Organotypic cultures of prepubertal mouse testes: a method to study androgen action in sertoli cells while preserving their natural environment

Karel De Gendt, Chris McKinnell, Ariane Willems, Philippa T K Saunders, Richard M Sharpe, Nina Atanassova, Johannes V Swinnen, Guido Verhoeven

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Cluster analysis at Postnatal Day 8-20 of putative androgen-regulated genes in mice with Sertoli cell-selective knockout of the androgen receptor (SCARKO) has pinpointed three genes (Spinlw1, Gpd1, Drd4) with an expression pattern strongly resembling that of Rhox5, the definitive Sertoli cell (SC) androgen-regulated gene. We used organotypic testis cultures from Day 8 mice to study control of these genes by (anti)androgens and follicle-stimulating hormone (FSH). Testis morphology and androgen induction of the studied genes were preserved for 48 h. Preincubation with ketoconazole for 24 h to block endogenous androgen production, followed by 24-h incubation with the synthetic androgen R1881, resulted in 45-, 5-, 19-, and 6-fold induction of mRNA levels of Rhox5, Spinlw1, Gpd1, and Drd4, respectively. However, noticeable differences in control of the studied genes were observed. Rhox5 and Spinlw1 were fully induced by R1881 in the continuous (48 h) presence of ketoconazole, whereas only marginal effects were observed on expression of Gpd1 and Drd4. Similarly, FSH only marginally affected expression of Rhox5 and Spinlw1, whereas it markedly increased Gpd1 and Drd4 expression. Explant cultures of SCARKO testes confirmed the differential effects of FSH on the studied genes and, for Gpd1, showed that the effect did not depend on a functional androgen receptor in SC, whereas this was essential for the effects of FSH on Drd4. In conclusion, organotypic cultures represent the first in vitro approach to preserving androgen responsiveness of putative SC-expressed genes. This approach facilitates detailed analysis of their regulation in ways not possible in vivo.
Original languageEnglish
Pages (from-to)1083-1092
Number of pages10
JournalBiology of Reproduction
Volume81
Issue number6
Early online date8 Jul 2009
DOIs
Publication statusPublished - 1 Dec 2009

Keywords

  • analysis of variance
  • androgens
  • animals
  • apoptosis
  • culture media, conditioned
  • gene expression profiling
  • glycerolphosphate dehydrogenase
  • homeodomain proteins
  • immunohistochemistry
  • male
  • mice
  • organ culture techniques
  • organ size
  • proteins
  • RNA, messenger
  • receptors, androgen
  • receptors, dopamine D4
  • reverse transcriptase polymerase chain reaction
  • sertoli cells
  • testis
  • testosterone
  • transcription Factors

Fingerprint Dive into the research topics of 'Organotypic cultures of prepubertal mouse testes: a method to study androgen action in sertoli cells while preserving their natural environment'. Together they form a unique fingerprint.

  • Cite this

    De Gendt, K., McKinnell, C., Willems, A., Saunders, P. T. K., Sharpe, R. M., Atanassova, N., Swinnen, J. V., & Verhoeven, G. (2009). Organotypic cultures of prepubertal mouse testes: a method to study androgen action in sertoli cells while preserving their natural environment. Biology of Reproduction, 81(6), 1083-1092. https://doi.org/10.1095/biolreprod.109.078360