DOI: 10.5061/dryad.5dv41nscp
title: Crucial roles of the mesenchymal androgen receptor in Wolffian duct development

description: 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.
description: # Crucial roles of the mesenchymal androgen receptor in Wolffian duct
 development Give a brief
 summary of dataset contents, contextualized in experimental procedures and
 results. ## Description of the data and file structure **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.