Biomedical subjects

Bernat Soria

Publications and source records attributed to Bernat Soria.

11 recordsLinked to original sources

PubMed2004

Transforming growth factor (TGF)beta, fibroblast growth factor (FGF) and retinoid signalling pathways promote pancreatic exocrine gene expression in mouse embryonic stem cells.

Extracellular signalling cues play a major role in the activation of differentiation programmes. Mouse embryonic stem (ES) cells are pluripotent and can differentiate into a wide variety of specialized cells. Recently, protocols designed to induce endocrine pancreatic differentiation in vitro have been designed but little information is currently available concerning the potential of ES cells to differentiate into acinar pancreatic cells. By using conditioned media of cultured foetal pancreatic rudiments, we demonstrate that ES cells can respond in vitro to signalling pathways involved in exocrine development and differentiation. In particular, modulation of the hedgehog, transforming growth factor beta, retinoid, and fibroblast growth factor pathways in ES cell-derived embryoid bodies (EB) resulted in increased levels of transcripts encoding pancreatic transcription factors and cytodifferentiation markers, as demonstrated by RT-PCR. In EB undergoing spontaneous differentiation, expression of the majority of the acinar genes (i.e. amylase, carboxypeptidase A and elastase) was induced after the expression of endocrine genes, as occurs in vivo during development. These data indicate that ES cells can undergo exocrine pancreatic differentiation with a kinetic pattern of expression reminiscent of pancreas development in vivo and that ES cells can be coaxed to express an acinar phenotype by activation of signalling pathways known to play a role in pancreatic development and differentiation.

Activins↗

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↗

PubMed2004

Nutrients induce different Ca(2+) signals in cytosol and nucleus in pancreatic beta-cells.

Specific activation of Ca(2+)-dependent functions is achieved by the particular dynamics and local restriction of Ca(2+) signals. It has been shown that changes in amplitude, duration, or frequency of Ca(2+) signals modulate gene transcription. Thus, Ca(2+) variations should be finely controlled within the nucleus. Although a variety of mechanisms in the nuclear membrane have been demonstrated to regulate nuclear Ca(2+), the existence of an autonomous Ca(2+) homeostasis within the nucleus is still questioned. In the pancreatic beta-cell, besides their effect on insulin secretion, Ca(2+) messages generated by nutrients also exert their action on gene expression. However, the dynamics of these Ca(2+) signals in relation to nuclear function have been explored little in islet cells. In the current study, Ca(2+) changes both in the nucleoplasm and in the cytosol of INS-1 and pancreatic beta-cells were monitored using spot confocal microscopy. We show that nutrients trigger Ca(2+) signals of higher amplitude in the nucleus than in the cytosol. These amplitude-modulated Ca(2+) signals transmitted to the nucleus might play an important role in the control of gene expression in the pancreatic beta-cell.

ATP-Binding Cassette Transporters↗

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

Mitochondrial dysfunction is involved in apoptosis induced by serum withdrawal and fatty acids in the beta-cell line INS-1.

The potential toxic effects of high extracellular concentrations of fatty acids were tested in beta(INS-1)-cells cultured in the absence of serum, a condition known to alter cell survival in various systems. This may in part mimic the situation in type 1 or 2 diabetes where beta-cells are already insulted by various stressful conditions, such as cytokines and oxidative stress. Serum removal caused, over a 36-h period, oxidative stress and an early impairment of mitochondrial function, as revealed by increased superoxide production and markedly reduced mitochondrial membrane potential, but a lack of cytochrome c and apoptosis-inducing factor release in the cytosol. The fatty acids palmitate and oleate considerably accelerated the apoptosis process in serum-starved cells, as revealed by fluorescence-activated cell sorting analysis, morphological changes, chromatin condensation, DNA laddering, poly(ADP-ribose) polymerase cleavage, cytochrome c and apoptosis-inducing factor release, and increased levels of Bax and cytosolic caspase-2. The fatty acids also increased nitric oxide production, apparently independently of inducible nitric oxide synthase induction. Under the same experimental conditions, elevated glucose alone had only a marginal effect on beta-cell apoptosis. Together the results indicate that elevated concentrations of fatty acids are particularly efficient in accelerating the rate of apoptosis of already stressed beta(INS-1)-cells displaying altered mitochondrial function, and that the mitochondrial arm of the apoptosis process is involved in beta-cell lipotoxicity.

Apoptosis↗

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

Direct visualization by confocal fluorescent microscopy of the permeation of myristoylated peptides through the cell membrane.

To study the permeability through the cellular membrane of synthetic peptides containing an hydrophobic moiety, we used a 13-mer myristoylated peptide labeled with a N-terminal fluorescent probe. After 2 h of incubation, the subcellular distribution was analyzed in intact chromaffin cells by confocal fluorescent microscopy. Our results demonstrate that myristoylated peptides diffuse into intact cells, showing an heterogeneous distribution, but they do not reach the cellular nucleous, at least during the time range used.

Acylation↗

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↗

PubMed1991

Inactivation of Delayed Potassium Current in Cultured Bovine Chromaffin Cells.

Potassium currents of cultured bovine chromaffin cells in the presence of CdCl2 (0.5 mM) or CoCl2 (2 - 4 mM) may be described as a component that follows an n4 kinetics over the first 100 ms. However, during prolonged voltage clamp steps (5 - 20 s) this current reaches a peak and then declines to a non-zero steady-state level. In the absence of potassium accumulation, slow decay has been attributed to slow inactivation of the K+ conductance. Slow inactivation of calcium-independent potassium currents responds to the following: (i) the inactivation has a time course that may be fitted by a sum of two exponentials; (ii) inactivation is use-dependent (increasing frequency or duration of depolarizing pulses increases the extent of inactivation). Use-dependent inactivation does not appear when short (100 ms) hyperpolarizing prepulses are applied; (iii) at a membrane potential of - 60 mV, approximately 20% of the channels are inactivated; (iv) recovery from inactivation is very slow. At a holding potential of - 70 mV it takes about 20 s to reach the original value. The rate of inactivation recovery is voltage-dependent. The more hyperpolarized the membrane potential, the faster the recovery. The presence of use-dependent inactivation with slow recovery could be relevant for the stimulus - secretion coupling modulation in chromaffin cells.

Journal Article↗