EFhd2/Swiprosin-1 is a common genetic determinator for sensation-seeking/low anxiety and alcohol addiction.

In many societies, the majority of adults regularly consume alcohol. However, only a small proportion develops alcohol addiction. Individuals at risk often show a high sensation-seeking/low-anxiety behavioural phenotype. Here we asked which role EF hand domain containing 2 (EFhd2; Swiprosin-1) plays in the control of alcohol addiction-associated behaviours. EFhd2 knockout (KO) mice drink more alcohol than controls and spontaneously escalate their consumption. This coincided with a sensation-seeking and low-anxiety phenotype. A reversal of the behavioural phenotype with ß-carboline, an anxiogenic inverse benzodiazepine receptor agonist, normalized alcohol preference in EFhd2 KO mice, demonstrating an EFhd2-driven relationship between personality traits and alcohol preference. These findings were confirmed in a human sample where we observed a positive association of the EFhd2 single-nucleotide polymorphism rs112146896 with lifetime drinking and a negative association with anxiety in healthy adolescents. The lack of EFhd2 reduced extracellular dopamine levels in the brain, but enhanced responses to alcohol. In confirmation, gene expression analysis revealed reduced tyrosine hydroxylase expression and the regulation of genes involved in cortex development, Eomes and Pax6, in EFhd2 KO cortices. These findings were corroborated in Xenopus tadpoles by EFhd2 knockdown. Magnetic resonance imaging (MRI) in mice showed that a lack of EFhd2 reduces cortical volume in adults. Moreover, human MRI confirmed the negative association between lifetime alcohol drinking and superior frontal gyrus volume. We propose that EFhd2 is a conserved resilience factor against alcohol consumption and its escalation, working through Pax6/Eomes. Reduced EFhd2 function induces high-risk personality traits of sensation-seeking/low anxiety associated with enhanced alcohol consumption, which may be related to cortex function.

Activin tunes GABAergic neurotransmission and modulates anxiety-like behavior.

Activin, a member of the transforming growth factor-beta superfamily, affords neuroprotection in acute brain injury, but its physiological functions in normal adult brain are largely unknown. Using transgenic (tg) mice expressing a dominant-negative activin receptor mutant under the control of the CaMKIIalpha promoter in forebrain neurons, we identified activin as a key regulator of gamma-aminobutyric acid (GABA)ergic synapses and anxiety-like behavior. In the open field, wild-type (wt) and tg mice did not differ in spontaneous locomotion and exploration behavior. However, tg mice visited inner fields significantly more often than wt mice. In the light-dark exploration test, tg mice made more exits, spent significantly more time on a well-lit elevated bar and went farther away from the dark box as compared to wt mice. In addition, the anxiolytic effect of diazepam was abrogated in tg mice. Thus the disruption of activin receptor signaling produced a low-anxiety phenotype that failed to respond to benzodiazepines. In whole-cell recordings from hippocampal pyramidal cells, enhanced spontaneous GABA release, increased GABA tonus, reduced benzodiazepine sensitivity and augmented GABA(B) receptor function emerged as likely substrates of the low-anxiety phenotype. These data provide strong evidence that activin influences pre- and postsynaptic components of GABAergic synapses in a highly synergistic fashion. Given the crucial role of GABAergic neurotransmission in emotional states, anxiety and depression, dysfunctions of activin receptor signaling could be involved in affective disorders: and drugs affecting this pathway might show promise for psychopharmacological treatment.

Muscarinic activation of inwardly rectifying K(+) conductance reduces EPSPs in rat hippocampal CA1 pyramidal cells.

1. To determine how acetylcholine (ACh) modulates the somatodendritic processing of EPSPs, we performed whole-cell recordings from CA1 pyramidal cells of hippocampal slices and examined the effect of the cholinergic agonist, carbachol (CCh), on alpha-amino-3-hydroxy-5-methyl isoxazole-4-propionate (AMPA) EPSPs, miniature EPSPs, and EPSP-like waveforms evoked by brief dendritic glutamate pulses (glutamate-evoked postsynaptic potentials, GPSPs). 2. Although CCh is known to enhance the intrinsic excitability of the neuron in several ways, activation of atropine-sensitive (muscarinic) receptors on the apical dendrite or the soma of CA1 pyramidal cells consistently reduced the amplitude of EPSPs and GPSPs. 3. Cholinergic inhibition of evoked and simulated EPSP waveforms displayed considerable voltage dependence, with the amplitude of the postsynaptic potentials progressively declining with membrane hyperpolarization indicating the involvement of an inwardly rectifying current. 4. Extracellular Ba(2+) (200 microM) and tertiapin (30 nM), a novel and selective blocker of G protein-activated, inwardly rectifying K(+) (GIRK) channels, completely blocked the effect of CCh on GPSP amplitude. 5. Muscarinic reduction of GPSPs was not sensitive to the M1 receptor-preferring antagonist, pirenzepine, but was suppressed by the M2 receptor-preferring antagonist, methoctramine, and by the allosteric M2 receptor antagonist, gallamine. 6. In voltage-clamp recordings, CCh induced an ion current displaying inward rectification in the hyperpolarizing direction, which was identified as a GIRK current based on its sensitivity to low Ba(2+) and tertiapin. Its pharmacological profile paralleled that of the cholinergic GPSP reduction. 7. We link the observed reduction of postsynaptic potentials to the cholinergic activation of a GIRK conductance, which serves to partially shunt excitatory synaptic input.

The roles of activins in repair processes of the skin and the brain.

A recent study from our laboratory demonstrated a strong upregulation of activin expression during cutaneous wound healing. To further analyze the role of activin A in skin morphogenesis and wound repair, we generated transgenic mice that overexpress activin A under the control of the keratin 14 promoter. The latter targets expression of transgenes to the basal, proliferating layer of the epidermis. Hetero- as well as homozygous transgenic animals were viable and fertile. However, they were smaller than non-transgenic littermates and they had smaller ears and shorter tails. Histological analysis of their skin revealed dermal hyperthickening, mainly due to the replacement of fatty tissue by connective tissue, and an increase in suprabasal, partially differentiated epidermal layers. After cutaneous injury, a strong enhancement of granulation tissue formation was observed. Furthermore, the extent of re-epithelialization was increased in some of the wounds. These data demonstrate that activin A is a potent stimulator of the wound healing process. Using an in vivo model of local brain injury, we found that activin A also plays a significant role in the early cellular response to neuronal damage. Expression of activin mRNA and protein is markedly upregulated within a few hours of injury. If applied exogenously, recombinant activin A is capable of rescuing neurons from acute cell death. Studying the interaction between bFGF, a well-established neuroprotective agent, which is currently being tested in stroke patients, and activin A, we arrived at the unexpected conclusion that it is the strong induction of activin A by bFGF which endows the latter with its beneficial actions in patients. These findings suggest that the development of substances directly targeting activin expression or receptor binding should offer new possibilities in the acute treatment of stroke and brain trauma.

GABA-evoked chloride currents do not differ between dendrites and somata of rat neocortical neurons.

1. We performed patch-clamp recordings on acutely isolated somata and dendritic segments of rat neocortical neurons, in order to compare the reversal potential (E(GABA)) and relative density of GABA(A) receptor-mediated Cl(-) currents in these two cellular compartments. 2. Currents were recorded with the Cl(-)-impermeable pore former gramicidin (25--75 microg ml(-1)) in HCO(3)(-)-free bath solution. Voltage ramps (-110 to -30 mV) from a holding potential (V(h)) of -60 mV in the absence and presence of 2 microM GABA were used to construct instantaneous current-voltage relationships. Currents were abolished by co-application of GABA with the GABA(A) receptor antagonist bicuculline (40 microM). 3. GABA conductance, normalized to membrane surface area, was not different in somata and dendrites. In addition, E(GABA) was not different in the two compartments. 4. Replacement of intracellular K(+) with Cs(+) resulted in a significantly more depolarized E(GABA) in both somata and dendrites. These results suggest that the resting intracellular Cl(-) concentration ([Cl(-)](i)) is similar in somata and dendrites and that an outward Cl(-) transporter system maintains low [Cl(-)](i).

Expression and biological significance of Ca2+-activated ion channels in human keratinocytes.

In whole-cell recordings from HaCaT keratinocytes, ATP, bradykinin, and histamine caused a biphasic change of the membrane potential consisting of an initial transient depolarization, followed by a pronounced and long-lasting hyperpolarization. Flash photolysis of caged IP3 mimicked the agonist-induced voltage response, suggesting that intracellular Ca2+ release and subsequent opening of Ca2+-activated ion channels serve as the common transduction mechanism. In contrast, cAMP- and PKC-dependent pathways were not involved in the electrophysiological effects of the extracellular signaling molecules. The depolarization was predominantly mediated by a DIDS- and niflumic acid-sensitive Cl- current, whereas a charybdotoxin- and clotrimazole-sensitive K+ current underlay the prominent hyperpolarization. Consistent with the electrophysiological data, RT-PCR showed that HaCaT keratinocytes express two types of Ca2+-activated Cl- channels, CaCC2 and CaCC3 (CLCA2), as well as the Ca2+-activated K+ channel hSK4. That the pronounced hSK4-mediated hyperpolarization bears significance on the growth and differentiation properties of keratinocytes is suggested by RNase protection assays showing that hSK4 mRNA expression is strongly down-regulated under conditions that allow keratinocyte differentiation. hSK4 might thus play a role in linking changes in membrane potential to the biological fate of keratinocytes.

Enhancement of persistent Na+ current by sea anemone toxin (ATX II) exerts dual action on hippocampal excitability.

We used anemone toxin II (ATX II) to study how a selective enhancement of persistent Na+ current (INaP) would affect the excitability of CA1 pyramidal neurons in the hippocampal slice. In whole-cell recordings from CA1 cell somata, local application of ATX II (10 microM) into the stratum pyramidale invariably depolarized the neurons and produced sustained burst discharges with depolarizing plateau potentials of variable amplitude and length. However, the strong excitatory action of ATX II, observed on the single cell level, was not mirrored in field potential recordings from the same hippocampal subfield. The amplitude of the electrically evoked population spike declined, reflecting the decreased availability of fast Na+ channels, and the intracellulary recorded burst discharges were not detected by the field electrode. The lacking synchronization of cellular bursting activity was seen during both local and bath application of ATX II, suggesting that the toxin, in addition to promoting burst discharges of individual neurons, simultaneously dampens network excitability. In fact, ATX II reduced afferent fibre volleys (reflecting axonal excitability) and field excitatory postsynaptic potentials (EPSPs) in a similar fashion. As the expression of different Na+ channel subtypes appears to be compartmentalized within hippocampal neurons, we propose that point mutations leading to pathologically enhanced INaP might exert quite opposite effects, depending on the type and location of the Na+ channel affected. Whereas alterations of somatodendritic Na+ channels would give rise to bursting activity, alterations of axonal Na+ channels would primarily decrease network excitability.

Induction of activin A is essential for the neuroprotective action of basic fibroblast growth factor in vivo.

Exogenous application of neurotrophic growth factors has emerged as a new and particularly promising approach not only to promote functional recovery after acute brain injury but also to protect neurons against the immediate effect of the injury. Among the various growth factors and cytokines studied so far, the neuroprotective and neurotrophic profile of basic fibroblast growth factor (bFGF) is the best documented. Using an animal model of acute excitotoxic brain injury, we report here that the neuroprotective action of bFGF, which is now being tested in stroke patients, depends on the induction of activin A, a member of the transforming growth factor-beta superfamily. Our evidence for this previously unknown mechanism of action of bFGF is that bFGF strongly enhanced lesion-associated induction of activin A; in the presence of the activin-neutralizing protein follistatin, bFGF was no longer capable of rescuing neurons from excitotoxic death; and recombinant activin A exerted a neuroprotective effect by itself. Our data indicate that the development of substances influencing activin expression or receptor binding should offer new ways to fight neuronal loss in ischemic and traumatic brain injury.

Connective tissue growth factor: a novel player in tissue reorganization after brain injury?

Recent studies have suggested a role of connective tissue growth factor (CTGF) in repair processes of the skin as well as in various types of fibrotic disease. However, a function of this molecule in central nervous system (CNS) repair has not been demonstrated yet. In this study we analysed the temporal and spatial expression pattern of CTGF after unilateral kainic acid lesions of the hippocampal CA3 region in mice. We found a strong induction of CTGF mRNA and protein expression in neurons and glial cells of the lesioned hippocampus. Interestingly, increased expression of this mitogen was accompanied by elevated levels of the extracellular matrix molecule fibronectin, which is a known target of CTGF action. Therefore, our data indicate a novel function of CTGF in postlesional restructuring of the hippocampus, where it possibly participates in glial scar formation.

Variance analysis of current fluctuations of adenosine- and baclofen-activated GIRK channels in dissociated neocortical pyramidal cells.

Whole cell recordings were obtained from pyramidal cell somata acutely isolated from rat neocortex. In voltage-clamp mode, adenosine (0.3-1000 microM), and the GABA(B) receptor agonist, baclofen (1-300 microM), induced K+ current responses mediated by G protein-activated inwardly rectifying K+ (GIRK) channels. In our preparation, adenosine activated GIRK currents with an average EC(50) of 2 microM. Baclofen had an average EC50 of 26 microM. To estimate and compare unitary conductance and density of GIRK channels activated by either adenosine or baclofen, we performed variance analysis of current fluctuations associated with the application of the two agonists at increasing concentrations. Irrespective of the agonist tested, GIRK channels displayed an average single-channel conductance of 25 pS at our recording conditions ([K+]o: 60 mM). Assuming that GIRK channel conductance increases in proportion to the square root of [K+]o, this would translate into 5-6 pS at physiological ion gradients. GIRK channels activated by adenosine or baclofen were not only identical in terms of unitary conductance, they also displayed the same average density of 0.5 channels micron(-2) for both agonists. Our data strongly suggest that the two compounds recruit the same type of channel and thus most likely share a common transduction and effector system.

G protein-activated inwardly rectifying K+ (GIRK) currents in dendrites of rat neocortical pyramidal cells.

1. We performed patch-clamp recordings on acutely isolated dendritic segments and cell somata of rat neocortical pyramidal neurons to determine and compare the relative density of G protein-activated K+ (GIRK) currents in the two cellular compartments. 2. Hyperpolarizing voltage ramps and elevation of extracellular K+ concentration to 40 mM served to identify inwardly rectifying K+ currents. Near-saturating concentrations of adenosine (100 microM), baclofen (20 microM) and serotonin (20 microM) all produced robust GIRK currents in cell somata as well as in dendritic segments that were completely abolished by Ba2+ (200 microM). In addition to agonist-activated GIRK currents, both somata and dendrites displayed a constitutive Ba2+-sensitive inward rectification. 3. In order to compare the relative strengths of GIRK current responses in the two compartments, GIRK conductance was normalized to surface area. In contrast to intrinsic, G protein-independent inward rectification, which was comparable in size in the two compartments, all three agonists evoked significantly larger GIRK conductances in dendrites than in somata. 4. Our data suggest that several neurotransmitters might employ GIRK currents as a tool to directly modulate the electrical properties of dendrites. In concert with voltage-dependent K+ currents and the hyperpolarization-activated cation current (Ih) of the dendrite, GIRK currents should dampen dendritic excitability and thus influence various aspects of dendritic signal integration.

Activin: a novel player in tissue repair processes.

Recent studies have demonstrated a strong expression of activin in repair processes of various tissues and organs, including the skin, the lung, the intestine, the cardiovascular system, and even the brain. Although little is as yet known about the function of activin in tissue repair, first results suggest a role of activin in epithelial differentiation, fibroblast proliferation and expression of matrix molecules by these cells, and also in neuroprotection. Whereas a transient overexpression of activin after tissue injury might be beneficial for the repair process, sustained expression of activin could lead to fibrotic processes. Therefore, the modulation of the availability or biological activity of activin could be of particular importance for the treatment of impaired tissue repair on the one hand and tissue fibrosis on the other hand.

Spread of excitation in chronically lesioned mouse hippocampus determined by laser scanning microscopy.

Fast optical recordings by means of laser scanning microscopy in conjunction with a voltage-sensitive dye (RH 414) were performed to monitor the spatio-temporal spread of neuronal activity in CA3/CA4-lesioned C57BL6 mouse hippocampal slices prepared approximately 3 months after intracerebroventricular kainic acid (KA) injection. The aim of our study was to assess the effects of a circumscribed neuronal loss on the propagation of electrical activity along the trisynaptic hippocampal circuit. Both in physiological bathing solution and in bicuculline (10 microM), hilar stimulation failed to activate the downstream pathway, so that, under these conditions, the chronically disinhibited CA1 region appeared to be effectively isolated from burst activity arising upstream; however, epileptiform discharges evoked in zero Mg2+ solution were reliably transmitted from the dentate gyrus to the CA1 region. That these bursts were indeed spreading across the lesion, and not along newly formed connections (e.g., between dentate gyrus and CA1), was confirmed by acute transection experiments of the Schaffer collateral/commissural pathway, which completely abolished translesional burst propagation. The fact that the surviving CA3-CA1 connections are unable to trigger epileptiform bursts after suppression of GABAergic inhibition suggests that the lesioned region might serve as a filter that shields hyperexcitable CA1 neurons from epileptic activity arising upstream, in particular from chronically disinhibited granule cells of the dentate gyrus. An impaired GABAergic inhibition will thus only have minor facilitating effects on seizure propagation in the hippocampus of CA3-lesioned animals.

Muscarinic inhibition of persistent Na+ current in rat neocortical pyramidal neurons.

Muscarinic modulation of persistent Na+ current (INaP) was studied using whole cell recordings from acutely isolated pyramidal cells of rat neocortex. After suppression of Ca2+ and K+ currents, INaP was evoked by slow depolarizing voltage ramps or by long depolarizing voltage steps. The cholinergic agonist, carbachol, produced an atropine-sensitive decrease of INaP at all potentials. When applied at a saturating concentration (20 microM), carbachol reduced peak INaP by 38% on average. Carbachol did not alter the voltage dependence of INaP activation nor did it interfere with the slow inactivation of INaP. Our data indicate that INaP can be targeted by the rich cholinergic innervation of the neocortex. Because INaP is activated in the subthreshold voltage range, cholinergic inhibition of this current would be particularly suited to modulate the electrical behavior of neocortical pyramidal cells below and near firing threshold.

A special blocker reveals the presence and function of the hyperpolarization-activated cation current IH in peripheral mammalian nerve fibres.

Electrotonic responses recorded extra- or intracellularly from peripheral nerve preparations show a "sag" to hyperpolarizing current pulses. The biophysical nature of this "inward rectification" is still under discussion since the phenomenon has not been noted at voltage-clamped single nerve fibres, and since Cs+, which reduces inward rectification, is not a specific ion channel blocker. In this study, we found that low micromolar concentrations of ZD 7288, a specific blocker of the hyperpolarization-activated cationic current (Ih) in the soma of central mammalian neurons, result in a complete block of inward rectification in the electrotonic responses of isolated rat spinal dorsal roots. In addition, ZD 7288 enhanced the activity-dependent slowing of conduction seen in compound C fibre action potentials of isolated rat vagus nerves and augmented the post-tetanic hyperpolarization following trains of action potentials in unmyelinated and myelinated axons. The data suggest that ZD 7288 is a potent blocker and a useful research tool for the study of hyperpolarization-activated inward rectification (Ih) of peripheral nerve preparations.

Amplification of EPSPs by low Ni(2+)- and amiloride-sensitive Ca2+ channels in apical dendrites of rat CA1 pyramidal neurons.

Distal synaptic input to hippocampal CA1 pyramidal neurons was evoked by electrical stimulation of afferent fibers in outer stratum radiatum. Whole cell recordings from CA1 cell somata served to monitor excitatory postsynaptic potential (EPSP) envelopes after dendritic processing. To probe a functional role of low-voltage-activated Ca2+ current [or T current I(T)] in the apical dendrite, EPSP recordings were combined with local application of antagonists of I(T). Dendritic application of low concentrations of Ni2+ (5 microM) and amiloride (50 microM) reduced EPSP amplitude measured at the soma (resting membrane potential -70 mV) by 33.0 +/- 2.9% (mean +/- SE, n = 27) and 27.0 +/- 2.1% (n = 26), respectively. No appreciable effect on EPSP time course was observed. As expected from the voltage dependence of I(T) activation, the inhibitory effect of both antagonists was strongly attenuated when EPSPs were recorded at hyperpolarized membrane potential (-90 mV). In contrast to dendritic application, somatic application of Ni2+ or amiloride produced only weak reduction of EPSP amplitude. Our data indicate that dendritic low Ni(2+)- and amiloride-sensitive Ca2+ channels giving rise predominantly to I(T) can produce substantial amplification of synaptic input. We thus propose that these channels represent an important component of subthreshold signal integration in apical dendrites of CA1 pyramidal cells.

Function of the hyperpolarization-activated inward rectification in nonmyelinated peripheral rat and human axons.

The function of time-dependent, hyperpolarization-activated inward rectification was analyzed on compound potentials of nonmyelinated axons in the mammalian peripheral nervous system. Isolated rat vagus nerves and fascicles of biopsied human sural nerve were tested in a three-chambered, Vaseline-gap organ bath at 37 degrees C. Inward rectification was assessed by recording the effects of long-lasting hyperpolarizing currents on electrical excitability with the use of the method of threshold electrotonus (program QTRAC, copyright Institute of Neurology, London, UK) and by measuring activity-dependent changes in conduction velocity and membrane potential. Prominent time-dependent, cesium-sensitive inward rectification was revealed in rat vagus and human sural nerve by recording threshold electrotonus to 200-ms hyperpolarizing current pulses. A slowing of compound action potential conduction was observed during a gradual increase in the stimulation frequency from 0.1 to 3 Hz. Above a stimulation frequency of 0.3 Hz, this slowing of conduction was enhanced during bath application of 1 mM cesium. Cesium did not alter action potential waveforms during stimulation at frequencies < 1 Hz. Cesium-induced slowing in action potential conduction was correlated with membrane hyperpolarization. The hyperpolarization by cesium was stronger during higher stimulation frequencies and small in unstimulated nerves. These data show that a cesium-sensitive, time-dependent inward rectification in peripheral rat and human nonmyelinated nerve fibers limits the slowing in conduction seen in such axons at action potential frequencies higher than approximately 0.3 Hz.

Chronic clozapine versus chronic haloperidol treatment: differential effects on electrically evoked dopamine efflux in the rat caudate putamen, but not in the nucleus accumbens.

Fast cyclic voltammetry at carbon-fibre micro-electrodes was used to investigate the effects of chronic clozapine or haloperidol administration on electrically evoked dopamine efflux in the nucleus accumbens and caudate putamen of the anaesthetized rat. Stimulation trains were delivered to the median forebrain bundle (60 pulses, 350 microns duration) every 5 min, and the evoked dopamine efflux measured as a function of a) the applied stimulus intensity (range 0.2 mA-1.0 mA), and b) the applied stimulus frequency (range 10 Hz-250 Hz). Chronic administration of either clozapine (20 mg/kg x 21 days, p.o.) or haloperidol (1 mg/kg x 21 days, p.o.) significantly reduced electrically evoked dopamine efflux in the nucleus accumbens over the range of stimulus intensities and frequencies tested. The reduction in evoked dopamine efflux observed in the nucleus accumbens of clozapine- and haloperidol-treated rats showed no statistically significant difference. In contrast, only chronic haloperidol treatment significantly reduced evoked dopamine efflux in the caudate putamen. These findings demonstrate that chronic treatment with either the atypical neuroleptic, clozapine, or the typical neuroleptic, haloperidol, produce long-term changes in mesolimbic dopamine function; actions which may underlie their antipsychotic efficacy. They also provide further evidence that the sparing action of clozapine on nigrostriatal dopamine activity may underlie the lower incidence of extrapyramidal side effects associated with its long-term administration.

Dendritic Na+ channels amplify EPSPs in hippocampal CA1 pyramidal cells.

1. Whole cell recordings were performed on the somata of CA1 pyramidal neurons in the rat hippocampal slice preparation Remote synaptic events were evoked by electrical stimulation of Schaffer collateral/commissural fibers in outer stratum radiatum. To isolate non-N-methyl-D-aspartate (NMDA)-mediated excitatory postsynaptic potentials (EPSPs), bath solutions contained the NMDA receptor antagonist, D-2-amino-5-phosphonovaleric acid (D-APV; 30 microM), the gamma-aminobutyric acid-A (GABAA) receptor antagonist, bicuculline (10 microM), and the GABAB receptor antagonists, CGP 35348 (30 microM) or, in some experiments, saclofen (100 microM). 2. Local application of tetrodotoxin (TTX; 0.5-10 microM) into the proximal region of the apical dendrite reduced the peak amplitude of somatically recorded EPSPs by 28% on average. In contrast to dendritic TTX application, injection of TTX into the axosomatic region of the recorded neuron reduced EPSP amplitude by only 12% on average. 3. Spill-over of dendritically applied TTX into stratum pyramidale or into outer stratum radiatum was ruled out experimentally: somatic action potentials and field EPSPs recorded near the stimulation site in outer stratum radiatum remained unaffected by local TTX application. 4. Variations of somatic membrane potential revealed a strong voltage dependence of EPSP reduction after dendritic TTX application with the effect increasing substantially with membrane depolarization. Together with the field recordings from stratum radiatum, this finding argues strongly against a predominantly presynaptic site of TTX action. 5. We therefore ascribe the EPSP decrease after local TTX application to the proximal dendrite to suppression of dendritic Na+ channels, which we assume to give rise to a noninactivating (persistent) Na+ current (INaP) in the subthreshold voltage range. Our data suggest that presumed dendritic INaP produces considerable elevation of remote excitatory signals, thereby compensating for much of their electrotonic attenuation. 6. The experimental findings were related to computer simulations performed on a reduced compartmental model of the CA1 neuron. Because the experimental evidence available so far yields only indirect clues on the strength and distribution of INaP, we allowed considerable variations in these parameters. We also varied both size and location of synaptic input. 7. The major conclusions drawn from these simulations are the following: somatic INaP alone produces little EPSP enhancement; INaP density at the axon hillock/initial segment has to be at least twice the density at the soma to produce substantial EPSP amplification; depending on the density and distribution of dendritic INaP, < or = 80% of a remote synaptic potential arrives at the soma (compared with only 52% in a passive dendrite); synaptic potentials receive progressively more elevation by dendritic INaP the stronger they are; even if restricted to the proximal segment of the apical dendrite, INaP also affects dendritic processing at more distal segments; and spatial distribution rather than local density appears to be the most important parameter determining the role of dendritic INaP in synaptic integration.