Effects of neonatal estrogenization on germ cell development.
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Biomedical subjects
Publications and source records attributed to F Gaytan.
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The effect of a new commercial interferon (Fiblaferon L) on the human immune system was studied. Different experiments were performed on human peripheral blood in vitro, in order to observe: (a) the phagocytic capacity of polymorphonuclear leukocytes (PMNL) and monocytes; (b) monocyte adherence index, and (c) percentage of antibody-dependent cellular cytotoxicity (ADCC) against chicken red blood cells (CRBC). Fiblaferon L enhanced all these functions.
The appearance of cellular associations between macrophages and lymphocytes--which we have denominated macrophage-lymphocyte rosettes--and their kinetic formation in the presence of phytohemagglutinin (PHA) have been studied in B10 A (4R) mice. The greatest number of macrophage-lymphocyte rosettes was found from 6 to 12 hours after incubation with PHA. During this time, 42.38 +/- 10.70 of the total number of macrophages had lymphocytes attached to their membranes. This percentage decreased to 17.33 +/- 2.07% after 24 hours. The activation of macrophages after PHA treatment was tested by the phagocytic capacity of these cells. This activity increased significantly 24 hours after incubation. In our assay, an increase in the appearance of multinucleated giant cells when compared to controls was also observed. When the macrophages were lymphocyte depleted, the appearance of the multinucleated giant cells was significantly lower. The kinetics for these formations are also discussed.
An ultrastructural and morphometric study of the testes in 15-, 22-, 45-, and 90-day-old neonatally estrogenized rats was performed. At 45 days of age, the Sertoli cells appeared immature in estrogenized rats, whereas they were fully mature in the controls. This finding might be related to a deficiency in gonadotropins and androgens during the postnatal period. In 90-day-old estrogenized rats, however, Sertoli cell maturation had occurred, which might be attributed to a recovery of hormone levels. Cytoplasmic alterations, however, such as vacuolation, were present at this age. The morphometric study revealed decreased testicular and tubular volumes as well as decreased mean tubular diameters in the estrogenized animals. In contrast, the absolute tubular length increased more in these animals than in the controls during the period from 15 to 90 days of age. This lengthening process might be related to the large number of hypercurved tubules in the estrogenized rats.
Mast cells in the testis of control adult rats were found almost exclusively around subcapsular blood vessels. Discrete mast cells were distributed throughout the stroma of the epididymis and sex accessory glands. In neonatally estrogen-treated rats, a greater number of mast cells was present in the testicular interstitium, whereas no significant increase in the number of mast cells per square millimeter of stroma was found for the epididymis and sex accessory glands, despite stromal proliferation. On the other hand, androgen-treated rats did not have increased mast cell numbers in any organ. These results indicate that the increase in mast cell numbers was estrogen-dependent, specifically related to the testis and did not seem to be a consequence of the increase in the connective interstitial tissue.
The presence of cells bearing solitary cilia has been studied in the testes of normal and neonatally estrogenized rats. Peritubular myoid cells and undifferentiated interstitial cells showed a higher frequency of cilia in estrogenized animals than in control ones, from 15 to 45 days of age, the differences being slighter at 90 days of age.
The response of testicular macrophages to massive Leydig cell death was studied by the administration of the specific Leydig cell cytotoxic ethylene dimethane sulphonate (EDS) to sham-operated (SO), short-term (STHX), and long-term (LTHX) hypophysectomized rats. EDS-killed Leydig cells showed the morphological features of the programmed cell death or apoptosis. A 2-fold increase in the number of macrophages was found on days 1-2 after treatment in both SO and STHX rats, and dead Leydig cells were completely eliminated by day 3 after treatment. Otherwise, in LTHX rats, there was a delay in the increase in the number of macrophages, and EDS-killed Leydig cells remained in the testicular interstitium for several days. These results indicate that the phagocytic capacity of the macrophage population was diminished in hypophysectomized rats, and particularly after long-term hypophysectomy.
We have studied the response of atrophic Leydig cells to gonadotrophin replacement in young hypophysectomized (HX) and GnRH antagonist (GnRH-ANT)-treated rats. Hypophysectomy was performed at 28 days of age. Age-matched rats were treated with GnRH-ANT from 28 to 51 days of age. From 45 to 51 days of age, animals were injected with 5 IU recFSH, 10 IU hCG or vehicle. Body and testicular weights, as well as the diameter of the seminiferous tubules were significantly higher in GnRH-ANT-treated than in HX rats. Both recombinant FSH and hCG treatments induced a similar increase in testicular weight and tubule diameter in HX and GnRH-ANT-treated rats. However, hCG treatment induced a significantly higher increase in Leydig cell size in HX (3.2-fold) than in GnRH-ANT-treated (1.4-fold) rats. These results suggest that the response of atrophic Leydig cells to gonadotrophin supplementation was partially inhibited in the presence of GnRH antagonist, whereas Sertoli cell-mediated responses seem not to be affected.
The proliferation and differentiation of mast cells and Leydig cells were studied in adult sham operated or hypophysectomized rats after the administration of ethylene dimethane sulphonate (EDS) and in prepubertal rats after neonatal treatment with a gonadotropin-releasing hormone (GnRH) antagonist (Organon 30276; Oss, The Netherlands). After treatment with EDS, two proliferative waves were found. On day 3, several interstitial cell types proliferated, whereas mitotic cells corresponded to differentiating Leydig cells and mast cells around day 20. Differentiating Leydig cells showed a higher mitotic index than that of differentiating mast cells. Hypophysectomized animals showed high mitotic activity 3 days after treatment, but 21 days after treatment differentiating Leydig cells were absent and proliferative activity was reduced. The number of mast cells increased from day 15 to day 30 in EDS-treated rats and from day 15 to day 50 in hypophysectomized, EDS-treated rats. GnRH antagonist-treated rats showed poorly differentiated Leydig cells and abundant mitotic figures on day 23. Proliferation and differentiation of Leydig cells occurred concomitantly with the proliferation and differentiation of mast cells between 23 and 30 days of age. These results suggest that Leydig cells and mast cells in the rat testis share some common regulatory factors.
In order to correlate the date of balano-preputial separation (BPS) with the testicular development, twenty four rats were selected from an experimental design focused on the effects of pituitary grafts on puberty. Animals that presented BPS at an early age, showed a smaller volume of the seminiferous epithelium and a lower spermatogenic level than that presented BPS at a more advanced age. These data indicate that the advancement in BPS induced by pituitary grafts was not in keeping with an equivalent enhancement of testicular development.
Testicular macrophages were selectively depleted in the right testes of adult rats by an intratesticular injection of dichloromethylene diphosphonate-containing liposomes (Cl2MDP-lp), whereas the left testes were injected with 0.9% NaCl and served as control. Before or after Leydig cell destruction with ethylene dimethane sulfonate (EDS), treatment with Cl2MDP-lp/NaCl was given at different times to study the requirements of macrophages in the different stages of Leydig cell regeneration. On day 30 after EDS treatment, new Leydig cells were abundant in the left, macrophage-containing testes. However, in the right, macrophage-depleted testes, the number of Leydig cells was related to the time elapsed between EDS treatment and macrophage depletion. When macrophages were depleted on day 10 before or on days 4 or 10 after EDS treatment, new Leydig cells were nearly absent at 30 days. However, when macrophages were depleted on days 16 or 22 after EDS treatment, Leydig cells were found at 30 days, but their numbers were equivalent to the number of Leydig cells that were already present in EDS-treated animals at the time the macrophages were depleted. These results indicate that macrophages are needed for the differentiation of Leydig cells from mesenchymal precursors, as well as for the proliferative activity of the newly formed Leydig cells, possibly through the secretion of essential growth factors.