Electron microscopy of isolated rat testicular cells grown in vitro.
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Biomedical subjects
Publications and source records attributed to O Vilar.
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Adult male rats were treated with prostaglandin E2 (100 microgram/100 g body wt) during 60 days. Neither morphological alterations in genital organs nor modification in epididymal contractility were observed. Nevertheless, a significant diminution of fertility was registered. This was recovered after 30 days following the cessation of treatment. Since sperm maturation occurs in the epididymis, it is postulated that the decrease of fertility in our experiments was due to an incomplete maturation of germinal cells, induced by a reduction in the time taken by spermatozoa in passing through the duct.
The Bucher's method was applied to establish normal values of ATP in the spermatozoa of fertile men. It was shown that the normal value is 6.5 +/- 0.34 S.E. microgram per 10(8) spermatozoa. Pools of spermatozoa showed that incubation in seminal plasma at room temperature produced a progressive loss of ATP in the gametes.
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To study the real phagocytic capacity of Sertoli cells the following experiments were carried out: carbon colloidal solution or thorium dioxide of particle size 0.01--0.03 micrometer as 2% solution were injected in rat testes at a speed of 50 microliter per minute by means of a infusion pump. The site of injection was made either in the intertubular species under the albuginea or in the lumen of seminiferous tubules using glass 120 micrometer-gauge needles connected to the pump syringe. 30 to 240 minutes after the injection the tissues were fixed by perfusion of 2% glutaraldehyde and embedded for EM. It was observed: 1) particles injected into the intertubular species were not able to pass through the tubular wall and were either trapped by the connective tissue macrophages or drained by the lymphatic vessels; 2) intraluminal particles were not phagocyted by the Sertoli cells and remained in the lumen. The behavior of the Sertoli cells did not match that of phagocytes since they did not show their devouring capacity for the inert particles. These results are consistent with the interpretation that the current disposal of either debris by Sertoli cells depends on autophagic dissolution rather than n macrophagic capacity.
The spontaneous contractility of the pelvic urethra and seminal vesicles in the rat was recorded in vivo and the effects of the electrical stimulation as well as the neurohypophysial hormones on it were studied. The mean amplitude of pelvic urethra contractions was 3.2 +/- 0.9 cm H2O and the mean frequency was 4.1 +/- 0.8 contractions/min. The seminal vesicles contractions exhibited a mean amplitude of 1.6 +/- 0.7 cm H2O and a mean frequency of 2.6 +/- 0.8 contractions/min. Each electrical stimulation produced a sudden and vigorous contraction in pelvic urethra as well as in seminal vesicles. Oxytocin and vasopressin did not change neither the amplitude nor the frequency in the spontaneous activity of these organs.
The aim of the present study was to ascertain the effect of methyl xanthines upon the spermatozoa ATP concentration in normal semen in vitro. 26 normal semen were studied. The specimens were diluted after liquefaction with equal volume of Lopata's buffer or Lopata's buffer plus 1 or 6 mM either of caffeine or pentoxifylline. The samples of semen were incubated at room temperature during 90, 180 and 240 minutes before motility and ATP determination by the firefly luciferin-luciferase method. Significant variation was observed in sperm motility nevertheless variations in the ATP concentration was not induced by any of the methyl xanthines we used.
The effects of bilateral gonadectomy, testosterone replacement and cyproterone acetate treatment on seminal vesicle contractions were studied in rats in order to contribute new data on the role of androgens in the spontaneous motility of the seminal vesicle. In gonadectomized-testosterone treated rats the glands exhibited a spontaneous rhythmic activity similar to that of entire animals. The seminal vesicles of gonadectomized as well as of cyproterone treated rats also contracted spontaneously but the contractions showed a smaller amplitude. Such decrease could be related with the well-known atrophy and reduction in the number of smooth muscle cells occurring in seminal vesicles after castration. The results reported in this paper suggest that androgens are not required for the normal spontaneous activity of seminal vesicles.
GPC was studied in the seminal plasma of 35 normozoospermic men, 34 cases of azoospermia due to bilateral deferent obstruction, 34 cases of azoospermia due to bilateral ejaculatory duct obstruction, 10 vasectomized patients, 6 vasectomized patients after vasovasostomy and 118 cases of spermatogenetic arrest without obstruction of the seminal ducts. Values of GPC in azoospermia due to deferent or ejaculatory ducts obstruction and to vasectomy are significantly lower than in normozoospermic subjects (p less than 0.001). Levels of GPC increased (p less than 0.05) following vasovasostomy. Although GPC values in secretory azoospermia were higher than those in cases of duct obstruction, they were still lower than in normozoospermic (p less than 0.001). The most likely source of GPC is the epididymis. These results support the assumption that GPC originates mainly in the epididymis. The absence of germinal cells in the epididymis could explain the decreased levels of GPC in azoospermia due to arrest of spermatogenesis.
The aim of the present paper is deleted to determine ATP activity in the human seminal cells by bioluminescence and to investigate the relationship between these values and the number of round cells either peroxidase positive or peroxidase negative. We studied 244 untreated men, divided into six groups. Our mean value of ATP activity in normal semen was 20.02 +/- 0.65 n moles per 10(8) spermatozoa. In asthenozoospermic or oligoasthenozoospermic patients with less than 1,000,000 round cells per ml, the concentration of ATP was significantly lower than normal (P less than 0.001) or (P less than 0.05). Semen with normal characteristics or samples from asthenozoospermic or oligoasthenozoospermic patients but with more than 1,000,000 round cells, per ml and with predominance of peroxidase negative cells, had an ATP concentration higher than normal subjects (P less than 0.001). We believe that the knowledge of the quantity and quality of round cells of the ejaculate is important to interpret the seminal values of ATP.
Fertility of men depends on the quality of semen. The aim of the present paper is to determine both the acrosin activity by radioimmunoassay and ATP concentration by bioluminescence in human spermatozoa, and evaluate these results in those samples with normal or low sperm penetration according to SPA test. Ejaculates obtained from 42 untreated men, were studied one hour after the obtention. These materials were divided into two groups:20 human semen samples with "in vitro" potentiality to penetrate zone-free hamster ova, between 15% to 98% and 22 human semen with SPA test between 0% to 14%. When we compare the group with normal penetration response vs that group with low or absent penetration ones, a significant decrease of ATP and acrosin concentrations was observed (P less than 0.001). Nevertheless no significant difference was observed in relation with percentage of motility, volume (ml), sperm concentration (10(6)/ml), percent of quick progressive spermatozoa and number of gametes capable of migrating into the medium layer (10(6)), between the group with low or absent penetration test against that one with normal zona-free hamster egg test.
Salivary sialic acid concentration was studied daily in 20 fertile women during the menstrual cycle. The Warren method was used and plasma LH and BBT were also determined. The sialic acid concentrations in saliva throughout the cycle showed a peak 4 or 5 days before the LH preovulation surge. This maximal peak showed a mean of 10.89 +/- 1.11 mg%. This method may be useful for predicting the pre- and postovulatory fertile period.
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