Temporal offset between precipitation and water uptake of Mediterranean pine trees varies with elevation and season.

For climate models that use paleo-environment data to predict future climate change, tree-ring isotope variations are one important archive for the reconstruction of paleo-hydrological conditions. Due to the rather complicated pathway of water, starting from precipitation until its uptake by trees and the final incorporation of its components into tree-ring cellulose, a closer inspection of seasonal variations of tree water uptake is important. In this study, branch and needle samples of two pine species (Pinus pinaster and Pinus nigra subsp. laricio) and several water compartments (precipitation, creek, soil) were sampled over a two-year period and analyzed for the temporal variations of their oxygen and hydrogen stable isotope ratios (δ18O and δ2H) at five sites over an elevation gradient from sea level to around 1600 m a.s.l. on the Mediterranean island of Corsica (France). A new model was established to disentangle temporal relationships of source water uptake of trees. It uses a calculation method that incorporates the two processes mostly expected to affect source water composition: mixing of waters and evaporation. The model results showed that the temporal offset from precipitation to water uptake is not constant and varies with elevation and season. Overall, seasonal source water origin was shown to be dominated by precipitation from autumn and spring. While autumn precipitation was a more important water source for trees growing at mid- (~800-1000 m a.s.l) and high-elevation (~1600 m a.s.l.) sites, trees at coastal sites mostly took up water from late winter and spring. These findings show that predicted decreases in precipitation amounts during the wet season in the Mediterranean can have strong impacts on water availability for pine trees, especially at higher elevations.

Compartmental models of rat cerebellar Purkinje cells based on simultaneous somatic and dendritic patch-clamp recordings.

1. Simultaneous dendritic and somatic patch-clamp recordings were made from Purkinje cells in cerebellar slices from 12- to 21-day-old rats. Voltage responses to current impulses injected via either the dendritic or the somatic pipette were obtained in the presence of the selective I(h) blocker ZD 7288 and blockers of spontaneous synaptic input. Neurons were filled with biocytin for subsequent morphological reconstruction. 2. Four neurons were reconstructed and converted into detailed compartmental models. The specific membrane capacitance (C(m)), specific membrane resistance (R(m)) and intracellular resistivity (R(i)) were optimized by direct fitting of the model responses to the electrophysiological data from the same cell. Mean values were: C(m), 0.77 +/- 0.17 microF cm(-2) (mean +/- S.D.; range, 0.64-1.00 microF cm(-2)), R(m), 122 +/- 18 kOmega cm(2) (98-141 kOmega cm(2)) and R(i), 115 +/- 20 Omega cm (93-142 Omega cm). 3. The steady-state electrotonic architecture of these cells was compact under the experimental conditions used. However, somatic voltage-clamp recordings of parallel fibre and climbing fibre synaptic currents were substantially filtered and attenuated. 4. The detailed models were compared with a two-compartment model of Purkinje cells. The range of synaptic current kinetics that can be faithfully recorded using somatic voltage clamp is predicted fairly well by the two-compartment model, even though some of its underlying assumptions are violated. 5. A model of I(h) was constructed based on voltage-clamp data, and inserted into the passive compartmental models. Somatic EPSP amplitude was substantially attenuated compared to the amplitude of dendritic EPSPs at their site of generation. However, synaptic efficacy of the same quantal synaptic conductance, as measured by the somatic EPSP amplitude, was only weakly dependent on synaptic location on spiny branchlets. 6. The passive electrotonic structure of Purkinje cells is unusual in that the steady-state architecture is very compact, while voltage transients such as synaptic potentials and action potentials are heavily filtered.

Synaptic function: dendritic democracy.

Neurons receive synaptic inputs primarily onto their dendrites, which filter synaptic potentials as they spread toward the soma. Recent results indicate that this filtering appears to be compensated by increasing the synaptic conductance at distal synapses, thus normalizing the efficacy of synaptic inputs at the soma.

Propagation of action potentials in dendrites depends on dendritic morphology.

Action potential propagation links information processing in different regions of the dendritic tree. To examine the contribution of dendritic morphology to the efficacy of propagation, simulations were performed in detailed reconstructions of eight different neuronal types. With identical complements of voltage-gated channels, different dendritic morphologies exhibit distinct patterns of propagation. Remarkably, the range of backpropagation efficacies observed experimentally can be reproduced by the variations in dendritic morphology alone. Dendritic geometry also determines the extent to which modulation of channel densities can affect propagation. Thus in Purkinje cells and dopamine neurons, backpropagation is relatively insensitive to changes in channel densities, whereas in pyramidal cells, backpropagation can be modulated over a wide range. We also demonstrate that forward propagation of dendritically initiated action potentials is influenced by morphology in a similar manner. We show that these functional consequences of the differences in dendritic geometries can be explained quantitatively using simple anatomical measures of dendritic branching patterns, which are captured in a reduced model of dendritic geometry. These findings indicate that differences in dendritic geometry act in concert with differences in voltage-gated channel density and kinetics to generate the diversity in dendritic action potential propagation observed between neurons. They also suggest that changes in dendritic geometry during development and plasticity will critically affect propagation. By determining the spatial pattern of action potential signaling, dendritic morphology thus helps to define the size and interdependence of functional compartments in the neuron.

Dendritic coincidence detection of EPSPs and action potentials.

We describe a mechanism for coincidence detection mediated by the interaction between backpropagating action potentials and EPSPs in neocortical pyramidal neurons. At distal dendritic locations, appropriately timed EPSPs or oscillations could increase the amplitude of backpropagating action potentials by three- to fourfold. This amplification was greatest when action potentials occurred at the peak of EPSPs or dendritic oscillations and could lead to somatic burst firing. The increase in amplitude required sodium channel activation but not potassium channel inactivation. The temporal characteristics of this amplification are similar to those required for changes in synaptic strength, suggesting that this mechanism may be involved in the induction of synaptic plasticity.

Differential shunting of EPSPs by action potentials.

Neurons encode information and communicate via action potentials, which are generated following the summation of synaptic events. It is commonly assumed that action potentials reset the membrane potential completely, allowing another round of synaptic integration to begin. We show here that the conductances underlying the action potential act instead as a variable reset of synaptic integration. The strength of this reset is cell type-specific and depends on the kinetics, location, and timing of the synaptic input. As a consequence, distal synapses, as well as inputs mediated by N-methyl-d-aspartate receptor activation, can contribute disproportionately to synaptic integration during action potential firing.

Coincidence detection in single dendritic spines mediated by calcium release.

Cerebellar long-term depression (LTD) is a calcium-dependent process in which coincident activity of parallel fiber (PF) and climbing fiber (CF) synapses causes a long-lasting decrease in PF synaptic strength onto Purkinje cells. Here we show that pairing CF activation with bursts of PF activity triggers large (>10 microM) calcium signals in Purkinje cell dendrites. When PFs are densely activated, signals span whole dendritic branchlets and are mediated by voltage-dependent calcium entry. When PFs are sparsely activated, however, signals are restricted to single spines and blocked by metabotropic glutamate receptor antagonists. Single-spine signals and sparse-stimulation LTD are also blocked by thapsigargin, indicating that calcium must be released from stores. Single-spine signals and sparse-stimulation LTD are greatest when PF activation precedes the CF activation within 50-200 ms. This timing rule matches the properties of several forms of motor learning, providing a link between behavior and functional properties of cerebellar synaptic plasticity.

Diversity and dynamics of dendritic signaling.

Communication between neurons in the brain occurs primarily through synapses made onto elaborate treelike structures called dendrites. New electrical and optical recording techniques have led to tremendous advances in our understanding of how dendrites contribute to neuronal computation in the mammalian brain. The varied morphology and electrical and chemical properties of dendrites enable a spectrum of local and long-range signaling, defining the input-output relationship of neurons and the rules for induction of synaptic plasticity. In this way, diversity in dendritic signaling allows individual neurons to carry out specialized functions within their respective networks.

The core-shell dichotomy of nucleus accumbens in the rhesus monkey as revealed by double-immunofluorescence and morphology of cholinergic interneurons.

Double-immunolabelling experiments for the combinations, calretinin (CR)-calbindin, CR-tyrosine hydroxylase (TH) and calbindin-TH, were performed in rhesus monkeys to compare the chemical organization of the nucleus accumbens (ACC) in primates and rodents. Additionally, the soma sizes and numbers of primary dendrites of cholinergic neurons in the subregions of ACC were compared with those of caudate-putamen. Our findings subserve the shell-core concept also in the primate ACC, as like in the rat, CR immunoreactivity (-ir) due to intense neuropil labelling is very strong in the shell of rhesus monkey, but poor in the core. The staining intensity of this marker decreases in dorsoventral direction. An almost complementary pattern was noted in sections of the monkey ACC immunostained for both calbindin and TH. The cholinergic interneurons of the nucleus caudatus-putamen are clearly distinguished from those of the ACC and insula Calleja magna by their much bigger soma sizes and higher numbers of primary dendrites. Cholinergic neurons of the shell were found to be slightly, but significantly, larger than those of the core that also subserves subdivision of the primate ACC into shell and core. A low proportion of tyrosine-hydroxylase-immunostained cells, already previously described below the rostral ACC, co-expressed CR but not calbindin. A CR-immunoreactive neuronal population, intermingled with these cells, extends as a stripe medially to the ACC along the septal part of corpus callosum into the lateral septal area. The presumed origin of CR-immunoreactive fibres in the shell of ACC is discussed.

Estimating the time course of the excitatory synaptic conductance in neocortical pyramidal cells using a novel voltage jump method.

We introduce a method that permits faithful extraction of the decay time course of the synaptic conductance independent of dendritic geometry and the electrotonic location of the synapse. The method is based on the experimental procedure of Pearce (1993), consisting of a series of identical somatic voltage jumps repeated at various times relative to the onset of the synaptic conductance. The progression of synaptic charge recovered by successive jumps has a characteristic shape, which can be described by an analytical function consisting of sums of exponentials. The voltage jump method was tested with simulations using simple equivalent cylinder cable models as well as detailed compartmental models of pyramidal cells. The decay time course of the synaptic conductance could be estimated with high accuracy, even with high series resistances, low membrane resistances, and electrotonically remote, distributed synapses. The method also provides the time course of the voltage change at the synapse in response to a somatic voltage-clamp step and thus may be useful for constraining compartmental models and estimating the relative electrotonic distance of synapses. In conjunction with an estimate of the attenuation of synaptic charge, the method also permits recovery of the amplitude of the synaptic conductance. We use the method experimentally to determine the decay time course of excitatory synaptic conductances in neocortical pyramidal cells. The relatively rapid decay time constant we have estimated (tau approximately 1.7 msec at 35 degrees C) has important consequences for dendritic integration of synaptic input by these neurons.

Tonic synaptic inhibition modulates neuronal output pattern and spatiotemporal synaptic integration.

Irregular firing patterns are observed in most central neurons in vivo, but their origin is controversial. Here, we show that two types of inhibitory neurons in the cerebellar cortex fire spontaneously and regularly in the absence of synaptic input but generate an irregular firing pattern in the presence of tonic synaptic inhibition. Paired recordings between synaptically connected neurons revealed that single action potentials in inhibitory interneurons cause highly variable delays in action potential firing in their postsynaptic cells. Activity in single and multiple inhibitory interneurons also significantly reduces postsynaptic membrane time constant and input resistance. These findings suggest that the time window for synaptic integration is a dynamic variable modulated by the level of tonic inhibition, and that rate coding and temporal coding strategies may be used in parallel in the same cell type.

Intersynaptic diffusion of neurotransmitter.

According to many theories of brain function, the computational power of the brain depends upon the number of independent synapses it contains. A synapse will not be independent if its receptors are activated or modified by neurotransmitter released at neighbouring synapses. Recently, there have been several reports suggesting the occurrence of 'crosstalk' or 'spillover', and a large number of results consistent with crosstalk. However, the quantitative importance of this phenomenon remains uncertain. We estimate the significance of crosstalk using a simple model which predicts that, during concentrated synaptic activity, crosstalk between distinct synapses is likely to activate high-affinity receptors and may also desensitize certain receptors. Comparison of these predictions with the experimental data highlights the information that is required for a more detailed model of crosstalk.

Dendritic and somatic glutamate receptor channels in rat cerebellar Purkinje cells.

1. The properties of glutamate receptor (GluR) channels in outside-out patches from the dendrites and somata of rat cerebellar Purkinje cells in brain slice were studied using fast agonist application techniques. Dendritic patches were isolated 40-130 micronm from the soma. 2. Outside-out patches from both dendrites and somata of Purkinje cells responded to application of glutamate with a current which desensitized rapidly and nearly completely. Currents evoked by glutamate application were blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), were mimicked by L-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), and were modulated by cyclothiazide. Kainate produced small, non-desensitizing currents. No currents were observed in response to aspartate application. Responses characteristic of NMDA receptor activation were not observed. These findings indicate that glutamate-activated currents were mediated by the AMPA subtype of GluR. 3. Deactivation of the GluR channels following 1 ms pulses of glutamate occurred with a time constant of 1.23 +/- 0.07 ms in dendritic and 1.12 +/- 0.04 ms in somatic patches. Desensitization occurred with a time constant of 5.37 +/- 0.26 ms in dendritic and 5.29 +/- 0.29 ms in somatic patches. The time constant of recovery from desensitization caused by a 1 ms application of 1 mM glutamate was 36 ms in dendritic patches and 33 ms in somatic patches. 4. Half-maximal activation of the GluR channels was achieved at a glutamate concentration of 432 microM. Deactivation kinetics were not dependent on the glutamate concentration, while desensitization became slower at lower glutamate concentrations. 5. Pre-equilibration of patches with low concentrations of glutamate reduced the peak current activated by 1 mM glutamate. The IC50 for this effect was 8.7 microM. Equilibrium desensitization did not affect the kinetics of the current activated by 1 mM glutamate. 6. The current-voltage relationship of the peak current was linear in normal Na(+)-rich external solution, with a reversal potential near 0 mV. In Ca(2+) -rich external solution, the reversal potentials were -51.4 +/- 2.9 and -51.5 +/- 2.8 mV for dendritic and somatic patches, respectively, indicating that these glutamate channels have a low permeability to Ca2+ (PCa/PCa = 0.053). 7. The mean single-channel conductance of the GluR channels measured using non-stationary fluctuation analysis was approximately 8 pS in dendritic and somatic patches, and the maximum open probability was at least 0.7 with 5 mM glutamate. 8. GluR channel kinetics in patches excised from the soma of neonatal (postnatal day 4; P4) Purkinje cells, before the development of the dendritic arborization of the Purkinje cell, were similar to those in patches excised from more mature (P12-18) Purkinje cells. 9. Dendritic and somatic GluR channels in Purkinje cells appear to be functionally identical, are AMPA-subtype receptors containing the GluR-B subunit, and have rapid kinetics and low permeability to Ca2+. A kinetic model was constructed which faithfully reproduces the gating characteristics of the GluR channels.

Action potential initiation and backpropagation in neurons of the mammalian CNS.

Most neurons in the mammalian CNS encode and transmit information via action potentials. Knowledge of where these electrical events are initiated and how they propagate within neurons is therefore fundamental to an understanding of neuronal function. While work from the 1950s suggested that action potentials are initiated in the axon, many subsequent investigations have suggested that action potentials can also be initiated in the dendrites. Recently, experiments using simultaneous patch-pipette recordings from different locations on the same neuron have been used to address this issue directly. These studies show that the site of action potential initiation is in the axon, even when synaptic activation is powerful enough to elicit dendritic electrogenesis. Furthermore, these and other studies also show that following initiation, action potentials actively backpropagate into the dendrites of many neuronal types, providing a retrograde signal of neuronal output to the dendritic tree.

Axonal initiation and active dendritic propagation of action potentials in substantia nigra neurons.

The site of action potential initiation in substantia nigra neurons was investigated by using simultaneous somatic and dendritic whole-cell recording in brain slices. In many dopamine neurons, action potentials were observed first at the dendritic recording site. Anatomical reconstruction showed that in these neurons, the axon emerged from the dendrite from which the recording had been made. Action potentials showed little attention in the dendritic tree, which in dopamine neurons was shown to be due to recruitment of dendritic sodium channels and may be related to the dendritic release of dopamine. We conclude that in substantia nigra neurons, the site of action potential initiation, and thus the final site of synaptic integration, is in the axon. As the axon can originate from a dendrite, up to 240 microns away from the soma, synaptic input to the axon-bearing dendrite may be privileged with respect to its ability to influence action potential initiation.

Patch test results with serial dilutions of nickel sulfate (with and without detergent), palladium chloride, and nickel and palladium metal plates.

Clinical experience suggests the existence of different degrees of sensitivity in nickel-allergic patients. For quantification of this phenomenon, 462 consecutive patients with previously diagnosed or strongly suspected nickel allergy were tested with serial dilution patch tests with 5 ppm to 5% nickel sulfate in pet. (Ni), and 5 ppm to 1% nickel sulfate in pet. with 1% detergent (Ni/D). Additionally, nickel and palladium metal plates were tested in 103, and cobalt salts, dichromate and palladium chloride (PdCl2) in most patients. 332 patients reacted positively to Ni or Ni/D. The influence of a concomitantly administered detergent was not significant. A significant correlation was found between positive reactions to low concentrations of Ni (or Ni/D), i.e., 0.1% or less (N = 166), and concomitant reactions to nickel metal plates, cobalt salts and PdCl2 and a history of ear piercing with metal intolerance. The clinical relevance of reactions to PdCl2 is at present not clear. A subgroup of nickel-allergic patients with "high sensitivity" can be defined. In future studies further addressing the clinical relevance of high versus low sensitivity, patch testing with 0.01, 0.1, 1.0 and 5% nickel sulfate in pet. is recommended instead of routine tests with 5% only.

Inhibitory synaptic potentials in guinea-pig substantia nigra dopamine neurones in vitro.

1. The properties of stimulus-evoked and spontaneous inhibitory synaptic potentials were examined in guinea-pig substantia nigra dopamine neurones in sagittal and coronal midbrain slices in the presence of glutamate receptor antagonists. 2. Focal electrical stimulation within the substantia nigra, cerebral peduncle, internal capsule or the striatum evoked a biphasic IPSP consisting of a fast and a slow component, with peak latencies of about 30 and 250 ms, respectively. The fast component was sensitive to chloride injection, reversed polarity at -79.4 +/- 1.1 mV and was blocked by the GABAA receptor antagonists picrotoxin and bicuculline. The slow IPSP reversed at -99.3 +/- 5.4 mV and was blocked by the GABAB receptor antagonists 2-hydroxysaclofen and CGP 35348. 3. Spontaneous IPSPs were observed in many neurones. These events reversed polarity at -77.5 +/- 2.6 mV and were completely blocked by bicuculline and/or picrotoxin. In the presence of TTX, small spontaneous events remained which probably represent miniature IPSPs. In coronal slices, application of 4-aminopyridine raised the frequency of spontaneous IPSPs, presumably by activating nigral interneurones, but failed to reveal spontaneous biphasic IPSPs or spontaneous pure slow IPSPs. 4. The amplitude of the fast IPSPs fluctuated from trial to trial. Amplitude histograms of minimal fast IPSPs displayed evenly spaced peaks, suggesting that synaptic transmission is quantal at these synapses. The measured peak spacing depended on the driving force for Cl-. 5. The fast IPSP showed little or no paired-pulse depression, and in the presence of 2-hydroxysaclofen (400-600 microM) showed paired-pulse facilitation. The GABAB agonist baclofen inhibited the fast IPSP via a presynaptic mechanism. The pharmacologically isolated slow IPSP showed marked paired-pulse facilitation. 6. It is concluded that synaptic inhibition in the substantia nigra is mediated by GABA, is relatively resistant to frequency-dependent depression and is regulated by presynaptic GABAB autoreceptors. Striatonigral and pallidonigral fibres activate both GABAA and GABAB receptors, while intranigral pathways appear to activate predominantly GABAA receptors.