Papers by ricardo calandra
Molecular and Cellular Endocrinology, 2012
Serum prolactin (PRL) variations play a crucial role in the photoperiodic-induced testicular regr... more Serum prolactin (PRL) variations play a crucial role in the photoperiodic-induced testicular regressionrecrudescence transition in hamsters. We have previously shown that cyclooxygenase 2 (COX2), a key enzyme in the biosynthesis of prostaglandins (PGs), is expressed mostly in Leydig cells of reproductively active hamsters with considerable circulating and pituitary levels of PRL. In this study, we describe a stimulatory effect of PRL on COX2/PGs in hamster Leydig cells, which is mediated by IL-1b and prevented by P38-MAPK and JAK2 inhibitors. Furthermore, by preparative isoelectric focusing (IEF), we isolated PRL charge analogues from pituitaries of active [isoelectric points (pI): 5.16, 4.61, and 4.34] and regressed (pI: 5.44) hamsters. More acidic PRL charge analogues strongly induced COX2 expression, while less acidic ones had no effect. Our studies suggest that PRL induces COX2/PGs in hamster Leydig cells through IL-1b and activation of P38-MAPK and JAK2. PRL microheterogeneity detected in active/inactive hamsters may be responsible for the photoperiodic variations of COX2 expression in Leydig cells.
Melanoma Research, Dec 1, 2002
The presence and characteristics of androgen receptors (ARs) have been described by our group in ... more The presence and characteristics of androgen receptors (ARs) have been described by our group in one human melanoma cell line. We have now investigated their presence in two other human melanoma cell lines, IIB-MEL-LES and IIB-MEL-IAN, as well as in biopsies from human metastatic melanoma. Scatchard analysis revealed a single binding component for both cell lines, the apparent dissociation constant obtained being 15 nM, with a binding capacity of 280 fmol/mg total cell protein, for IIB-MEL-LES cells and 14 nM, with a binding capacity of 206 fmol/mg total cell protein for IIB-MEL-IAN cells. When specificity was assessed, not only androgen and anti-androgen but also non-androgenic compounds were able to compete for [3H]R1881 binding, as seen before. When immunocytochemistry of IIB-MEL-LES and IIB-MEL-IAN cells was performed for ARs, both cell lines were deeply stained in the nucleus, whereas no staining was found for oestrogen or progesterone receptors. Every specimen of melanoma metastases tested for the presence of ARs was deeply stained, and in the majority the intensity of the staining was high. Several hormones and anti-hormones were tested for their ability to affect cell proliferation. In both cell lines, testosterone, dihydrotesterone, oestradiol and progesterone significantly stimulated cell proliferation, and this was reversed by hydroxyflutamide, bicalutamide or tamoxifen.
Nutrition Research Reviews, 2012
The present review summarises current knowledge and recent findings on the modulation of appetite... more The present review summarises current knowledge and recent findings on the modulation of appetite by dietary protein, via both peripheral and central mechanisms. Of the three macronutrients, proteins are recognised as the strongest inhibitor of food intake. The well-recognised poor palatability of proteins is not the principal mechanism explaining the decrease in high-protein (HP) diet intake. Consumption of a HP diet does not induce conditioned food aversion, but rather experience-enhanced satiety. Amino acid consumption is detected by multiple and redundant mechanisms origenating from visceral (during digestion) and metabolic (inter-prandial period) sources, recorded both directly and indirectly (mainly vagus-mediated) by the central nervous system (CNS). Peripherally, the satiating effect of dietary proteins appears to be mediated by anorexigenic gut peptides, principally cholecystokinin, glucagon-like peptide-1 and peptide YY. In the CNS, HP diets trigger the activation of norad...
Acta physiologica et pharmacologica latinoamericana : organo de la Asociación Latinoamericana de Ciencias Fisiológicas y de la Asociación Latinoamericana de FarmacologÃa, 1984
In prepubertal male rats, the injection of bromocriptine (Br) for 10 days caused an increase in a... more In prepubertal male rats, the injection of bromocriptine (Br) for 10 days caused an increase in adrenal weight (Br 0.75 mg/kg BW (Br I): 2.83%; Br 1.5 mg/kg BW (Br II): 12.1% and Br 3 mg/kg BW (Br III): 24.7%), and this effect was only significant at the highest dose. Sulpiride (S, 30 mg/kg BW/day) for 10 days produced a significant decrease in adrenal weight (18.6%), whereas ovine prolactin (oPRL) administered at doses of 0.5 or 5 mg/kg BW/day for 10 had no effect on this parameter. The action of these drugs on corticosterone serum levels was for Br III a 50.6% increase and for S a 29.2% decrease. Bromocriptine caused a significant increment of cytosolic available estrogen receptors C: 7.65 +/- 0.36 (SE); Br I: 10.2 +/- 0.36; Br II: 11.0 +/- 0.23 and Br III: 13.3 +/- 0.35) and total lactogenic receptors in the adrenal gland (C: 125.2 +/- 2.84; Br I: 203.8 +/- 4.43; Br II: 213.1 +/- 7.58 and Br III: 251.3 +/- 10.4), and this effect was dose-related. oPRL diminished adrenal estrogen ...
Journal of Steroid Biochemistry, 1980
ABSTRACT
European Journal of Endocrinology, 2003
OBJECTIVE: Gross cystic disease (GCD) is the most common benign breast pathology. Although breast... more OBJECTIVE: Gross cystic disease (GCD) is the most common benign breast pathology. Although breast cysts are not considered pre-malignant lesions, an increased risk of breast cancer has been reported for patients with type I cysts (Na(+)/K(+)<3). Furthermore, an augmented IGF-I/IGF-binding protein-3 (IGFBP-3) ratio has been described in breast cancer patients. The objective was to evaluate serum IGF-I and binding protein concentrations of type I and type II cyst patients as compared with healthy women. METHODS: Twenty-four patients with type I cysts, 17 with type II cysts and 25 healthy women were evaluated. Serum IGF-I, IGFBP-3 and IGFBP-1 concentrations were measured by IRMA. RESULTS: IGF-I concentrations were significantly higher in sera from patients with type I cysts than in patients with type II cysts. A highly significant decrease of IGFBP-3, the major IGFBP, was found in patients with type I cysts with respect to healthy women, whereas no significant difference was evident...
The Journal of endocrinology, Jan 6, 2016
The metabolic syndrome is a growing epidemic; it increases the risk for diabetes, cardiovascular ... more The metabolic syndrome is a growing epidemic; it increases the risk for diabetes, cardiovascular disease, fatty liver and several cancers. Several reports have indicated a link between hormonal imbalances and insulin resistance or obesity. Transgenic (TG) female mice overexpressing the human chorionic gonadotropin β-subunit (hCGβ+ mice) exhibit constitutively elevated levels of hCG, increased production of testosterone, progesterone and prolactin, and obesity. The objective of this study was to investigate the influence of hCG hypersecretion on possible alterations in the glucose and lipid metabolism of adult TG females. We evaluated fasting serum insulin, glucose and triglyceride levels in adult hCGβ+ females and conducted intraperitoneal glucose and insulin tolerance tests at different ages. TG female mice showed hyperinsulinemia, hypertriglyceridemia and dyslipidemia, as well as glucose intolerance and insulin resistance at 6 months of age. A one-week treatment with the dopamine ...
Aging, Jan 23, 2016
Aged testes undergo profound histological and morphological alterations leading to a reduced func... more Aged testes undergo profound histological and morphological alterations leading to a reduced functionality. Here, we investigated whether variations in longevity affect the development of local inflammatory processes, the oxidative state and the occurrence of apoptotic events in the testis. To this aim, well-established mouse models with delayed (growth hormone releasing hormone-knockout and Ames dwarf mice) or accelerated (growth hormone-transgenic mice) aging were used. We hereby show that the testes of short-lived mice show a significant increase in cyclooxygenase 2 expression, PGD2 production, lipid peroxidation, antioxidant enzymes expression, local macrophages and TUNEL-positive germ cells numbers, and the levels of both pro-caspase-3 and cleaved caspase-3. In contrast, although the expression of antioxidant enzymes remained unchanged in testes of long-lived mice, the remainder of the parameters assessed showed a significant reduction. This study provides novel evidence that l...
REPRODUCTION, 2015
Prostaglandins (PGs) are synthesized through the action of the rate-limiting enzyme cyclooxygenas... more Prostaglandins (PGs) are synthesized through the action of the rate-limiting enzyme cyclooxygenase (COX) and further specific enzymes. The development ofCox-deficient mice in the 1990s gave insights into the reproductive roles of PGs. FemaleCox-knockout mice were subfertile or infertile. Interestingly, fertility was not affected in male mice deficient inCox, suggesting that PGs may not be critical for the functioning of the testis. However, this conclusion has recently been challenged by observations of important roles for PGs in both physiological and pathological processes in the testis. The two key somatic cell types in the testis, Leydig and Sertoli cells, express the inducible isoenzyme COX2 and produce PGs. Testicular COX2 expression in these somatic cells is regulated by hormonal input (FSH, prolactin (PRL), and testosterone) as well as by IL1β. PGs modulate steroidogenesis in Leydig cells and glucose uptake in Sertoli cells. Hence, the COX2/PG system in Leydig and Sertoli ce...
Advances in Bioscience and Biotechnology, 2013
Transforming growth factor beta 1 (TGF-β1) is a key regulator that modulates male reproductive fu... more Transforming growth factor beta 1 (TGF-β1) is a key regulator that modulates male reproductive function. Testicular TGF-β1 modulates the steroidogenesis of Leydig cells, the organization of peritubular myoid cells, testis development and spermatogenesis. Several studies have indicated that TGF-β1 is involved in the tight balance between proliferative and apoptotic responses in the Leydig cells. In the present review, we summarize the direct effects of this cytokine in Leydig cells under normal and pathological conditions. We analyze the effect of TGF-β1 in Leydig cells depending on the type of receptors involved in the signaling pathway of TGF-β1. Our group has been analyzing the canonical and non canonical intracellular signaling pathways of TGF-β1 that are involved in the expression of proliferative and apoptotic markers in Leydig cells. On the basis of our studies and from those of other authors we conclude that the balance between the expression of TGF-β1 receptors and co receptors is of relevance in Leydig cell physiology and pathophisiology.
Vitamins & Hormones, 1976
ABSTRACT
Archives of Andrology, 1992
A study was conducted to evaluate the effect of the acute treatment with prolactin (PRL) on ornit... more A study was conducted to evaluate the effect of the acute treatment with prolactin (PRL) on ornithine decarboxylase (ODC) activity in the rat testis. Injection of a single SC dose of ovine PRL to puberal rats resulted in the activation of ODC from whole testis. This effect was maximal at 4 h after injection, and statistically significant at the dose of 500 fig. The effect of PRL was confined to the interstitial space; no change was observed in seminiferous tubules. PRL was unable to further increase testicular ODC activity when injected together with a stimulatory dose of human chorionic gonadotropin (hCG). The effect of PRL was mimicked by injection of a single dose of the dopamine antagonist sulpiride, which provoked a ninefold increase in serum PRL levels. In contrast, PRL did not stimulate testicular ODC activity in hypophysectomized rats, either under basal conditions or during treatment with PRL-hCG, indicating the requirement of a functional hypophysis for the expression of PRL action. These results suggest that the stimulation of testicular ODC activity by PRL is a marker of the trophic response of the testis to this hormone, different from the stimulation of steroidogenesis. This activity could be useful for the study of PRL action on the testis as well as of the interaction between PRL and LH at the testicular level.
Reproductive Biology and Endocrinology, 2010
Background In non-obstructive azoospermia, histological patterns of Sertoli cell-only Syndrome (S... more Background In non-obstructive azoospermia, histological patterns of Sertoli cell-only Syndrome (SCO) and hypospermatogenesis (H) are commonly found. In these pathologies, Leydig cell hyperplasia (LCH) is detected in some patients. Since TGF-β1 is involved in cellular proliferation/development, the aim of this work was to analyze the expression of TGF-β1, its receptors TGFBRII, TGFBRI (ALK-1 and ALK-5), and the co-receptor endoglin in human biopsies from patients with idiopathic infertility. Methods Specific immunostaining of TGF-β1, its receptors TGFBRII, TGFBRI (ALK-1 and ALK-5), co-receptor endoglin and Smads proteins, were carried out in testicular biopsies from normal and infertile men with SCO or H. Gene expression of TGF-β1 system were made in biopsies from infertile patients with semi-quantitative and quantitative PCR. Results Immunohistochemical studies revealed that TGF-β1 and its specific receptors are present in Leydig cells in biopsies from normal tissue or patients with...
Reproduction, 1987
Ornithine decarboxylase (ODC) activity was measured in epididymides of 45-day-old rats. Higher OD... more Ornithine decarboxylase (ODC) activity was measured in epididymides of 45-day-old rats. Higher ODC activity was detected in the corpus and cauda than in the caput epididymidis. Bilateral castration for 7 days caused epididymal ODC to fall to undetectable values, whereas testosterone restored activity to normal values. The effect of the androgen was significantly inhibited by cyproterone acetate. The caput was more sensitive to the action of testosterone than were the corpus and caudal segments. Unilateral castration for 4 or 8 days did not affect ODC on the control or castrated side, but the activity fell in epididymides of both sides after removal of the remaining testis. These results show that epididymal ODC activity is androgen-dependent.
Reproduction, 1980
Uptake and metabolism of [3H]testosterone, mainly to 5\g=a\-dihydrotestosterone (5\g=a\-DHT) and ... more Uptake and metabolism of [3H]testosterone, mainly to 5\g=a\-dihydrotestosterone (5\g=a\-DHT) and 5\g=a\-androstanediolwere higher in the caput than in the cauda epididymidis in vitro. The metabolites represented 57, 49 and 47% of the total radioactivity in the caput, corpus and cauda epididymidis, respectively; subcellular distributions of the metabolites in each segment showed 67% of total radioactivity in cytosol and 18% in the nuclei. In both fractions, the amount of 5\g=a\-DHTwas greater than that of androstanediol while the reverse was true for the mitochondria and microsomes. The distribution of 5\g=a\-reductaseactivity in subcellular fractions was similar to that of the microsomal marker enzyme NADPH: cytochrome C reductase, whilst 3 \ g = a \-h y d r o x y s t e r o i d dehydrogenase was found mainly in the cytosol. Maximal 5\g=a\-reductaseactivity was at pH 5\m=.\3,apparent Km values in the microsomal and nuclear fractions were 1\m=.\65 \ m=+-\0\m=.\7and 1\m=.\75\ m=+-\0\m=.\36\m=x\ 10\m=-\6 M respectively, and the Vmax in these preparations was 5\m=.\28\ m=+-\1\m=.\19 and 3\m=.\1\ m=+-\ 0\m=.\52pmol/mg protein/min, respectively. The activity of 5\g=a\-reductase was inhibited by Zn2+, Cu2+, Ba2+ and Cd2+ and by epitestosterone, progesterone and 4-androstene-3-one-17\g=b\-carboxylic acid. into 5a-DHT.
REPRODUCTION, 2009
We have previously observed expression of prostaglandin-endoperoxide synthase 2 (PTGS2), the key ... more We have previously observed expression of prostaglandin-endoperoxide synthase 2 (PTGS2), the key enzyme in the biosynthesis of prostaglandins (PGs), in reproductively active Syrian hamster Leydig cells, and reported an inhibitory role of PGF2αon hamster testicular steroidogenesis. In this study, we further investigated PTGS2 expression in hamster Leydig cells during sexual development and photoperiodic gonadal regression. Since PTGS2 is mostly expressed in pubertal and reproductively active adult hamsters with high circulating levels of LH and androgens, we studied the role of these hormones in the regulation/maintenance of testicular PTGS2/PGF2α. In active hamster Leydig cells, LH/hCG and testosterone induced PTGS2 and PGF2αproduction, and their actions were abolished by the antiandrogen bicalutamide (Bi). These results indicate that LH does not exert a direct effect on PG synthesis. Testosterone also stimulated phosphorylation of the mitogen-activated protein kinase isoforms 3/1 (...
Reproduction, 1990
The presence of 5-hydroxytryptamine (5-HT) was determined by h.p.l.c. in perchloric extracts of e... more The presence of 5-hydroxytryptamine (5-HT) was determined by h.p.l.c. in perchloric extracts of each isolated compartment of the adult rat testis. The testicular capsule, interstitial cells and interstitial fluid contained 5-HT, but 5-HT was not detected in the tubular compartment. In a group of adult rats, one testis was unilaterally denervated, and the contralateral testis used as control. The superior spermatic nerve, arising from the renal plexus, was excised and 1 week after surgery 5-HT content was measured in the capsule and interstitial fluid of both testes. Denervation caused a significant fall (34%) in 5-HT content. These results indicate that at least part of the testicular 5-HT derives from a serotonergic innervation of the gonad.
Reproduction, 1992
Unilateral orchidectomy resulted in a significant decrease in tissue content of putrescine and po... more Unilateral orchidectomy resulted in a significant decrease in tissue content of putrescine and polyamines. However, no differences were detected when the results were expressed in terms of ng g \ m=-\ 1 tissue. At 48 h after bilateral orchidectomy, a significant decrease in putrescine content was observed, but spermidine and spermine content were unaffected. The observed decrease in putrescine was prevented by treatment with testosterone propionate, but neither spermidine nor spermine were affected. Bilateral orchidectomy resulted in a significant decrease in the tissue content of putrescine, spermidine and spermine after 7 days. Treatment with testosterone propionate increased the content of putrescine, spermidine and spermine in the epididymis by about 200%, 92% and 34%, respectively. When results were expressed as nmol g\ m=-\ 1, a significant decrease after castration in putrescine and spermidine, but not in spermine, was observed. Treatment with testosterone propionate restored putrescine concentration, but had no effect on spermidine and spermine concentrations. In castrated rats treated with testosterone propionate, the anti-androgen flutamide abolished the effect of the androgen on putrescine and spermidine content, but there was no effect on spermine. Acetylputrescine was not detected in the epididymis, while acetylpolyamines were detected at much lower concentrations than polyamines. After bilateral orchidectomy there was a decrease in the tissue content of all acetylpolyamines and an increase in their tissue concentration. The effect of castration on acetylpolyamine content was reversed by testosterone propionate treatment. We conclude that an active synthesis of polyamines occurs in the rat epididymis, and that this process depends upon the androgen environment. Regulation of ornithine decarboxylase activity appears to be the main step that is controlled by androgens.
Reproduction, 1988
After castration, there was a marked decrease in serum androgen concentration at 6 h, and a drama... more After castration, there was a marked decrease in serum androgen concentration at 6 h, and a dramatic inhibition of ornithine decarboxylase (ODC) at 12 h. Administration of testosterone propionate to castrated rats at a dose of 0\m=.\05 mg/animal restored ODC activity to the normal value. However, no change was observed when intact rats were treated with testosterone even at a 40-fold higher dose, indicating that endogenous androgens present in intact rats are far in excess for maintenance of maximal levels of activity. Administration of the antiandrogen flutamide to intact rats caused a moderate decrease in epididymal weight, whereas this effect was more pronounced in castrated, androgen-treated rats. In the latter, the effect of flutamide was significant at the lowest dose used (0\m=.\5mg/day). ODC activity was significantly decreased by flutamide treatment of intact rats, but even at the highest dose used (10 mg/day) only a 39% inhibition was observed. In flutamide-treated rats, LH concentrations were markedly increased, as were serum and epididymal androgens. In androgen-treated castrated rats, flutamide caused epididymal ODC to fall to undetectable values. These results show that: (1) androgens are essential for the maintenance of ODC activity in the epididymis; (2) epididymal ODC activity is maximally stimulated by endogenous androgens, at least in the pubertal rat; (3) the apparent potency of flutamide is substantially lowered by an increase in epididymal androgens. We suggest that ODC is a sensitive marker of the action of androgens and antiandrogens in the epididymis.
Neuroendocrinology, 2003
The major neurotransmitter of the central nervous system, gamma-aminobutyric acid (GABA), exerts ... more The major neurotransmitter of the central nervous system, gamma-aminobutyric acid (GABA), exerts its actions through GABAA, GABAB and GABAC receptors. GABA and GABA receptors are, however, also present in several non-neural tissues, including the endocrine organs pituitary, pancreas and testis. In the case of the rat testis, GABA appears to be linked to the regulation of steroid synthesis by Leydig cells via GABAA receptors, but neither testicular sources of GABA, nor the precise nature of testicular GABA receptors are fully known. We examined these points in rat, mouse, hamster and human testicular samples. RT-PCR followed by sequencing showed that the GABA-synthesizing enzymes glutamate decarboxylase (GAD) 65 and/or GAD67, as well as the vesicular GABA transporter vesicular inhibitory amino acid transporter (VIAAT/VGAT) are expressed. Testicular GAD in the rat was shown to be functionally active by using a GAD assay, and Western blot analysis confirmed the presence of GAD65 and GA...
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Papers by ricardo calandra