Abstract
BackgroundThe Sertoli cells act to induce testis differentiation and subsequent development in fetal and post-natal life which makes them key to an understanding of testis biology. As a major step towards characterisation of factors involved in Sertoli cell function we have identified Sertoli cell-specific transcripts in the mouse testis and have used the data to identify Sertoli cell-specific transcripts altered in mice lacking follicle-stimulating hormone receptors (FSHRKO) and/or androgen receptors (AR) in the Sertoli cells (SCARKO).ResultsAdult iDTR mice were injected with busulfan to ablate the germ cells and 50 days later they were treated with diphtheria toxin (DTX) to ablate the Sertoli cells. RNAseq carried out on testes from control, busulfan-treated and busulfan + DTX-treated mice identified 701 Sertoli-specific transcripts and 4302 germ cell-specific transcripts. This data was mapped against results from microarrays using testicular mRNA from 20 day-old FSHRKO, SCARKO and FSHRKO.SCARKO mice. Results show that of the 534 Sertoli cell-specific transcripts present on the gene chips, 85% were altered in the FSHRKO mice and 94% in the SCARKO mice (mostly reduced in both cases). In the FSHRKO.SCARKO mice additive or synergistic effects were seen for most transcripts. Age-dependent studies on a selected number of Sertoli cell-specific transcripts, showed that the marked effects in the FSHRKO at 20 days had largely disappeared by adulthood although synergistic effects of FSHR and AR knockout were seen.ConclusionsThese studies have identified the Sertoli cell-specific transcriptome in the mouse testis and have shown that most genes in the transcriptome are FSH- and androgen-dependent at puberty although the importance of FSH diminishes towards adulthood.
Highlights
The Sertoli cells act to induce testis differentiation and subsequent development in fetal and post-natal life which makes them key to an understanding of testis biology
Treatment of iDTR mice with busulfan caused a significant (P < 0.05, false discovery rate (FDR) = 0.05) change in the expression of 18,095 transcripts compared to control (6961 transcripts increased, 11,134 transcripts decreased; Additional file 4) while treatment with busulfan followed by diphtheria toxin (DTX) caused a significant change in levels of 5280 transcripts compared to busulfan alone (2891 transcripts increased and 2389 transcripts decreased; Additional file 4)
The only probable exceptions will be transcripts in another cell type which are highly dependent on the germ cells or Sertoli cells and which may be wrongly categorised as germ cell- or Sertoli cell-specific if their expression drops markedly after cell ablation
Summary
The Sertoli cells act to induce testis differentiation and subsequent development in fetal and post-natal life which makes them key to an understanding of testis biology. The development and function of the testis is critically dependent on the Sertoli cells These cells initially differentiate from the coelomic epithelium, at around embryonic day 11.5 (e11.5) in the mouse, and act to induce formation of the seminiferous tubules and the fetal Leydig cell population [1]. As previously [9], one confounding factor in identifying cell-specific transcript expression through cell ablation studies in the testis is the loss of germ cells coincident with Sertoli cell ablation [2]. To circumvent this problem, we have used busulfan to generate a germ cell-free mouse prior to Sertoli cell ablation [9]. This has the added advantage that comparison between normal mice and busulfan-treated animals allows identification of germ cell-specific transcripts
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