Calcium-dependent Current Research Articles

Wnt-5a/Ca2+ pathway modulates endogenous current and oocyte structure of Xenopus laevis

Wnt signaling plays an essential role in cellular processes like development, maturation, and function maintenance. Xenopus laevis oocytes are a suitable model to study not only the development but also the function of different receptors expressed in their membranes, like those receptors expressed in the central nervous system (CNS) including Frizzled 7. Here, using frog oocytes and recordings of endogenous membrane currents in a two-electrode path configuration along with morphological observations, we evaluated the role of the non-canonical Wnt-5a ligand in oocytes. We found that acute application of Wnt-5a generated changes in endogenous calcium-dependent currents, entry oscillatory current, the membrane's outward current, and induced membrane depolarization. The incubation of oocytes with Wnt-5a caused a reduction of the membrane potential, potassium outward current, and protected the ATP current in the epithelium/theca removed (ETR) model. The oocytes exposed to Wnt-5a showed increased viability and an increase in the percentage of the germinal vesicle breakdown (GVBD), at a higher level than the control with progesterone. Altogether, our results suggest that Wnt-5a modulates different aspects of oocyte structure and generates calcium-dependent endogenous current alteration and GVDB process with a change in membrane potential at different concentrations and times of the exposition. These results help to understand the cellular effect of Wnt-5a and present the use of Xenopus oocytes to explore the mechanism that could impact the activation of Wnt signaling.

Calcium dependent current recordings in Xenopus laevis oocytes in microgravity

Mechanical unloading by microgravity (or weightlessness) conditions triggers profound adaptation processes at the cellular and organ levels. Among other mechanisms, mechanosensitive ion channels are thought to play a key role in allowing cells to transduce mechanical forces. Previous experiments performed under microgravity have shown that gravity affects the gating properties of ion channels. Here, a method is described to record a calcium-dependent current in native Xenopus laevis oocytes under microgravity conditions during a parabolic flight. A 3-voltage-step protocol was applied to provoke a calcium-dependent current. This current increased with extracellular calcium concentration and could be reduced by applying extracellular gadolinium. The custom-made “OoClamp” hardware was validated by comparing the results of the 3-voltage-step protocol to results obtained with a well-established two-electrode voltage clamp (TEVC). In the context of the 2nd Swiss Parabolic Flight Campaign, we tested the OoClamp and the method. The setup and experiment protocol worked well in parabolic flight. A tendency that the calcium-dependent current was smaller under microgravity than under 1 g condition could be observed. However, a conclusive statement was not possible due to the small size of the data base that could be gathered.

Adaptation in the visual cortex: influence of membrane trajectory and neuronal firing pattern on slow afterpotentials.

The input/output relationship in primary visual cortex neurons is influenced by the history of the preceding activity. To understand the impact that membrane potential trajectory and firing pattern has on the activation of slow conductances in cortical neurons we compared the afterpotentials that followed responses to different stimuli evoking similar numbers of action potentials. In particular, we compared afterpotentials following the intracellular injection of either square or sinusoidal currents lasting 20 seconds. Both stimuli were intracellular surrogates of different neuronal responses to prolonged visual stimulation. Recordings from 99 neurons in slices of visual cortex revealed that for stimuli evoking an equivalent number of spikes, sinusoidal current injection activated a slow afterhyperpolarization of significantly larger amplitude (8.5±3.3 mV) and duration (33±17 s) than that evoked by a square pulse (6.4±3.7 mV, 28±17 s; p<0.05). Spike frequency adaptation had a faster time course and was larger during plateau (square pulse) than during intermittent (sinusoidal) depolarizations. Similar results were obtained in 17 neurons intracellularly recorded from the visual cortex in vivo. The differences in the afterpotentials evoked with both protocols were abolished by removing calcium from the extracellular medium or by application of the L-type calcium channel blocker nifedipine, suggesting that the activation of a calcium-dependent current is at the base of this afterpotential difference. These findings suggest that not only the spikes, but the membrane potential values and firing patterns evoked by a particular stimulation protocol determine the responses to any subsequent incoming input in a time window that spans for tens of seconds to even minutes.

Voltage-Dependent Calcium Channels at the Plasma Membrane, but Not Vesicular Channels, Couple Exocytosis to Endocytosis

Although calcium influx triggers endocytosis at many synapses and non-neuronal secretory cells, the identity of the calcium channel is unclear. The plasma membrane voltage-dependent calcium channel (VDCC) is a candidate, and it was recently proposed that exocytosis transiently inserts vesicular calcium channels at the plasma membrane, thus triggering endocytosis and coupling it to exocytosis, a mechanism suggested to be conserved from sea urchin to human. Here, we report that the vesicular membrane, when inserted into the plasma membrane upon exocytosis, does not generate a calcium current or calcium increase at a mammalian nerve terminal. Instead, VDCCs at the plasma membrane, including the P/Q-type, provide the calcium influx to trigger rapid and slow endocytosis and, thus, couple endocytosis to exocytosis. These findings call for reconsideration of the vesicular calcium channel hypothesis. They are likely to apply to many synapses and non-neuronal cells in which VDCCs control exocytosis, and exocytosis is coupled to endocytosis.

Mutation in the myelin proteolipid protein gene alters BK and SK channel function in the caudal medulla

Proteolipid protein (Plp) gene mutation in rodents causes severe CNS dysmyelination, early death, and lethal hypoxic ventilatory depression (Miller et al., 2004). To determine if Plp mutation alters neuronal function critical for control of breathing, the nucleus tractus solitarii (nTS) of four rodent strains were studied: myelin deficient rats (MD), myelin synthesis deficient (Plp(msd)), and Plp(null) mice, as well as shiverer (Mbp(shi)) mice, a myelin basic protein mutant. Current-voltage relationships were analyzed using whole-cell patch-clamp in 300 microm brainstem slices. Voltage steps were applied, and inward and outward currents quantified. MD, Plp(msd), and Plp(null), but not Mbp(shi) neurons exhibited reduced outward current in nTS at P21. Apamin blockade of SK calcium-dependent currents and iberiotoxin blockade of BK calcium-dependent currents in the P21 MD rat demonstrated reduced outward current due to dysfunction of these channels. These results provide evidence that Plp mutation specifically alters neuronal excitability through calcium-dependent potassium channels in nTS.

Hyped up about the hypothalamus

Hyped up about the hypothalamus

Simulation analysis of bandpass filtering properties of a rod photoreceptor network

The bandpass filtering properties of a rod network were studied via computer simulations. Sinusoidal current stimuli were applied to a single rod model to characterize its temporal filtering properties. The simulated frequency response revealed that a single rod behaves as a bandpass filter whose characteristics are affected by the stimulus strength and frequency. We analyzed the contribution of individual ionic currents to bandpass filtering and found that the filtering of small signals is largely regulated by the calcium-dependent currents I K(Ca) and I Cl(Ca), whereas the filtering of large signals is regulated by the hyperpolarization-activated current, I h . Furthermore, rod network modeling by electrically interconnecting the single rod models revealed that the acceleration of signals that spread laterally through the rod network is attributed to I K(Ca) and not I h .

Modeling and simulation of retinal neurons and the neuroinformatics tools

Event Abstract Back to Event Modeling and simulation of retinal neurons and the neuroinformatics tools Yoshimi Kamiyama1* and Shiro Usui2 1 Aichi Prefectural University, Japan 2 RIKEN Brain Science Institute, Japan The retina transforms the light into nerve spikes which are carried by the optic nerve to the brain. The retina has been thought to be a window to the brain because of its accessibility and suitability for neuroscientific investigation. It is thought to be one of the few parts of the vertebrate brain where we can reasonably explain its purpose and how it works. The retina is an attractive model system for approaching the question of the information processing role of biological mechanisms of neurons. The physiology and anatomy of the retina are relatively well known. Physiological studies of the retina have uncovered a number of cellular and subcellular mechanisms such as phototransduction and the characteristics of the ion channels found in retinal neurons. Neuroanatomical studies have revealed the morphological principles govering the structure of the retina. The former data provide information about the functional role of ion channels in generating and shaping the light response of the retinal neurons. It is as yet impossible to provide a complete model of the retina. But it is now possible to identify some computational operations that the retina perfoms and to relate them to specific physiological mechanisms, on the basis of computational modeling work. Over the last decade, we have developed mathematical models of retinal neurons based on neurophysiological details. These models can reveal what happens if a particular ionic current is active or inactive. One can then ask what advantages are given on the visual system by having the particular class of ion channel present in the neuron. Here, we show the mathematical models of the retinal neurons, and demonstrate that the electircal responses are quantitatively accounted for by the models. The models we have developed provide a unified framework for the undestanding of the functional role of cellular mechanisms of retinal neurons and their integrative functions. In the present study, we analyzed the underlying mechanisms of a rod photoreceptor that behaves as a bandpass filter which has long been a mystery. We discovered that the kinetics of the calcium dependent ionic currents characterize the bandpass filtering of rod signals. We also developed a stochastic model of spike generation in retinal ganglion cells. The stochastic model showed precise and reliable spikes to randomly fluctuating current similar to those of experimental data. We found the spike timing was much influenced by the outward currents, not by the sodium current which riggers spikes. The simulation analysis with these models requires some software tools. We developed three software systems; (1) XML-based retinal neuron modeling language which stores not only mathematical models, but also all calculation conditions and improves model reproducibility, (2)Grid portal simulation environment which provides seamless access to computing resources over the network, (3)Realtime retinal neuron simulator with USB camera device for understanding the role of retinal neurons in visual information processing. These tools highly improved simulation efficiency for retinal modeling and simulation. 81b tn_07-29-2008-10-52_81b tn_81b 07-29-2008-10-52_81b Conference: Neuroinformatics 2008, Stockholm, Sweden, 7 Sep - 9 Sep, 2008. Presentation Type: Oral Presentation Topic: Live Demonstrations Citation: Kamiyama Y and Usui S (2008). Modeling and simulation of retinal neurons and the neuroinformatics tools. Front. Neuroinform. Conference Abstract: Neuroinformatics 2008. doi: 10.3389/conf.neuro.11.2008.01.121 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 28 Jul 2008; Published Online: 28 Jul 2008. * Correspondence: Yoshimi Kamiyama, Aichi Prefectural University, Aichi, Japan, usuishiro@riken.jp Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract Supplemental Data The Authors in Frontiers Yoshimi Kamiyama Shiro Usui Google Yoshimi Kamiyama Shiro Usui Google Scholar Yoshimi Kamiyama Shiro Usui PubMed Yoshimi Kamiyama Shiro Usui Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Cell Swelling Triggers Glutamate Release

Astrocytes are becoming recognized as active participants in modulating neuronal activity. Indeed, astrocytes accumulate glutamate and release this neurotransmitter in response to purinergic receptor activation by ATP. Takano et al. provide evidence that ATP triggers transient cell swelling, which activates volume-sensitive channels, allowing glutamate and other osmolytes, such as aspartate and taurine, to be released. Cell swelling was detected in three different assays. In the presence of hypertonic medium, ATP-stimulated release of glutamate was abolished. Release of glutamate, taurine, and aspartate also required increased calcium concentrations and was blocked by calcium chelators. Whole-cell currents were measured in cultured astrocytes and showed an ATP-stimulated inward current mediated by glutamate release (under conditions where glutamate was the only possible charge carrier). Finally, a similar ATP-stimulated and calcium-dependent glutamate current was recorded in astrocytes in hippocampal slices using two-photon microscopy and whole-cell current clamp. Thus, glutamate and other osmolyte amino acids appear to be released from astrocytes by a volume-sensitive channel that is activated in response to cell swelling caused by activation of purinergic receptors. The strongest evidence against calcium-dependent vesicular release is the inhibition of release in the presence of hypertonic medium, which typically stimulates vesicular release. T. Takano, J. Kang, J. K. Jaiswal, S. M. Simon, J. H.-C. Lin, Y. Yu, Y. Li, J. Yang, G. Dienel, H. R. Zielke, M. Nedergaard, Receptor-mediated glutamate release from volume sensitive channels in astrocytes. Proc. Natl. Acad. Sci. U.S.A. 102 , 16466-16471 (2005). [Abstract] [Full Text]

Nitric oxide and histamine induce neuronal excitability by blocking background currents in neuron MCC of Aplysia.

Nitric oxide (NO) and histamine are important neurotransmitters and neuromodulators. We investigated their ability to modulate the membrane ionic currents and excitability of the metacerebral cell (MCC) of Aplysia using voltage clamp techniques. MCC is a serotonergic modulator of the feeding neural circuit. It receives powerful long-lasting excitatory synaptic input mediated by NO and histamine. NO donors reduced a background outward current at and above the resting potential, associated with decreased membrane conductance. This produced a substantial steady-state inward current that was relatively insensitive to cesium or cobalt. The NO response appears to be due to the reduction of a background potassium current and a small increase in persistent inward sodium current. Treatment with 8-bromoguanosine-3'5'-cyclic monophosphate mimics this response, suggesting it is mediated primarily by the NO-guanylyl cyclase-cGMP pathway. In some MCCs, NO blocked an additional potassium current that resulted in current reversal near the potassium equilibrium potential in current-voltage plots. Histamine also reduced a background outward current at and above the resting potential. However, treatment with cobalt, which blocks calcium and calcium-dependent currents, blocked the histamine response, suggesting that histamine decreases calcium activated potassium currents. Although nifedipine (L-type calcium channel blocker) and tetraethylammonium reduced some calcium and calcium-dependent potassium currents, they had only a slight effect on the NO and histamine responses. Both NO and histamine decreased steady-state membrane currents, and thereby depolarized MCC and increased its excitability, but different ionic currents and second messenger pathways are involved, allowing complex state and time dependent modulation of MCC's activity.

Temporal integration by calcium dynamics in a model neuron.

The calculation and memory of position variables by temporal integration of velocity signals is essential for posture, the vestibulo-ocular reflex (VOR) and navigation. Integrator neurons exhibit persistent firing at multiple rates, which represent the values of memorized position variables. A widespread hypothesis is that temporal integration is the outcome of reverberating feedback loops within recurrent networks, but this hypothesis has not been proven experimentally. Here we present a single-cell model of a neural integrator. The nonlinear dynamics of calcium gives rise to propagating calcium wave-fronts along dendritic processes. The wave-front velocity is modulated by synaptic inputs such that the front location covaries with the temporal sum of its previous inputs. Calcium-dependent currents convert this information into concomitant persistent firing. Calcium dynamics in single neurons could thus be the physiological basis of the graded persistent activity and temporal integration observed in neurons during analog memory tasks.

Regulation of firing response gain by calcium-dependent mechanisms in vestibular nucleus neurons.

Behavioral reflexes can be modified by experience via mechanisms that are largely unknown. Within the circuitry for the vestibuloocular reflex (VOR), neurons in the medial vestibular nucleus (MVN) show adaptive changes in firing rate responses that are correlated with VOR gain (the ratio of evoked eye velocity to input head velocity). Although changes in synaptic strength are typically assumed to underlie gain changes in the VOR, modulation of intrinsic ion channels that dictate firing could also play a role. Little is known, however, about how ion channel function or regulation contributes to firing responses in MVN neurons. This study examined contributions of calcium-dependent currents to firing responses in MVN neurons recorded with whole cell patch electrodes in rodent brain stem slices. Firing responses were remarkably linear over a wide range of firing rates and showed modest spike frequency adaptation. Firing response gain, the ratio of evoked firing rate to input current, was reduced by increasing extracellular calcium and increased either by lowering extracellular calcium or with antagonists to SK- and BK-type calcium-dependent potassium channels and N- and T-type calcium channels. Blockade of SK channels occluded gain increases via N-type calcium channels, while blocking BK channels occluded gain increases via presumed T-type calcium channels, indicating specific coupling of potassium channels and their calcium sources. Selective inhibition of Ca(2+)/calmodulin-dependent kinase II and broad-spectrum inhibition of phosphatases modulated gain via BK-dependent pathways, indicating that firing responses are tightly regulated. Modulation of firing response gain by phosphorylation provides an attractive mechanism for adaptive control of VOR gain.

How is bursting mode maintained under the action of exo- and endogenous factors?

The investigation is concerned with slow oscillations of the intracellular calcium concentration, fast oscillations of the membrane potential, and their interactions during bursting generation. The question associated with functioning of the bursting mode is considered: how is the condition of simultaneous existence of the slow and fast membrane potential oscillations (necessary for the bursting mode) realized under exo- and endogenous actions that result in changes in the neuron membrane parameters? To answer the question the regions of existence of fast, slow and bursting oscillations have been constructed on different parameter planes. The study has revealed that in spite of the fact the regions of the existence of the slow and fast oscillations are sensitive to parameter variations, the condition of the bursting mode existence remains due to invariance of the spatial vicinity of these regions. Furthermore, it has been demonstrated that there exists the possibility to enhance the maximal conductances of the calcium-dependent currents simultaneously such that even strong variations of their values do not induce changes of the oscillations mode (i.e. it remains bursting).

Calcium channels in Xenopus spinal neurons differ in somas and presynaptic terminals.

Calcium channels play dual roles in cell signaling by promoting membrane depolarization and allowing entry of calcium ions. Patch-clamp recordings of calcium and calcium-dependent currents from the soma of Xenopus spinal neurons indicate key functional differences from those of presynaptic terminals. Both terminals and somas exhibit prominent high-voltage-activated (HVA) calcium current, but only the soma expresses additional low-voltage-activated (LVA) T-type current. Further differences are reflected in the HVA current; N- and R-type channels are predominant in the soma while the terminal calcium current is composed principally of N type with smaller contribution by L- and R-type channels. Potential physiological significance for these different distributions of channel types may lie in the differential channel kinetics. Activation of somatic HVA calcium current occurs more slowly than HVA currents in terminals. Additionally, somatic LVA calcium current activates and deactivates much more slowly than any HVA calcium current. Fast-activating and -deactivating calcium current may be critical to processing the rapid exocytotic response in terminals, whereas slow LVA and HVA calcium currents may play a central role in shaping the somatic firing pattern. In support of different kinetic behavior between these two compartments, we find that somatic calcium current activates a prominent slow chloride current not observed in terminal recordings. This current activates in response to calcium entering through either LVA or HVA channels and likely functions as a modulator of excitability or synaptic input. The restriction of this channel type to the soma lends further support to the idea that differential expression of fast and slow channel types in these neurons is dictated by differences in signaling requirements for somatic and terminal compartments.

Ion channel activity drives ion channel expression.

Ion channel activity drives ion channel expression.

Effects of hypertrophy on regional action potential characteristics in the rat left ventricle: a cellular basis for T-wave inversion?

In cardiac hypertrophy, ECG T-wave changes imply an abnormal sequence of ventricular repolarization. We investigated the hypothesis that this is due to changes in the normal regional differences in action potential duration. We assessed the contribution of potassium- and calcium-dependent currents to these differences. Both the altered sequence of ventricular repolarization and the underlying cellular mechanisms may contribute to the increased incidence of ventricular arrhythmias in hypertrophy. Rats received daily isoproterenol injections for 7 days. Myocytes were isolated from basal subendocardial (endo), basal midmyocardial (mid), and apical subepicardial (epi) regions of the left ventricular free wall. Action potentials were stimulated with patch pipettes at 37 degrees C. The ratio of heart weight to body weight and mean cell capacitance are increased by 22% and 18%, respectively, in hypertrophy compared with controls (P<.001). Normal regional differences in action potential duration at 25% repolarization (APD25) are reduced in hypertrophy (control: endo, 11.4+/-0.9 ms; mid, 8.2+/-0.9 ms; epi, 5.1+/-0.4 ms; hypertrophy: endo, 11.6+/-0.9 ms; mid, 10.4+/-0.8 ms; epi, 7.8+/-0.6 ms). The regional differences in APD25 are still present in 3 mmol/L 4-aminopyridine. Hypertrophy affects APD75 differently, depending on the region of origin of myocytes (ANOVA P<.05). APD75 is shortened in subendocardial myocytes but is prolonged in subepicardial myocytes (control: endo, 126+/-7 ms; epi, 96+/-10 ms; hypertrophy: endo, 91+/-6 ms; epi, 108+/-7 ms). These changes in APD75 are altered by intracellular calcium buffering. Normal regional differences in APD and the changes observed in hypertrophy are only partially explained by differences in I(tol). In hypertrophy, the normal endocardial/epicardial gradient in APD75 appears to be reversed. This may explain the T-wave inversion observed and will have implications for arrhythmogenesis.

Calcium release from intracellular stores evoked by extracellular ATP in a Xenopus renal epithelial cell line.

1. The signal transduction mechanism mediating extracellular adenosine 5'-triphosphate (ATP)-induced calcium release in a renal epithelial cell line (A6) was investigated using the whole-cell voltage-clamp technique and fura-2 fluorescence measurement. 2. ATP (10 microM) activated calcium-dependent non-selective cation channels in cells held under voltage clamp. 3. Guanosine 5'-O-(2-thiodiphosphate) (GDP beta S; 0.1-1.0 mM) in the pipette inhibited the ATP-activated calcium-dependent currents. With guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S; 0.1-1.0 mM) in the pipette, the currents were spontaneously elicited without application of ATP. Pretreatment with pertussis toxin (PTX) affected neither the ATP-activated currents nor the increase in intracellular free calcium concentration ([Ca2+]i) evoked by ATP. 4. Intracellular application of neomycin or heparin inhibited the ATP-activated currents. Inositol 1,4,5-trisphosphate (IP3; 0.1-100 microM) in the internal solution produced currents similar to those due to ATP activation. 5. These results suggest that a PTX-insensitive guanosine 5'-triphosphate (GTP)-binding regulatory protein (G. protein) is involved in extracellular. ATP-induced phosphoinositide turnover and subsequent calcium release from IP3-sensitive stores, which subsequently activates the calcium-dependent channels in A6 cells.

Potassium currents in isolated CA1 neurons of the rat after kindling epileptogenesis

Daily tetanic stimulation of the Schaffer collaterals generates an epileptogenic focus in area CA1 of the rat hippocampus, ultimately leading to generalized tonic-clonic convulsions (kindling). Potassium currents were measured under voltage-clamp conditions in pyramidal neurons, acutely dissociated from the focus of fully kindled rats, one day and six weeks after the last generalized seizure. Their amplitude, kinetics, voltage dependence and calcium dependence were compared with controls. With Ca2+ influx blocked by 0.5 mM Ni2+, the sustained current (delayed rectifier) and the transient current (A-current) were not different after kindling. Calcium influx evoked an additional fast transient current component. This transient calcium-dependent current component was increased by 154%, but only immediately after the seizure. A second, slow calcium-dependent potassium current component was dependent on the intracellular calcium level, set by the pipette as well as on calcium influx. The peak amplitude of this slow calcium-dependent current was under optimal calcium conditions not different after kindling, but we found indications that either calcium homeostasis or the calcium sensitivity of the potassium channels was affected by the kindling process. In contrast to the previously described enhancement of calcium current, kindling epileptogenesis did not change the total potassium current amplitude. The minor changes that were observed can be related either to changes in calcium current or to changes in intracellular calcium homeostasis.

Nitric oxide as a nonadrenergic inhibitory transmitter in smooth muscle cells of the guinea pig gastro-intestinal tract: Mechanisms of action

The role of nitric oxide (NO) as a possible transmitter for nonadrenergic inhibitory transmission was studied on isolated muscle strips of the guinea pig gastro-intestinal tract (GIT) using sucrose-gap technique. In addition, the voltage clamp and intracellular dialysis techniques were employed to study the effects of sodium nitroprusside (NP) on isolated smooth muscle (SM) cells of thetaenia coli. N-nitro-L-arginine methyl ester (L-NAME), a blocker of NO synthesis from L-arginine (0.1 mM), was shown to selectively suppress the apamin-resistant component of nonadrenergic inhibitory junctional (synaptic) potentials (IJP) in the guinea pig GIT SM cells. At the same time, L-NAME did not affect the vasoactive intestinal polypeptide (VIP)- and NP-evoked hyperpolarization in SM cells of the colon. The NP-induced hyperpolarization (0.1 mM) was accompained by a decrease in the SM cell membrane resistance. Application of NP to isolated SM cells activated a small outward current and increased the frequency of spontaneous transient calcium-dependent outward currents. NP increased the Ca-dependent potassium current evoked in SM cells by step depolarization, but did not affect the potassium currents of delayed rectification. Our results suggest that NO is involved in generation of nonadrenergic IJP in SM cells of the guinea pig GIT. The action of NP on SM cells is complex and results in hyperpolarization and relaxation (partially through the activation of Ca-dependent potassium channels in SM cell membrane).

A branching dendritic model of a rodent CA3 pyramidal neurone.

1. We constructed a branching dendritic compartmental model of a CA3 pyramidal neurone, using experimental data from guinea-pig and rat cells obtained in vitro. The goal was to understand interactions between synaptic events impinging on dendritic branches and voltage- and calcium-dependent currents. The model contained sixty-four soma-dendrite (SD) compartments, an axon initial segment (IS), and four axonal compartments. There were six active conductances in the SD membrane, including a sodium conductance (gNa) and a high-threshold calcium conductance (gCa), with kinetic properties similar to those reported in a previous study. 2. The distribution of conductance densities across the IS and SD was adjusted by testing the model response to antidromic stimulation and current pulses or sustained currents injected into the soma or apical dendrites. As before, gNa was concentrated on and near the soma with lower density in the dendrites, while gCa had a higher density in apical dendrites than at the soma. 3. The model predicts that CA3 pyramidal neurones in media blocking synaptic transmission should fire a burst of action potentials following antidromic stimulation. This was confirmed experimentally in hippocampal slices. 4. Both in the model and in guinea-pig neurones, dendritic IPSCs can delay the onset of bursting. If an IPSC begins soon enough after the first fast action potential, the later burst envelope is attenuated. This effect results from suppression of dendritic Ca2+ electrogenesis. 5. The model predicts that an appropriately timed dendritic IPSC (after the first somatic spike but before the dendritic Ca2+ spike) may suppress the transient local [Ca2+] signal, while having a negligible effect on the electrical output of the neurone. This phenomenon has been reported in guinea-pig Purkinje cells. 6. We conclude that active dendritic currents are critical for regulation of the electrical output of CA3 pyramidal neurones. We suggest also that dendritic [Ca2+] signals might be controlled in individual dendrites independently of action potential outputs, an effect of possible importance for synaptic plasticity.