Browsing by Subject "Hoden"

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Geschlechtsspezifische Unterschiede in der Fötalentwicklung beim Schwein
(2007) Häußler, Susanne; Claus, Rolf
Basic mechanisms of sexual differentiation in higher mammals are well established. The development of the testes is controlled by genetic mechanisms and initiated by the Y-chromosome. Further differentiation of the ?Anlagen? is performed by the presence of testicular androgens but requires no specific signal in females (basic femaleness). Speculation exists during the fetal development of pigs, because androgens are also measurable in female fetuses. In addition, the male gonad is able to synthesize remarkably high levels of estrogens. The aim of the present study was to follow up concentrations of steroids in peripheral plasma throughout fetal development, starting with week 6, and in particular to analyze changes in testicular cell populations (spermatogonia, Leydig cells) and to correlate them with testicular androgens, estrogens, 19-nortestosterone and cortisol. The expression of steroid converting enzymes such as 11beta-HSD 2 and aromatase as well as receptors were determined by immunocytochemistry and quantitative PCR. Altogether each of the stages of gestation (weeks 6, 10, 13, and 15 of pregnancy) was represented by 4 sows, so that a total of 158 fetuses were collected. Testicular steroid synthesis (testosterone, estradiol) could be demonstrated as early as week 6, but was independant of gonadotropine. 19-nortestosterone, which is formed during estrogen synthesis, was detected in amniotic fluid using a new established enzymeimmunoassay in this study. Aromatase activity clearly correlated with a wave-like pattern of cell development. Therefore the activity was elevated both during an alternating rise of Leydig cells or spermatogonia mitosis. During the rise of spermatogonia development Leydig cells remained quiescent and during the Leydig cell mitosis spermatogonia remained inactive. An increased aromatase activity was observed both during the rise of spermatogonia and Leydig cell mitosis, and in consequence an elevated concentration of estradiol was found. But during an increased Leydig cell formation aromatase expression and thus estradiol synthesis was taken over by spermatogonia. It is therefore reasonable to resume that estrogens are important mitogenic signals as it was also found earlier in mature boars. The expression of glucocorticoid receptors by spermatogonia could be demonstrated for the first time in fetal pig testes. As also shown for other tissues, its likely role in testes is the differentiation of new cells. This important role also explains the expression of the enzyme 11beta-HSD 2 both by Leydig cells and spermatogonia. This enzyme is a well known fine-tuning mechanism which indicates cortisol and thus the ligand for the glucocorticoid receptor. Its expression in parallel to the rise of estrogens suggests a dependancy on estrogens. Investigating this was, however, not the topic of the present study. The demonstration of androgens both in blood plasma and amniotic fluid in female fetuses seems to contradict the principle of basic femaleness. It was shown however, that concentrations of testosterone in males increase up to 2.01 ng/ml plasma during the main period of sexual differentiation whereas female levels remain at 0.2 ng/ml so that it is simply the concentration which decides where the male differentiation does occur. It can not be ruled out, however, that low concentrations in female fetuses may have an effect on follicular differentiation, as was also demonstrated in mature sows. At the same time androgens could have a mitotic effect caused by insulin-like growth factors (IGF I + II). Thus the present investigation was able to clarify of several new mechanisms and basic data of fetal pig development. Further confirmation of the mechanisms suspected in this study may be served by an aromatase inhibitor.
Steuerungsmechanismen der Spermatogenese beim Eber: Auswirkungen einer GnRH-Immunisierung und anschließender Estradiolinfusion
(2005) Wagner, Anna; Claus, Rolf
Whereas the regulation of spermatogenesis is well known in various species, specific differences, however, exist for the boar, which require species-specific investigation. High concentrations of estrogens are synthesized in the boar testis. It is known, that they play a synergistic role for male behaviour and function of accessory sex glands. Additionally, estrogens in the ejaculate reach the female genital tract and contribute to sperm transport and ovulation in the sow. A possible involvement of estrogens in spermatogenesis, however, was not investigated so far. Similarly, high concentrations of glucocorticoids occur in the tubuli of mature boars. Their regulatory function in spermatogenesis so far is unknown. For clarification, boars were actively immunized against GnRH. GnRH stimulates gonadotropin-release from the pituitary. The evaluation of hormone concentrations in blood plasma after GnRH immunization showed unexpectedly, that the gonadotropin FSH is not influenced, whereas LH- and in consequence steroid- (androgen and estrogen) concentrations were abolished. This offers the possiblity to characterize the selecitve function of estrogens in the presence of FSH. Three groups were evaluated: intact boars (n=5), immunized boars (n=5), 17ß-estradiol infused, immunized boars (n=6). All animals were fitted with indwelling cephalic vein catheters on both sides. One of them was reserved for infusion, the other one for uncontaminated blood sampling. All animals were infused with physiological saline over 7 weeks. For the 17ßEstradiol infused, immunized boars, a defined concentration of estradiol was added to the solution. Blood samples were collected over the whole 7-week period for LH, FSH, testosterone and estradiol. Blood samples were later analysed by established radioimmunological procedures. At the end of the 7-week period, all animals were killed. Tissue samples were fixed for histological examination. Morphological criteria included e.g. tubular diameter, seminiferous epithelial height and number of Leydig cells. Imunocytochemical evaluation was performed for mitosis (Ki-67), apoptosis (TUNEL) and the glucocorticoid receptor. The number of germ cell in the tubuli was reduced by 60% in immunized boars compared to the intact controls. These changes were explained by a more pronounced expression of the glucocorticoid receptor in spermatogonia, followed by the appearance of apoptosis in spermatogonia and primary spermatocytes. In consequence these findings demonstrate for the first time an involvment of glucocorticoids in the quantitative regulation of spermatogenesis. They counteract tubular mitosis by reducing the number of germ cells by apoptosis. In the 17ßestradiol infused group of immunized boars, the infusion led to the restoration of physiological estradiol concentrations (230 pg/ml) in peripheral blood plasma. As expected, androgen concentrations were minimal, so that the specific role of estrogens for spermatogenesis could be clarified. Data from the histological evaluation showed, that the expression of the estrogen receptor alpha occured in spermatogonia, selectively in the mitotic active stage of the seminiferous epithelial cycle leading to a 41-50% rise of germ cell numbers compared to immunized boars. The apoptotic rate in the tubuli of the estradiol infused boars however was not changed compared to immunized boars, so that physiological sperm numbers as they are characteristic for intact boars, were not reached. This study shows for the first time that both glucocorticoids and estrogens play an essential role for spermatogenesis in the boar. They are involved in the regulation of sperm yield by influencing the mitosis-apoptosis equilibrium in the tubuli of boars.