Histogenesis of the Gubernaculum During Testis Descent in the Goat Fetus (Capra marghoz) / Histogénesis del Gubernaculum Durante el Descenso Testicular en el Feto de Cabra (Capra marghoz)

SUMMARY: Testicular descent is a complex process that only occurs in mammals. The role of the gubernaculum during testicular descent has been explained mainly by its capacity for dilatation and contraction. This study tried to investigate the changes in the structure of the fibers and cells of the gubernaculum in different age levels of testicular descent in goat fetuses. Embryo samples were collected and grouped in such a way that 60 male goat fetuses were obtained from 100 pregnant does (Capra marghoz). The samples were classified based on the average length (CRL) of the used embryos into 6 age groups. Tissues of the gubernaculum were stained using Masson's Trichrome method to observe collagen fibers under light microscopy. In the present study, growth and orientation of collagen fibers of gubernaculum were observed from the age of 51 days in a manner that the arrangement and order of fibroblasts and collagens to be associated with the onset of testicular migration order and collagen fibers until the end of the third month. Further, changes in the cell arrays and strings were observed after the age of 111 days in such a way that near the birth date, the gubernaculum converted into atrophy tissue. It can be said that from the beginning of the period of testicular descent until its completion, the tissue of the gubernaculum undergoes cellular changes, such as deformation and increase and secretion in connective fibers.

El descenso testicular es un proceso complejo que solo ocurre en los mamíferos. El papel del gubernaculum durante este proceso se ha explicado principalmente por su capacidad de dilatarse y contraerse. En este trabajo, se investigaron los cambios en la estructura de las fibras y células del gubernaculum en diferentes etapas del descenso testicular y edades en fetos de cabra. Se recolectaron muestras de embriones, agrupándose de manera que se obtuvieron 60 fetos de macho cabrío a partir de 100 hembras preñadas (Capra marghoz). Las muestras se clasificaron según la longitud media (CRL) de los embriones utilizados, dividiéndose en seis grupos de edad. Los tejidos del gubernaculum se tiñeron utilizando la técnica de Tricrómico de Masson para observar las fibras de colágeno bajo microscopía óptica. En el presente estudio, se observó el crecimiento y la orientación de las fibras colágenas del gubernaculum a partir de los 51 días de edad. La disposición y el orden de los fibroblastos y colágeno se asociaron con el inicio de la migración testicular, observándose las fibras colágenas hasta el final del tercer mes. Además, se detectaron cambios en las matrices y cadenas de células después de los 111 días de edad. Cerca de la fecha de nacimiento, el gubernaculum se convirtió en tejido atrofiado. En conclusión, desde el inicio hasta la finalización del período de descenso testicular, el tejido del gubernaculum sufre cambios celulares, como deformación y aumento de secreción en las fibras conectivas.

Expression and Role of INSL3 in the Fetal Testis.

Insulin-like peptide 3 (INSL3) is a small peptide hormone of the insulin-relaxin family which is produced and secreted by the fetal Leydig cells in the testes only. It appears to be undetectable in female fetuses. In the human fetus INSL3 synthesis begins immediately following gonadal sex determination at weeks 7 to 8 post coitum and the peptide can be detected in amniotic fluid 1 to 2 weeks later. INSL3 acts through a unique G-protein-coupled receptor, called RelaXin-like Family Peptide receptor 2 (RXFP2), which is expressed by the mesenchymal cells of the gubernacular ligament linking the testes to the inguinal wall. The role of INSL3 in the male fetus is to cause a thickening of the gubernaculum which then retains the testes in the inguinal region, while the remainder of the abdominal organs grow away in an antero-dorsal direction. This represents the first phase of testis descent and is followed later in pregnancy by the second inguino-scrotal phase whereby the testes pass into the scrotum through the inguinal canal. INSL3 acts as a significant biomarker for Leydig cell differentiation in the fetus and may be reduced by maternal exposure to endocrine disrupting chemicals, such as xenoestrogens or phthalates, leading to cryptorchidism. INSL3 may have other roles within the fetus, but as a Leydig cell biomarker its reduction acts also as a surrogate for anti-androgen action.

Morphologic alterations of the genital mesentery implicated in testis non-descent in rats prenatally exposed to flutamide.

BACKGROUND: There is an endless debate on whether androgens mediate testis descent through developmental changes in the gubernacular or the cranial suspensory ligament. OBJECTIVE: To investigate the relation of any possible morphologic changes in the genital mesentery, that is, the system of genital peritoneal folds including the gubernacular and cranial suspensory ligaments, with the event of testis non-descent in rats prenatally exposed to the antiandrogen flutamide. MATERIALS AND METHODS: Time-pregnant Sprague Dawley rats received flutamide (100 mg/kg/d) or vehicle subcutaneously on gestational days 16-17. Flutamide-treated male offspring (n = 67), and vehicle-treated male (n = 34) and female (n = 28) offspring were surgically explored under microscope on postnatal day 50. Testicular position was examined bilaterally. Dimensions of genital mesentery parts were also assessed bilaterally. Association of flutamide-induced morphologic changes with descended (n = 61) and undescended (n = 50; 33 cryptorchid and 17 ectopic) testes was investigated with logistic regression analysis. RESULTS: The male genital mesentery comprised a cranial and a caudal fold converging on the vas deferens. Flutamide resulted in enlarged cranial and reduced caudal folds. Of all flutamide-induced alterations, the increased length of the posterior fixation of the cranial fold and the decreased length of the gubernacular ligament of the caudal fold were found to independently increase the odds of testis non-descent. Testicular ectopy, unlike cryptorchidism, was associated with a short gubernacular ligament only. The female genital mesentery consisted of a cranial fold only. CONCLUSION: Our findings showed a combined contribution of both cranial and caudal folds of the genital mesentery to testis non-descent, through an abnormally long mesentery root and an abnormally short gubernacular ligament, respectively. Inhibition of male-specific development of the genital mesentery with flutamide did not result in a feminized architecture.

Physiology and evolution of the INSL3/RXFP2 hormone/receptor system in higher vertebrates.

Although the insulin-like peptide hormone INSL3 and its cognate receptor RXFP2 (relaxin-family peptide receptor 2) have existed throughout chordate evolution, their physiological diversification appears to be linked closely with mammalian emergence and radiation. In contrast, they have been lost in birds and reptiles. Both hormone and receptor are expressed from autosomal genes which have maintained their synteny across vertebrate evolution. Whereas the INSL3 gene comprises only two exons closely linked to the JAK3 gene, RXFP2 is normally encoded by 18 exons. Both genes, however, are subject to alternative splicing to yield a variety of possibly inactive or antagonistic molecules. In mammals, the INSL3-RXFP2 dyad has maintained a probably primitive association with gametogenesis, seen also in fish, whereby INSL3 promotes the survival, growth and differentiation of male germ cells in the testis and follicle development in the ovary. In addition, however, the INSL3/RXFP2 system has adopted a typical 'neohormone' profile, essential for the promotion of internal fertilisation and viviparity; fetal INSL3 is essential for the first phase of testicular descent into a scrotum, and also appears to be associated with male phenotype, in particular horn and skeletal growth. Circulating INSL3 is produced exclusively by the mature testicular Leydig cells in male mammals and acts as a potent biomarker for testis development during fetal and pubertal development as well as in ageing. As such it can be used also to monitor seasonally breeding animals as well as to investigate environmental or lifestyle conditions affecting development. Nevertheless, most information about INSL3 and RXFP2 comes from a very limited selection of species; it will be especially useful to gain further information from a more diverse range of animals, especially those whose evolution has led them to express unusual reproductive phenotypes.

Sperm DNA damage measured by sperm chromatin structure assay in men with a history of undescended testes.

The aim of this study was to compare sperm DNA damage between men with a history of congenital undescended testis (UDT) and men with a history of acquired UDT. A long-term follow-up study of men with previous UDT was performed. Fifty men with congenital UDT who had undergone orchiopexy at childhood age, 49 men with acquired UDT after a 'wait-and-see'-protocol (e.g. awaiting spontaneous descent until puberty and perform an orchiopexy in case of non-decent), and 22 healthy proven fertile men were included. The DNA fragmentation index (DFI) using sperm chromatin structure assay (SCSA) was used to express the level of sperm DNA damage. Decreased fertility potential was considered if DFI was above 30%. Sperm DNA damage was not statistically different between cases of congenital and acquired UDT. DFI was significantly more often >30% in the complete group of men with congenital UDT (9/50; 18%) and in the subgroup with bilateral congenital UDT (3/7; 43%) in comparison with the controls (none) (p-value 0.049 and 0.01, respectively). Age at orchiopexy in congenital UDT had no statistical effect on DNA damage. In men with acquired UDT, DFI did not statistically differ between those having undergone orchiopexy and those experiencing spontaneous descent. This study supports the hypothesis that UDT is a spectrum representing both congenital UDT and acquired UDT. Sperm DNA damage at adult age is not influenced by age at orchiopexy in congenital UDT cases and by orchiopexy or spontaneous descent in acquired UDT cases.