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Biomedical subjects

S Fasano

Publications and source records attributed to S Fasano.

At least 37 records · Page 2Linked to original sources
PubMed1994

Regeneration of the testicular interstitial compartment after ethane dimethane sulfonate treatment in the hypophysectomized frog Rana esculenta: independence of pituitary control.

The effects of ethane dimethane sulfonate (EDS) on the testes of hypophysectomized frogs (Rana esculenta) were investigated by light and electron microscopy. Initial signs of interstitial cell damage were observed in EDS- and EDS plus pituitary homogenate (PH)-treated animals 5 days after a single injection of EDS (100 mg/Kg body weight). The germinal compartment in these two groups appeared disorganized adjacent to the damaged interstitial tissue only in the EDS-treated animals, and by Day 8, spermatogenesis seemed to be affected in the EDS + PH-treated frogs in which Leydig cells had disappeared in some areas. On Day 28, regeneration of the interstitial tissue was complete and spermatogenesis was restored to normal. These data suggest that, in hypophysectomized frogs, the regeneration of the interstitial compartment is independent of pituitary activity and that the lack of interstitial cells activates the production of local factors responsible for the differentiation and proliferation of new Leydig cells. It is concluded that in addition to gonadotropins, the intratesticular environment is fundamental in the maintenance and regulation of testicular structure and function.

Androgens↗
PubMed1994

Detection and localization of gonadotrophin-releasing hormone (GnRH)-like material in the frog, Rana esculenta, ovary.

GnRH-like material has been identified using HPLC followed by RIA in the ovary of Rana esculenta. During the reproductive cycle three immunoreactive GnRH peaks were eluted. One of them coeluted with s-GnRH, the other two forms between GnRH and cII-GnRH. During the recovery phase s-GnRH immunoreactivity disappears. By immunocytochemistry, cII-GnRH immunostaining was localized to granulosa cells while s-GnRH was present in the perinuclear zone of the oocytes.

Animals↗
PubMed1994

Opioid peptides and testicular activity in the lizard Podarcis s. sicula Raf.

In mammals endorphinergic systems have been shown to modulate reproductive processes and beta-endorphin (beta-EP) has been found to influence sexual functions, acting at the hypothalamus-pituitary-gonadal axis level. Using immunocytochemical and in vitro studies, evidence for a diffuse pro-opiomelanocortin-related opioid system in the lizard Podarcis s. sicula was produced. In the testis, beta-EP immunoreactivity showed seasonal variation, being most pronounced in the interstitial cells of sexually quiescent lizards (December). Reverse-phase high-performance liquid chromatography, coupled with radioimmunoassay and immunocytochemistry, showed that beta-EP and acetyl beta-EP increased during December, while their concentrations were low during April, when the highest testicular activity occurred. Using in vivo studies, it was found that naltrexone treatment, blocking pituitary opioid receptor, increased androgen levels in the plasma and in the testis. It was also found with in vitro studies that the endogenous opioid system inhibits gonadotrophin release and therefore androgen production by the testis. The data reported here provide evidence for the physiological role played by opioid peptides at the pituitary level to regulate the seasonal reproductive activity of the lizard Podarcis s. sicula.

Androgens↗
PubMed1993

Two GnRHs fluctuate in correlation with androgen levels in the male frog Rana esculenta.

ChickenII-(cII-) and salmon (s-) GnRH levels have been measured in the male frog Rana esculenta during the annual cycle. The presence of pituitary binding activity for both peptides and plasma androgen levels has been investigated in order to give insight into the significance of the dual control exerted by the GnRH forms present in the R. esculenta brain. ChickenII- and s-GnRHs showed high values during the spring-summer period. Conversely, while cII-GnRH peaked in February, s-GnRH declined slowly from February until May. Plasma androgen levels increased as the peptides decreased during the autumn-winter period. Still high androgen levels (but significantly lower as compared with winter concentrations) were found during spring. Using iodinated cII-GnRH, GnRH binding sites were detected in pituitary preparations when the corresponding peptide concentration decreased in the brain. On the contrary, no binding sites were found using labeled s-GnRH. Our results indicate that cII-GnRH has a hypophysiotropic activity, while the role of s-GnRH needs to be further investigated.

Androgens↗
PubMed1993

Dopamine regulation of testicular activity in intact and hypophysectomized frogs, Rana esculenta.

In intact frogs, both GnRHA and L-dopa were able to increase testicular and plasma androgen levels and to induce spermiation. The dopamine antagonist pimozide inhibited both the effects of L-dopa but not those of GnRHa. Hypophysectomy reduced androgen levels, but spermiation was still induced by both GnRHa and L-dopa, suggesting that these agents can directly influence the testis through a route not involving the pars distalis. Again, pimozide antagonised spermiation induced by L-dopa but not that induced by GnRHa.

Androgens↗
PubMed1993

Opioids and testicular activity in the frog, Rana esculenta.

The presence and activity of brain, pituitary and testicular beta-endorphin (beta-EP)-like material have been studied in the frog, Rana esculenta, using reverse-phase high-pressure liquid chromatography, coupled with radioimmunoassay and immunocytochemistry. In-vivo and in-vitro treatments with naltrexone were carried out to assess the putative physiological activity of opioid peptides. beta-EP(1-31) and (1-27), together with their acetylated forms, have been identified in brain, pituitary and testis. In particular, beta-EP(1-31) concentrations peaked during July in the brain and pituitary, whilst in testes maximum concentrations were found in April and November. beta-EP immunoreactivity was present in the brain within the nucleus preopticus and nucleus infundibularis ventralis while positive fibres in the retrochiasmatic regions projected to the median eminence. In the testis, interstitial cells, canaliculi of the efferent system, spermatogonia and spermatocytes showed positive immunostaining for beta-EP. In intact animals, naltrexone treatment increased plasma and testicular androgen levels and this effect was confirmed in in-vitro incubations of minced testes. Naltrexone also induced a significant increase in germ cell degeneration. Our results indicated that an opioid system modulates the hypothalamus-pituitary-gonadal axis in the frog, Rana esculenta and, for the first time, we have shown that the testicular activity of a non-mammalian species may be regulated by opiates locally.

Animals↗
PubMed1992

Intratesticular control of spermatogenesis in the frog, Rana esculenta.

Adult intact and hypophysectomized (PDX) frogs, Rana esculenta, were treated with a gonadotropin releasing hormone agonist (GnRHA, HOE 766) and/or cyproterone acetate (CPA), the antiandrogen, in order to investigate the regulation of primary spermatogonial (I SPG) multiplication in vertebrates. Treatment with GnRHA (injections containing 900 ng administered for 12 days on alternate days) caused a significant increase of the mitotic index (MI) of I SPG in PDX animals and a further MI increase of SPG was observed when 0.66 mg CPA was given concomitantly with GnRHA. The treatment with 0.66 mg CPA in combination with GnRHA also increased secondary spermatocyte (II SPC) appearance. Moreover, number of nests containing spermatids (SPT) decreased as CPA, in combination with GnRHA, was administered in increasing doses (0.33 and 0.66 mg/injection). Intact animals treated with CPA (0.66 mg/injection) showed a time-dependent I SPG multiplication increase which reached highest values after 28 days. Secondary SPC also proliferated until day 28; meanwhile the number of nests containing SPT decreased. Neither testosterone nor R5020 (a progestin which is not converted to androgens) modified the basal and GnRHA-induced spermatogonial proliferation. These results confirm that in the frog, Rana esculenta, spermatid formation is impaired by CPA treatment and that I SPG multiplication is enhanced by a direct effect of GnRHA; moreover, we suggest that the absence of spermatids constitutes a signal promoting spermatogonial proliferation.

Animals↗
PubMed1992

Effects of gonadotropin-releasing hormone variants on plasma and testicular androgen levels in intact and hypophysectomized male frogs, Rana esculenta.

The effects of vertebrate gonadotropin-releasing hormone (GnRH) variants on plasma and testicular androgen level in intact and hypophysectomized (PDX) male frogs, Rana esculenta, have been investigated. In intact animals, mammalian (m)-GnRH, m-GnRH analog (buserelin), salmon (s)-GnRH, chicken (c) I-GnRH, cII-GnRH, D-Arg6-cII-GnRH (cII-GnRHA), and lamprey (l)-GnRH (1.5 micrograms and 6 micrograms, total dose given on alternate days for 5 days) were able to enhance androgen production showing that specificity of pituitary responsiveness to GnRH variants appears to be low. Chicken II-GnRH was more effective than s-GnRH in eliciting testicular and circulatory androgen level increase. Moreover, in animals treated with 6 micrograms of cII-GnRH and s-GnRH in combination, androgens decreased as compared with animal treated with cII-GnRH only, suggesting that GnRH receptors bind preferentially the s-GnRH form. In PDX animals, buserelin (1.5 and 6 micrograms), cII-GnRH, and its analog (6 micrograms) were able to increase plasma androgen levels whereas testis androgen concentrations were increased by cII-GnRH (1.5 and 6 micrograms), D-Arg6-cII-GnRHA, and buserelin (6 micrograms). Since androgen production in PDX animals is influenced especially by peptides sharing cII-GnRH structure, it is suggested that a testicular cII-GnRH-like material play a role as local modulator of the gonadal activity in Rana esculenta.

Amino Acid Sequence↗
PubMed1992

Plasma and follicular tissue steroid levels in the elasmobranch fish, Torpedo marmorata.

Steroid concentrations in plasma and follicular tissues (theca plus granulosa layers) were determined by radioimmunoassay in the aplacental viviparous ray, Torpedo marmorata, during various stages of the reproductive cycle. Steroids in the uterine fluid of pregnant animals and in preovulatory atretic follicles were also measured. In the follicular tissue of cyclic animals, levels of progesterone were always lower than those of estradiol-17 beta and androgens (testosterone plus 5 alpha-dihydrotestosterone). Estradiol-17 beta and androgen levels increased as the animals approached the ultimate maturational stage before ovulation. Androgens were not detectable in plasma, while estradiol-17 beta increased dramatically before ovulation. In pregnant animals, only small ovarian follicles (less than 5 mm in diameter) were observed, and these had hormone concentrations that were similar to those of the small follicles of cyclic animals. Progesterone was the only steroid detected in the uterine fluid of pregnant animals. In completely sclerotic atretic follicles of pregnant animals, steroids were not detected. Progesterone was the main hormone in atretic follicles undergoing yolk resorption. This suggests that the latter may contribute to the elevated plasma progesterone concentrations of pregnant animals.

Animals↗
PubMed1992

Immunoreactive Met-enkephalin-like material in the testis of Rana esculenta: identification and localization.

Methionine-enkephalin (Met-Enk) has been detected in the testis of the frog, Rana esculenta, using a reverse-phase high-performance liquid chromatography system coupled with a specific radioimmunoassay. By means of immunocytochemical techniques Met-Enk positive cells have been localized in interstitial and germinal compartments. Particularly, spermatogonia, spermatocytes, and spermatozoa were stained in seminiferous tubules, and numerous interstitial cells showed strong cytoplasmic immunoreactivity in summer animals. Variations in the concentration of Met-Enk immunoreactive material occurred during the annual cycle. Our data show that Met-Enk is present in testes of nonmammalian vertebrate species. These results suggest that autocrine and/or paracrine mechanisms may regulate testicular activity in amphibians.

Amino Acid Sequence↗
PubMed1992

Ovarian opioids and the reproductive cycle of the frog Rana esculenta.

In mammals, proopiomelanocortin-related peptides are involved in reproductive processes both at the hypothalamo-pituitary and ovarian levels. Using immunocytochemical, biochemical and physiological "in vitro" studies, we provide here evidence for a diffuse POMC-related opioid system in the frog Rana esculenta. Ovarian beta-endorphin (beta-EP) is expressed in thecal cells and changes during the reproductive cycle in an inverse relationship with follicular development. Seasonal changes in the ovary are different to those in the brain or in the pituitary. The ratio of acetylated vs native beta-EP in the ovary also changes over the reproductive period, affecting the biological activity of the peptide. During both the reproductive spring period and the summer post-reproductive phase pMol amounts of beta-EP stimulate follicular androgen secretion in vitro, in a naloxone-reversible way. In either period, an inhibition of estradiol, possibly mediated via other factors, is the result of opioid action. In conclusion, these data demonstrate for the first time the widespread presence of beta-EP-related peptides in the frog Rana esculenta. Both immunocytochemical and biochemical evidence, as well as in vitro responses, support a physiological role for beta-EP in ovarian seasonality during the reproductive cycle of this amphibian.

Amino Acid Sequence↗
PubMed1992

Resumption of testicular activity in Gobius paganellus after administration of ethane 1,2-dimethane sulfonate (EDS).

1. The effect of a single injection of ethane-1,2-dimethane sulfonate (EDS) was studied in the teleost fish, Gobius paganellus in two different periods of the year. 2. During June EDS did not induce any change, while during December the drug was highly effective in promoting testicular activity. 3. Nucleus/cytoplasm ratio of interstitial cells strongly decreased concomitantly with the detection of high testicular androgen levels. 4. The germinal compartment was well developed showing the appearance of all spermatogenic stages and the cavity of lobular compartments filled of spermatozoa. 5. Our data are the first evidence of a stimulatory activity of EDS on testes of a vertebrate species.

Androgens↗
PubMed1991

Sites of action of local estradiol feedback mechanism in the frog (Rana esculenta) testis.

The direct effect of estradiol on testicular androgen biosynthesis was studied in the frog, Rana esculenta, measuring progesterone, 17 alpha-OH-progesterone, androstenedione, and androgens (T + DHT) in supernatants and testes incubated (6 hr, 15 degrees) with estradiol alone (10(-6) M) or in combination with crude pituitary homogenate (1 pituitary equivalent/tube). Estradiol, either alone or in combination with pituitary homogenate, induced decreases of 17 alpha-OH-progesterone, androstenedione, and androgens but was ineffective in modulating progesterone levels. Pituitary homogenate was effective in inducing a significant increase of androstenedione and androgens but was ineffective in modulating both progesterone and 17 alpha-OH-progesterone production. It is concluded that estradiol acts by decreasing the activity of steroidogenic enzymes starting from 17 alpha-hydroxylase, while pituitary homogenate does not affect the 17 alpha-hydroxylase activity, but it acts starting from 17,20-lyase.

17-alpha-Hydroxyprogesterone↗
PubMed1990

Characterization of gonadotropin-releasing hormone (GnRH) binding sites in the pituitary and testis of the frog, Rana esculenta.

Frog, Rana esculenta, pituitary and testis gonadotropin-releasing hormone (GnRH) receptors were characterized by using 125I-chicken IIGnRH (cIIGnRH) as radiolabeled ligand. At 4 C equilibrium binding of 125I-cIIGnRH to pituitary homogenates was achieved after 90 min of incubation; binding of 125I-cIIGnRH to testis membrane fractions reached its maximum at 60 min of incubation. Binding of the radioligand was a function of tissue concentration, with a positive correlation over the range 0.5-2 tissue equivalents per tube. One pituitary and one testis per tube were used as standard experimental condition. Incubation of the pituitary homogenate with increasing concentrations of 125I-cIIGnRH indicated saturable binding at radioligand concentrations of 1 nM and above while for the testis membrane preparation saturation was achieved using 5 nM 125I-cIIGnRH. The binding of 125I-cIIGnRH was found to be reversible after addition of the cold analog and the displacement curves could be resolved into one linear component for both tissues. Scatchard analysis suggested the presence of one class of binding sites for both pituitary and testis (Pituitary: Kd = 1.25 +/- 0.14 nM and Bmax = 8.55 +/- 2.72 fmol/mg protein; testis: Kd = 2.23 +/- 0.89 nM and Bmax = 26.48 +/- 7.39 fmol/mg protein). Buserelin displaced the labeled 125I-cIIGnRH with a lower IC50 as compared with cIIGnRH cold standard, while Arg-vasopressin (AVP) was completely ineffective, confirming the specificity of binding.

Amino Acid Sequence↗
PubMed1990

Morphological and hormonal changes in the frog, Rana esculenta, testis after administration of ethane dimethane sulfonate.

Apart from mice, in rodents ethane dimethane sulfonate (EDS) selectively destroys Leydig cells. This has been indicated as a new method for the study of seminiferous interstitial compartment interaction. No information on the possible destruction and repopulation of Leydig cells exists in lower vertebrates. This study deals with EDS effects in the frog, Rana esculenta. Animals received a single intraperitonial dose (100 mg/kg body wt) and were sacrificed at 0, 12, and 24 hr and 3, 4, 7, 14, and 28 days postinjection. Androgens (testosterone + DHT) were measured in plasma and right testes. Moreover, left testes were fixed and examined for histological observation. Plasma androgen levels were extremely low on Day 4 after EDS treatment and remained unchanged thereafter. In testes, androgen levels decreased on Day 4 but increased to control levels on Day 14. Leydig cells were damaged within 3 days post-treatment and were completely destroyed on Days 4 and 5. Germinal compartment damage appeared only where the adjacent interstitial tissue presented complete destruction. Pale primary spermatogonia (stem cells) were always present. Testes restored to normal on Day 14 and spermatogenesis resumed to the regenerating interstitial tissue. These results show that regenerating testes in R. esculenta retain androgens and that interstitial-germinal compartment communications may have a role in maintaining spermatogenesis.

Androgens↗
PubMed1990

Indirect evidence for a physiological role exerted by a "testicular gonadotropin-releasing hormone" in the frog, Rana esculenta.

The possible physiological role of a putative testicular gonadotropin-releasing hormone (GnRH)-like material was studied in the frog, Rana esculenta. We have investigated (a) changes of mitotic index (MI) of primary spermatogonia (SPG) in GnRH agonist (GnRH-Ag)-treated testes in vitro; (b) changes of androgen concentrations in testes of intact frogs treated with a GnRH antagonist (GnRH-Ant); (c) variations of mitotic index of primary SPG in intact GnRH-Ant-injected animals and in testes incubated with GnRH-Ant; (d) changes of MI in hypophysectomized (PDX) animals treated with hypophysis (PD) homogenate, and hCG alone or in combination with GnRH-Ant; and (e) changes of androgen concentrations in plasma PDX frogs treated with hCG and hCG plus GnRH-Ant. Our results indicate that while GnRH-Ag induced accumulation of primary SPG mitosis, GnRH-Ant inhibited androgen production and natural occurring mitosis accumulation. Therefore, GnRH-Ant may counteract an endogenous peptide working into the testis.

Androgens↗
PubMed1990

Regulation of the testicular activity in the marine teleost fish, Gobius paganellus.

Seasonal variations of intratesticular steroid hormones (androgens and estradiol-17 beta) and spermatogenic activity have been studied in the marine teleost fish, Gobius paganellus. In addition, in vivo and in vitro experiments have been carried out in order to investigate the control of androgen production by the testis. While estradiol was never detected, androgens were at low values in autumn and reached maximal levels in spring concomitantly with the highest testis weight and the highest efficiency of the spermatogenic wave. In vitro incubations were carried out using ovine luteinizing hormone (oLH) (400, 4000, and 40,000 micrograms/liter; 20 degrees for 6 and 24 hr). The effective dose 40,000 micrograms/liter was used to induce androgen stimulation in both autumn and spring testes. The responsiveness to oLH was enhanced in spring testis. Estradiol and a gonadotropin-releasing hormone analog GnRHA (HOE766) were ineffective in modulating androgen production either alone (1-1000 nmol/liter) or in concert with oLH during short-term incubations. In intact animals, GnRHA elicited, 3 hr after the injection (10 micrograms), a three-fold increase of intratesticular androgen content. In conclusion, we show that the annual androgen profile in G. paganellus parallels the spermatogenic activity and that the androgen production is not affected in these experimental conditions by putative intratesticular factors (e.g., estradiol-17 beta and GnRH-like substances) which, conversely, are effective in inducing androgen changes in several vertebrate species.

Androgens↗