture of ARcKO Wolffian ducts invariably resulted in bilateral defects, suggesting that some factor(s) originating from surrounding tissues in vivo might promote Wolffian duct survival and growth even in the absence of mesenchymal Ar. Mechanistically, we found cell proliferation was significantly reduced in both epithelial and mesenchymal compartments; but cell apoptosis was not affected. Transcriptomic analysis by RNA-seq of E14.5 mesonephroi revealed 131 differentially expressed genes. Multiple downregulated genes (Top2a, Wnt9b, Lama2 and Lamc2) were associated with morphological and cellular changes in ARcKO male embryos (i.e. reduced cell proliferation and decreased number of epithelial cells). Mesenchymal differentiation into smooth muscle cells that are critical for morphogenesis was also impaired in ARcKO male embryos. Taken together, our results demonstrate the crucial roles of the mesenchymal AR in Wolffian duct maintenance and morphogenesis in mice.
README: Crucial roles of the mesenchymal androgen receptor in Wolffian duct development
https://doi.org/10.5061/dryad.5dv41nscp
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Supplementary figure 1. Coiling of the contralateral epididymis was decreased in ARcKO male embryos with the unilateral cystic epididymis. (A) Brightfield images of PND0 epididymides of the control and ARcKO groups. Regions outlined by dashed lines were enlarged for illustrating measurement of the number of turns. (B) Quantification of the average number of turns per 1 mm. Results were shown as mean ± SD and analyzed by two-tailed unpaired Student’s t-test. N=5 for control group, N=4 for ARcKO group.
Supplementary figure 2. Immunohistochemical staining of AR in PND0 epididymides from control, unaffected side of ARcKO, and affected side of ARcKO embryos.
Supplementary figure 3. Characterization of Osr2Cre+; Rosa26tdTomato+. (A) Whole mount immunofluorescent staining of tdTomato expression in Osr2Cre+; RosatdTomato+ male mesonephros and testis at E12.5. (B) Immunofluorescent staining of tdTomato expression in a representative section of Osr2Cre+; RosatdTomato+ male testis at PND0.
Supplementary figure 4. RT-PCR Quantification of Cyp17a1 and Hsd3b1 expression in control and ARcKO testes at E14.5. Results were shown as mean ± SD and analyzed by two-tailed Student’s t-test. N=4 for each group.