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Biomedical subjects

R Pierantoni

Publications and source records attributed to R Pierantoni.

At least 37 records · Page 2Linked to original sources
PubMed1995

Nature and distribution of gonadotropin-releasing hormone (GnRH) in the brain, and GnRH and GnRH binding activity in serum of the spotted dogfish Scyliorhinus canicula.

The distribution of different molecular forms of gonadotropin-releasing hormone (GnRH) in the brain and serum of the spotted dogfish, Scyliorhinus canicula, was investigated by an indirect immunofluorescence method, using antisera against salmon (s-), chicken-II (cII-) and mammalian (m-) GnRHs, and by reverse-phase high-performance liquid chromatography (RP-HPLC) coupled with radioimmunoassays. Five GnRH molecular forms were demonstrated on the basis of the retention time in the RP-HPLC system. The characteristics of four of these GnRH peptides are consistent with those of m-, cII-, dogfish (df-), and sGnRH. The fifth form appears to be novel. Immunoreactive sGnRH structures were confined to the diencephalon; whereas cIIGnRH and mGnRH were found in the telencephalon and diencephalon. cIIGnRH- and dfGnRH-like molecules were detected in the serum. Moreover, a specific, low-affinity GnRH binding protein (GnRH-BP) was found in the serum of the spotted dogfish. The binding of [125I]sGnRHA to the serum GnRH-BP was dependent on incubation time, equilibrium being reached within 1 hr at 4 degrees; binding was rapid and completely reversible. Scatchard analysis yielded a linear plot with a Kd of 7.9 x 10(-7) M. The presence of a GnRH-BP in spotted dogfish serum suggests a probable action of GnRH via the general circulation.

Animals↗
PubMed1995

Changes in proto-oncogene activity in the testis of the frog, Rana esculenta, during the annual reproductive cycle.

Proto-oncogenes are said to influence the regulation of cellular growth and differentiation. Myc, Fos, Jun, and Mos protein localization has been studied by immunocytochemistry in the testis of the frog, Rana esculenta, during the annual reproductive cycle. Oncoproteins have been localized in the primary and secondary (I and II) spermatogonia (SPG). Myc and Mos also appear in I and II spermatocytes (SPC) while Jun appears in II SPC. Myc, Fos, and Jun in SPG translocate in the nucleus during the periods of active spermatogenesis. Myc, Fos, and Jun are also localized in Sertoli cells. Fos is present in interstitial cells during the period characterized by the androgen peak which precedes the sharp increase of estradiol. It is suggested that proto-oncogene activity exerts a regulatory role in steroidogenesis and spermatogenesis.

Androgens↗
PubMed1995

Chicken GnRH-II and salmon GnRH effects on plasma and testicular androgen concentrations in the male frog, Rana esculenta, during the annual reproductive cycle.

In the frog, Rana esculenta, two molecular forms of GnRH, coeluting with chicken (c) GnRH-II and salmon (s) GnRH, have been detected using HPLC and radioimmunoassay. Mammalian (m) GnRH seems to be also present. In amphibians the role of cGnRH-II seems to be primarily the involvement in the regulation of neuroendocrine processes and, while the mGnRH has been postulated to act as a neurotransmitter and/or neuromodulator, the activity of sGnRH-like material has not been investigated. Therefore, we have treated the frogs with single or multiple injections of cGnRH-II or sGnRH (6 micrograms) or both peptides (6 micrograms of each) to detect differences in the response measured as testicular or plasma androgen (testosterone plus 5 alpha-dihydrotestosterone) concentration during the annual reproductive cycle. The basal profile of testicular and plasma androgen shows that the spring peak disappeared in control animals given multiple injections and kept in short-term captivity. We show in the treatment with cGnRH-II and/or sGnRH that the effects of the peptides depend on the season, the experimental design, and the tissue in which androgen levels were measured. In particular, both peptides strongly stimulate androgen production during the autumn-winter period, the time of the greater response to the GnRHs when basal levels of steroids are highest.

Analysis of Variance↗
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

Seasonal fluctuations of androgen-binding activity in the testis of the frog, Rana esculenta.

An androgen-binding activity has been identified in nuclear extracts of the testis of the frog, Rana esculenta. A single class of high affinity (Kd = 2.5 +/- 0.6 x 10(-9) M), low-capacity binding sites was found. The binding was specific for androgens; 17 beta-estradiol displaced [3H]testosterone with an ID50 of 0.1 microM. Cytosolic binding activity has a low affinity and a high capacity and lacks specificity. The seasonal fluctuations in binding capacity did not correlate with the androgen peaks in plasma and testes between February and June and in September; periods coinciding with the resumption of spermatogenesis and the development of spermatids, respectively. The present data strongly support androgenic control of intratesticular function in vertebrates generally.

Androgens↗
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↗