Fetal Leydig Cell Development Research Articles

Nuclear receptors Sf1 and Dax1 function cooperatively to mediate somatic cell differentiation during testis development

Mutations of orphan nuclear receptors SF1 and DAX1 each cause adrenal insufficiency and gonadal dysgenesis in humans, although the pathological features are distinct. Because Dax1 antagonizes Sf1-mediated transcription in vitro, we hypothesized that Dax1 deficiency would compensate for allelic loss of Sf1. In studies of the developing testis, expression of the fetal Leydig cell markers Cyp17 and Cyp11a1 was reduced in heterozygous Sf1-deficient mice at E13.5, consistent with dose-dependent effects of Sf1. In Sf1/Dax1 (Sf1 heterozygous and Dax1-deleted) double mutant gonads, the expression of these genes was unexpectedly reduced further, indicating that loss of Dax1 did not compensate for reduced Sf1 activity. The Sertoli cell product Dhh was reduced in Sf1 heterozygotes at E11.5, and it was undetectable in Sf1/Dax1 double mutants, indicating that Sf1 and Dax1 function cooperatively to induce Dhh expression. Similarly, Amh expression was reduced in both Sf1 and Dax1 single mutants at E11.5, and it was not rescued by the Sf1/Dax1 double mutant. By contrast, Sox9 was expressed in single and in double mutants, suggesting that various Sertoli cell genes are differentially sensitive to Sf1 and Dax1 function. Reduced expression of Dhh and Amh was transient, and was largely restored by E12.5. Similarly, there was recovery of fetal Leydig cell markers by E14.5, indicating that loss of Sf1/Dax1 delays but does not preclude fetal Leydig cell development. Thus, although Sf1 and Dax1 function as transcriptional antagonists for many target genes in vitro, they act independently or cooperatively in vivo during male gonadal development.

Pdgfr-alpha mediates testis cord organization and fetal Leydig cell development in the XY gonad.

During testis development, the rapid morphological changes initiated by Sry require the coordinate integration of many signaling pathways. Based on the established role of the platelet-derived growth factor (PDGF) family of ligands and receptors in migration, proliferation, and differentiation of cells in various organ systems, we have investigated the role of PDGF in testis organogenesis. Analysis of expression patterns and characterization of the gonad phenotype in Pdgfr-alpha(-/-) embryos identified PDGFR-alpha as a critical mediator of signaling in the early testis at multiple steps of testis development. Pdgfr-alpha(-/-) XY gonads displayed disruptions in the organization of the vasculature and in the partitioning of interstitial and testis cord compartments. Closer examination revealed severe reductions in characteristic XY proliferation, mesonephric cell migration, and fetal Leydig cell differentiation. This work identifies PDGF signaling through the alpha receptor as an important event downstream of Sry in testis organogenesis and Leydig cell differentiation.

Transplantation of Newborn Rat Testis Under the Kidney Capsule of Adult Host as a Model to Study the Structure and Function of Leydig Cells

Newborn rat testis was transplanted under the kidney capsule of adult castrated and uncastrated male rats to develop and characterize a model system for studies on Leydig cell development. Two weeks after transplantation, the number of Leydig cells and the size of their nuclei in the transplants had increased. Secretion of testosterone was indicated by increased seminal vesicle weights and decreased pituitary LH in the castrated host animals. Pituitary FSH content increased significantly in the uncastrated animals with transplants, which suggested production of an FSH-stimulating factor. Cells with the morphologic features characteristic of fetal- and adult-type Leydig cells were observed in the transplants. The seminiferous tubules with spermatocytes, incipient lumina, and significantly larger average diameters showed more advanced development than those in the normal 2-week-old testis. By the present morphologic and functional criteria, the kidney subcapsular transplantation technique provides a suitable model for studies of fetal and adult Leydig cell development.

Leydig cell differentiation in the prepubertal rabbit testis.

The pattern of Leydig cell differentiation in the postnatal rabbit was studied. The adult generation of Leydig cells appears at 5–6 weeks of age, with a sharp rise in the percentage of Leydig cells in the interstitial tissue at 6–7 weeks. There is a further steady, although less steep increase up to 9–10 weeks and a second sharp rise at 10–11 weeks to reach adult levels. Mitosis is active in the interstitial tissue between 5 and 7 weeks, then falls off sharply and is absent after 9 weeks. The increase in relative numbers of Leydig cells is therefore only partly a result of cell multiplication and primarily due to differentiation of cells in the testicular interstitium. Partially differentiated cells comprise a significant and relatively constant proportion of the interstitial cell population throughout the postnatal period, as occurs during fetal Leydig cell development, indicating similarity in the pattern of differentiation of the two Leydig cell generations. However, the differentiation process occurs over several weeks postnatally, as opposed to only a few days in the fetus.