Biomedical subjects

Angel Nadal

Publications and source records attributed to Angel Nadal.

17 recordsLinked to original sources

PubMed2006

Rapid endocrine disruption: environmental estrogen actions triggered outside the nucleus.

An exogenous substance is defined as an endocrine disrupter chemical (EDC) if it alters the function of the endocrine system provoking adverse health effects. Environmental estrogens are the most studied EDCs. They follow the same mechanisms of action as the gonadal hormone 17beta-estradiol. Up to now, the estrogenicity of environmental estrogenic pollutants has been based on the property of these compounds to bind to estrogen receptors (ERs), either ERalpha or ERbeta, and to act subsequently as transcription factors when binding to the estrogen response element (ERE) in the DNA. All the estrogenic bioassays currently used are based on this mechanism of action. New evidence indicates that the definition of estrogenicity for a chemical should take into account other estrogen receptors as well as new signaling pathways. These include the activation of additional transcription factors as well as the action of xenoestrogens through estrogen receptors located outside the nucleus: in the plasma membrane, mitochondria and probably the cytosol. Therefore, new estrogenic bioassays should be developed to include the novel concept of rapid endocrine disruption.

Animals↗

PubMed2006

Activation of cannabinoid CB1 receptors induces glucose intolerance in rats.

Recent reports have described the presence of cannabinoid CB1 receptors in pancreatic islets. Here we show that administration of the endogenous cannabinoid anandamide or the selective cannabinoid CB1 receptor agonist Arachidonyl-2'-chloroethylamide (ACEA) results in glucose intolerance after a glucose load. This effect is reversed by the selective cannabinoid CB1 receptor antagonist N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251). These results suggest that targeting cannabinoid CB1 receptors may serve as new therapeutic alternatives for metabolic disorders such as diabetes.

Animals↗

PubMed2006

Cannabinoid receptors regulate Ca(2+) signals and insulin secretion in pancreatic beta-cell.

Insulin is the main hormone involved in the regulation of glycaemia, its impaired secretion is a hallmark of type I and type II diabetic individuals. Additionally, insulin is involved in lipogenesis and weight gain, provoking an anorexigenic action. The endocannabinoid system contributes to the physiological regulation of energy balance, food intake and lipid and glucose metabolisms. Despite that, an experimental link between the endocannabinoid system and the endocrine pancreas has not yet been described. Using quantitative real-time PCR and immunocytochemistry, we have demonstrated the existence of both CB1 and CB2 receptors in the endocrine pancreas. While the CB1 receptor is mainly expressed in non-beta-cells, the CB2 type exists in beta- and non-beta-cells within the islet. The endocannabinoid 2-arachidonylglycerol (2-AG) through CB2 receptors regulates [Ca(2+)](i) signals in beta-cells and as a consequence, it decreases insulin secretion. This effect may be a new component involved in the orexigenic effect of endocannabinoids and constitutes a potential target for pharmacologic manipulation of the energy balance.

Animals↗

PubMed2006

The estrogenic effect of bisphenol A disrupts pancreatic beta-cell function in vivo and induces insulin resistance.

The function of the pancreatic beta-cell is the storage and release of insulin, the main hormone involved in blood glucose homeostasis. The results in this article show that the widespread environmental contaminant bisphenol-A (BPA) imitates 17beta-estradiol (E2) effects in vivo on blood glucose homeostasis through genomic and nongenomic pathways. The exposure of adult mice to a single low dose (10 microg/kg) of either E2 or BPA induces a rapid decrease in glycemia that correlates with a rise of plasma insulin. Longer exposures to E2 and BPA induce an increase in pancreatic beta-cell insulin content in an estrogen-receptor-dependent manner. This effect is visible after 2 days of treatment and starting at doses as low as 10 microg/kg/day. After 4 days of treatment with either E2 or BPA, these mice developed chronic hyperinsulinemia, and their glucose and insulin tolerance tests were altered. These experiments unveil the link between environmental estrogens and insulin resistance. Therefore, either abnormal levels of endogenous estrogens or environmental estrogen exposure enhances the risk of developing type 2 diabetes mellitus, hypertension, and dyslipidemia.

Animals↗

PubMed2006

Glucose induces opposite intracellular Ca2+ concentration oscillatory patterns in identified alpha- and beta-cells within intact human islets of Langerhans.

Homeostasis of blood glucose is mainly regulated by the coordinated secretion of glucagon and insulin from alpha- and beta-cells within the islets of Langerhans. The release of both hormones is Ca(2+) dependent. In the current study, we used confocal microscopy and immunocytochemistry to unequivocally characterize the glucose-induced Ca(2+) signals in alpha- and beta-cells within intact human islets. Extracellular glucose stimulation induced an opposite response in these two cell types. Although the intracellular Ca(2+) concentration ([Ca(2+)](i)) in beta-cells remained stable at low glucose concentrations, alpha-cells exhibited an oscillatory [Ca(2+)](i) response. Conversely, the elevation of extracellular glucose elicited an oscillatory [Ca(2+)](i) pattern in beta-cells but inhibited low-glucose-induced [Ca(2+)](i) signals in alpha-cells. These Ca(2+) signals were synchronic among beta-cells grouped in clusters within the islet, although they were not coordinated among the whole beta-cell population. The response of alpha-cells was totally asynchronic. Therefore, both the alpha- and beta-cell populations within human islets did not work as a syncitium in response to glucose. A deeper knowledge of alpha- and beta-cell behavior within intact human islets is important to better understand the physiology of the human endocrine pancreas and may be useful to select high-quality islets for transplantation.

Adolescent↗

PubMed2005

Bioluminescence imaging of nuclear calcium oscillations in intact pancreatic islets of Langerhans from the mouse.

The stimulus-secretion coupling for insulin secretion by pancreatic beta cells in response to high glucose involves synchronic cytosolic calcium oscillations driven by bursting electrical activity. Calcium inside organelles can regulate additional functions, but analysis of subcellular calcium signals, specially at the single cell level, has been hampered for technical constrains. Here we have monitored nuclear calcium oscillations by bioluminescence imaging of targeted aequorin in individual cells within intact islets of Langerhans as well as in the whole islet. We find that glucose generates a pattern of nuclear calcium oscillations resembling those found in the cytosol. Some cells showed synchronous nuclear calcium oscillations suggesting that the islet of Langerhans may also regulate the activation of Ca(2+)-responsive nuclear processes, such as gene transcription, in a coordinated, synchronic manner. The nuclear Ca(2+) oscillations are due to bursting electrical activity and activation of plasma membrane voltage-gated Ca(2+) channels with little or no contribution of calcium release from the intracellular Ca(2+) stores. Irregularities in consumption of aequorins suggests that depolarization may generate formation of steep Ca(2+) gradients in both the cytosol and the nucleus, but further research is required to investigate the role of such high [Ca(2+)] microdomains.

Aequorin↗

PubMed2005

Low doses of bisphenol A and diethylstilbestrol impair Ca2+ signals in pancreatic alpha-cells through a nonclassical membrane estrogen receptor within intact islets of Langerhans.

Glucagon, secreted from pancreatic alpha-cells integrated within the islets of Langerhans, is involved in the regulation of glucose metabolism by enhancing the synthesis and mobilization of glucose in the liver. In addition, it has other extrahepatic effects ranging from lipolysis in adipose tissue to the control of satiety in the central nervous system. In this article, we show that the endocrine disruptors bisphenol A (BPA) and diethylstilbestrol (DES), at a concentration of 10(-9) M, suppressed low-glucose-induced intracellular calcium ion ([Ca2+]i) oscillations in alpha-cells, the signal that triggers glucagon secretion. This action has a rapid onset, and it is reproduced by the impermeable molecule estradiol (E2) conjugated to horseradish peroxidase (E-HRP). Competition studies using E-HRP binding in immunocytochemically identified alpha-cells indicate that 17beta-E2, BPA, and DES share a common membrane-binding site whose pharmacologic profile differs from the classical ER. The effects triggered by BPA, DES, and E2 are blocked by the G alpha i- and G alpha o-protein inhibitor pertussis toxin, by the guanylate cyclase-specific inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, and by the nitric oxide synthase inhibitor N-nitro-L-arginine methyl ester. The effects are reproduced by 8-bromo-guanosine 3',5'-cyclic monophosphate and suppressed in the presence of the cGMP-dependent protein kinase inhibitor KT-5823. The action of E2, BPA, and DES in pancreatic alpha-cells may explain some of the effects elicited by endocrine disruptors in the metabolism of glucose and lipid.

Animals↗

PubMed2004

Disentangling the molecular mechanisms of action of endogenous and environmental estrogens.

The gonadal hormone 17beta-estradiol is involved in numerous cellular processes. In many cases, 17beta-estradiol actions are imitated by synthetic and natural chemicals in the environment. Their actions differ depending on the target tissue, the receptors involved and the molecular pathways activated. The plethora of estrogenic actions is triggered by different receptors and other specific structures that activate different signalling pathways. This amount of information may lead to a maze of effects triggered by endogenous and environmental estrogens that we intend to clarify in this review. Understanding the variety of estrogen receptors, their different locations and the signalling pathways activated by estrogenic ligands is fundamental for understanding the diversity of actions that estrogens have in different tissues and cells.

Animals↗

PubMed2004

Oestradiol rapidly inhibits Ca2+ signals in ciliary neurons through classical oestrogen receptors in cytoplasm.

Oestrogen plays a key role in a great variety of actions in the nervous system, either through classical or alternative pathways. The classical pathways are initiated after oestrogen binding to the oestrogen receptors ERalpha or ERbeta, which translocate from the cytoplasm to the nucleus and act there as transcription factors. Alternative pathways are initiated at the plasma membrane and cytoplasm, via binding to classical or non-classical ERs. Using isolated ciliary ganglion neurons from the chick embryo and Ca2+ imaging, we demonstrated that a 10-min exposure to 17beta-oestradiol reduces Ca2+ influx through the plasma membrane. This effect was not reproduced by oestradiol conjugated to bovine serum albumin, which does not cross the plasma membrane, indicating that 17beta-oestradiol was acting intracellularly. ERalpha was detected in the cytoplasm by immunostaining and its involvement in the regulation of Ca2+ influx by ICI182,780 inhibition. The phosphatidylinositol-3 kinase (Pi3-kinase) inhibitor wortmannin and the nitric oxide synthase (NOS) inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) both blocked the oestradiol effect. The oestradiol effect was reproduced by 8Br-cGMP and abolished in the presence of the cGMP-dependent protein kinase (PKG) inhibitor KT5823. Our study indicates that 17beta-oestradiol can regulate Ca2+ influx via PI3-kinase, NOS and PKG after activation of cytoplasmic ER.

Animals↗

PubMed2004

Estrogen and xenoestrogen actions on endocrine pancreas: from ion channel modulation to activation of nuclear function.

17beta-Estradiol elicits a rapid opposite effect on [Ca2+]i in alpha- and beta-cells within intact islets of Langerhans. In beta-cells, physiological concentrations of the gonadal hormone decreases KATP channel activity in synergy with glucose, leading to a membrane depolarization that opens voltage-gated Ca2+ channels, potentiating Ca2+ signals. As a consequence insulin release is enhanced and transcription factor CREB is activated in a Ca(2+)-dependent manner. In glucagon-containing alpha-cells, 17beta-estradiol provokes the abolishment of Ca2+ oscillations generated by low glucose, a situation that should decrease glucagon release. In both types of cells the second messenger involved is cGMP. The estrogen receptor involved is located in the plasma membrane and has a pharmacological profile unrelated to classical estrogen receptors ERalpha and ERbeta. For that reason, it has been named non-classical membrane estrogen receptor (ncmER). Although the physiological roles of this receptor are still unknown, it may be implicated in the responses of the endocrine pancreas to the physiological and pathological changes of 17beta-estradiol.

Animals↗

PubMed2004

Novel players in pancreatic islet signaling: from membrane receptors to nuclear channels.

Glucose and other nutrients regulate many aspects of pancreatic islet physiology. This includes not only insulin release, but also insulin synthesis and storage and other aspects of beta-cell biology, including cell proliferation, apoptosis, differentiation, and gene expression. This implies that in addition to the well-described signals for insulin release, other intracellular signaling mechanisms are needed. Here we describe the role of global and local Ca(2+) signals in insulin release, the regulation of these signals by new membrane receptors, and the generation of nuclear Ca(2+) signals involved in gene expression. An integrated view of these pathways should improve the present description of the beta-cell biology and provide new targets for novel drugs.

Animals↗

PubMed2003

Bovine subcommissural organ displays spontaneous and synchronous intracellular calcium oscillations.

The subcommissural organ (SCO) is an ependymal brain gland that secretes into the cerebrospinal fluid glycoproteins that polymerize, forming Reissner's fiber (RF). The SCO-RF complex seems to be involved in vertebrate nervous system development, although its role in adults is unknown. Furthermore, its physiology is still greatly undetermined, and little is known about the release control of SCO secretion and the underlying intracellular mechanisms. In this report, we show that up to 90% of 3-5-day-old in vitro SCO cells from both intact and partially-dispersed SCO explants displayed spontaneous cytosolic Ca2+ oscillations. The putative role of these spontaneous calcium oscillations in SCO secretory activity is discussed taking into consideration several previous findings. Two distinct subpopulations of SCO cells were detected, each one containing cells with synchronized calcium oscillations. A possible existence of different functional domains in SCO is therefore discussed. Oscillations persisted in the absence of extracellular Ca2+, indicating the major involvement of Ca2+ released from internal stores. Depolarization failed to induce intracellular calcium increases, although it disturbed the oscillation frequency, suggesting a putative modulator role of depolarizing agonists on the calcium oscillating pattern through voltage-gated calcium channels. Carbachol, a cholinergic agonist, evoked a switch in Ca2+ signaling from a calcium oscillating mode to a sustained and increased intracellular Ca2+ mode in 30% of measured cells, suggesting the involvement of acetylcholine in SCO activity, via a calcium-mediated response.

Analgesics, Non-Narcotic↗

PubMed2003

On-line analysis of gap junctions reveals more efficient electrical than dye coupling between islet cells.

Pancreatic beta-cells constitute a well-communicating multicellular network that permits a coordinated and synchronized signal transmission within the islet of Langerhans that is necessary for proper insulin release. Gap junctions are the molecular keys that mediate functional cellular connections, which are responsible for electrical and metabolic coupling in the majority of cell types. Although the role of gap junctions in beta-cell electrical coupling is well documented, metabolic communication is still a matter of discussion. Here, we have addressed this issue by use of a fluorescence recovery after photobleaching (FRAP) approach. This technique has been validated as a reliable and noninvasive approach to monitor functional gap junctions in real time. We show that control pancreatic islet cells did not exchange a gap junction-permeant molecule in either clustered cells or intact islets of Langerhans under conditions that allowed cell-to-cell exchange of current-carrying ions. Conversely, we have detected that the same probe was extensively transferred between islet cells of transgenic mice expressing connexin 32 (Cx32) that have enhanced junctional coupling properties. The results indicate that the electrical coupling of native islet cells is more extensive than dye communication. Dye-coupling domains in islet cells appear more restricted than previously inferred with other methods.

Animals↗

PubMed2003

Estradiol modulates acetylcholine-induced Ca2+ signals in LHRH-releasing GT1-7 cells through a membrane binding site.

Estrogen regulation of the female reproductive axis involves the rapid inhibition (< 30 min) of luteinizing hormone-releasing hormone (LHRH) secretion from hypothalamic neurons. This fast time-course suggests interactions with potential plasma membrane binding sites that could result in short-term effects on LHRH neurons. Because LHRH release is calcium dependent, we have studied the acute effects of 17beta-estradiol (E2) and estradiol-peroxidase (E-HRP) on the elevations of intracellular calcium ([Ca2+]i) induced by acetylcholine (ACh) in LHRH-producing GT1-7 cells. Exposure to ACh (1-100 micro m) induced transient increases of [Ca2+]i, whereas pretreatment with E2 or E-HRP (10 nm) for 2 min reduced this response by 50-60%. The effect was specific for E2 as neither 17alpha-estradiol (1 micro m) nor the synthetic antiestrogens ICI182 780 (1 micro m) or tamoxifen (1 micro m) elicited any change on the ACh-induced Ca2+ signal. Both the latency of the effect and the response to the membrane impermeant conjugate suggested a membrane-mediated mechanism. Such membrane binding sites for E2 in GT1-7 cells were demonstrated by visualizing the binding of E-HRP and estradiol-BSA-fluorescein isothiocyanate (E-BSA-FITC) conjugates. Competition studies showed that E-HRP binding was blocked by preincubation with E2, but not with 17alpha-E2, ICI182 780, tamoxifen or progesterone, indicating that the plasma membrane binding site is highly specific for E2 and exhibits a pharmacological profile different from classical estrogen receptors. We conclude that ACh-induced increase in [Ca2+]i in GT1-7 cells is modulated acutely by physiological E2 concentrations in a manner which is compatible with the existence of an estrogen-specific membrane binding site.

Acetylcholine↗

PubMed2002

Low doses of the endocrine disruptor bisphenol-A and the native hormone 17beta-estradiol rapidly activate transcription factor CREB.

Endocrine-disrupting chemicals (EDCs) are hormone-like agents present in the environment that alter the endocrine system of wildlife and humans. Most EDCs have potencies far below those of the natural hormone 17beta-E2 when acting through the classic estrogen receptors (ERs). Here, we show that the environmental estrogen Bisphenol-A and the native hormone 17beta-E2 activate the transcription factor, cAMP-responsive element binding protein (CREB) with the same potency. Phosphorylated CREB (P-CREB) was increased after only a 5-minute application of either BPA or 17beta-E2 in a calcium-dependent manner. The effect was reproduced by the membrane-impermeable molecule E2 conjugated to horseradish peroxidase (E-HRP). The increase in P-CREB was not modified by the anti-estrogen ICI 182,780. Therefore, low-dose of BPA activates the transcription factor CREB via an alternative mechanism, involving a non-classical membrane estrogen receptor. Because these effects are elicited at concentrations as low as 10(-9) M, this observation is of environmental and public health relevance.

Animals↗

PubMed2002

Nuclear KATP channels trigger nuclear Ca(2+) transients that modulate nuclear function.

Glucose, the principal regulator of endocrine pancreas, has several effects on pancreatic beta cells, including the regulation of insulin release, cell proliferation, apoptosis, differentiation, and gene expression. Although the sequence of events linking glycemia with insulin release is well described, the mechanism whereby glucose regulates nuclear function is still largely unknown. Here, we have shown that an ATP-sensitive K(+) channel (K(ATP)) with similar properties to that found on the plasma membrane is also present on the nuclear envelope of pancreatic beta cells. In isolated nuclei, blockade of the K(ATP) channel with tolbutamide or diadenosine polyphosphates triggers nuclear Ca(2+) transients and induces phosphorylation of the transcription factor cAMP response element binding protein. In whole cells, fluorescence in situ hybridization revealed that these Ca(2+) signals may trigger c-myc expression. These results demonstrate a functional K(ATP) channel in nuclei linking glucose metabolism, nuclear Ca(2+) signals, and nuclear function.

Adenosine Triphosphate↗

PubMed2002

A nonclassical estrogen membrane receptor triggers rapid differential actions in the endocrine pancreas.

Glucose homeostasis in blood is mainly maintained by insulin released from beta-cells and glucagon released from alpha-cells, both integrated within the pancreatic islet of Langerhans. The secretory processes in both types of cells are triggered by a rise in intracellular calcium concentration ([Ca2+](i)). In this study, rapid effects of the natural hormone E2 on [Ca2+](i) were studied in both types of cells within intact islets using laser scanning confocal microscopy. alpha- And beta-cells showed opposite [Ca2+](i) responses when stimulated with physiological concentrations of 17beta-E2. Although the estrogen produced an increase in the frequency of glucose-induced [Ca2+](i) oscillations in insulin-releasing beta-cells, it prevented the low glucose-induced [Ca2+](i) oscillations in glucagon-releasing alpha-cells. The effects of 17beta-E2 on alpha-cells were mimicked by the cGMP permeable analog 8bromo-cGMP and blocked by the cGMP-dependent protein kinase (PKG) inhibitor KT5823. Evidence indicated that these were membrane actions mediated by a nonclassical ER. Both effects were rapid in onset and were reproduced by 17beta-E2 linked to horseradish peroxidase, a cell-impermeable molecule. Furthermore, these actions were not blocked by the specific ER blocker ICI 182,780. Competition studies performed with 17beta-E2 linked to horseradish peroxidase binding in alpha-cells supported the idea that the membrane receptor involved is neither ERalpha nor ERbeta. Additionally, the binding site was shared by the neurotransmitters epinephrine, norepinephrine, and dopamine and had the same pharmacological profile as the receptor previously described for beta-cells. Therefore, rapid estrogen actions in islet cells are initiated by a nonclassical estrogen membrane receptor.

Alkaloids↗