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Modeling extracellular field potentials and the frequency-filtering properties of extracellular space.

Extracellular local field potentials are usually modeled as arising from a set of current sources embedded in a homogeneous extracellular medium. Although this formalism can successfully model several properties of extracellular local field potentials, it does not account for their frequency-dependent attenuation with distance, a property essential to correctly model extracellular spikes. Here we derive expressions for the extracellular potential that include this frequency-dependent attenuation. We first show that, if the extracellular conductivity is nonhomogeneous, there is induction of nonhomogeneous charge densities that may result in a low-pass filter. We next derive a simplified model consisting of a punctual (or spherical) current source with spherically symmetric conductivity/permittivity gradients around the source. We analyze the effect of different radial profiles of conductivity and permittivity on the frequency-filtering behavior of this model. We show that this simple model generally displays low-pass filtering behavior, in which fast electrical events (such as Na(+)-mediated action potentials) attenuate very steeply with distance, whereas slower (K(+)-mediated) events propagate over larger distances in extracellular space, in qualitative agreement with experimental observations. This simple model can be used to obtain frequency-dependent extracellular field potentials without taking into account explicitly the complex folding of extracellular space.

Action Potentials↗

Stimulus-induced changes in extracellular Na+ and Cl- concentration in relation to changes in the size of the extracellular space.

Extracellular Na+ - and Cl- -concentrations ([Na+]o, [Cl-]o) were recorded with ion-selective microelectrodes during repetitive stimulation and stimulus-induced self-sustained neuronal afterdischarges (SAD) in the sensorimotor cortex of cats. In all cortical layers [Na+]o initially decreased by 4-7 mM. In depths of more than 600 micrometer below the cortical surface such decreases usually turned into increases of 2-6 mM during the course of the SADs, whereas in superficial layers [Na+]o never rose above its resting level. [Cl-]o always showed an increase in the course of the SADs often preceded by an initial small decrease. The average increase at a depth of 1,000 micrometers was about 7 mM. [Cl-]o reached peak values at about the end of the ictal period, whereas [Na+]o reached its maximum shortly after the end of the SAD, at times when [K+]o was still elevated above the baseline concentration. These data indicate that the extracellular osmolarity can increase during SAD by up to 30 mM. Such an increase in osmolarity can be explained by an increase in the number of intracellular particles, caused by cleavage of larger molecules during enhanced metabolism. This could lead to cell-swelling due to passive water influx from the extracellular space (ES). However, the resulting reduction of the size of the ES is calculated to be less than 10% for an increase in intracellular osmolarity by 30 mOsm. This value is too small as compared to previously measured ES-reductions under similar conditions (i.e., 30% reduction at 1,000 micrometers; Dietzel et al. 1980). Reductions of the size of the ES that accompany the observed changes in the ionic environment, are quantitatively explained on the basis of the extended glial buffering mechanism described in the preceding paper.

Animals↗

Contribution of shrinkage of extracellular space to extracellular K+ accumulation in myocardial ischaemia of the rabbit.

1. The contribution of the concentrating effect due to shrinkage of the extracellular space (ECS) to cellular K+ efflux on extracellular potassium ([K+]o) accumulation in response to ischaemia was investigated in an isolated, blood-perfused rabbit papillary muscle preparation with a confined extracellular space. 2. The ECS was quantified using either of two extracellular markers, choline or tetramethyl ammonium (TMA), each with specific ion-selective electrodes, as well as by measurement of extracellular resistance (ro). [K+]o and [Na+]o were also measured simultaneously using K(+)- and Na(+)-selective electrodes. 3. During ischaemia, [K+]o increased 3-fold from 4.2 +/- 0.1 to 12.6 +/- 1.0 mM at 10 min (n = 10) analogous to changes in the ischaemic heart in vivo. The ECS decreased to 83.9 +/- 3.2% of control measured using 1 mM choline extracellularly (n = 9, P < 0.01) or to 85.7 +/- 0.7% of control using 1 mM TMA (n = 6, P < 0.01). Nearly identical decreases in ro (84.1 +/- 2.4%, n = 15, P < 0.01) occurred simultaneously. 4. The small decrease in the ECS contributed only 0.8-0.9 mM to the total increase in [K+]o of 8.4 mM and had a minor effect on transmembrane K+ flux. No significant differences between the relative changes in [choline] and [Na+]o were observed. This excluded a major transmembrane Na+ movement during early ischaemia. 5. Bumetanide (10 mM), an inhibitor of K(+)-Cl- cotransport, a process which is involved in cell volume regulation consequent to osmotic cell swelling, significantly attenuated the increase in [K+]o after 6 min of ischaemia (8.3 +/- 0.6 mM, n = 5 vs. 10.3 +/- 0.4 mM in the control group, n = 6, P < 0.05), whereas N-ethylmaleimide (1 mM), a stimulator of this cotransporter, augmented [K+]o accumulation (12.0 +/- 0.6 mM at 6 min, P < 0.05). 6. We conclude that during early myocardial ischaemia, a major component of [K+]o accumulation is not caused by diminution of ECS per se, but rather by increased net K+ efflux due in part to K(+)-Cl cotransport secondary to myocyte volume regulation.

Action Potentials↗

Temporal profile of changes in brain tissue extracellular space and extracellular ion (Na(+), K(+)) concentrations after cerebral ischemia and the effects of mild cerebral hypothermia.

Cerebral ischemic cellular swelling occurs primarily in astrocytes. This water influx into the intracellular space is believed to result from osmotic water movement after disruption of membrane ionic homeostasis. However, cellular swelling occurs earlier than expected after ischemia and new ionic and water channels have been discovered. This study examined the temporal profile of the water and ionic movement across the cell membrane after global ischemia by measuring the changes in extracellular space (ECS), extracellular K(+) and Na(+) ion concentrations ([K(+)](e) and [Na(+)](e)) using a high resolution tissue impedance probe and ion selective micropipettes in the rat cortex. The effect of mild cerebral hypothermia (31.5 +/- 2.6 degrees C brain temperature) on these parameters was also examined. The ECS started to decrease at 34 +/- 8 sec after global ischemia and reached half the maximum change at 61 +/- 17 sec. [K(+)](e) started to increase initially at 33 +/- 11 sec (phase 1) and then increased rapidly at 62 +/- 25 sec (phase 2), and [Na(+)](e) started to decrease at 88 +/- 27 sec after ischemia. With mild hypothermia, the ECS started to decrease at 75 +/- 35 sec after ischemia and reached half the maximum change at 123 +/- 44 sec, [K(+)](e) started to increase initially at 80 +/- 24 sec (phase 1) and then increased rapidly at 120 +/- 32 sec (phase 2), and [Na(+)](e) started to decrease at 172 +/- 70 sec. The present study shows that ischemic cellular swelling (decreased ECS) occurs concomitantly with the phase 1 increase of [K(+)](e) but precedes the disruption of ionic membrane homeostasis (phase 2). Mild hypothermia prolongs the onset of these phenomena but does not affect the magnitude of the changes in ECS and ion concentrations.

Animals↗

Estimation of extracellular space and blood volume using bioelectrical impedance measurements.

The bromide-82 dilution space (extracellular space, ECS) and blood volume (BV) were measured in 21 patients with esophageal and gastric cancer and in 27 patients 18-96 months after total gastrectomy. Resistance (R) and reactance (Xc) from bioelectrical impedance measurements were used to obtain multiple regression equations for ECS and BV. The variables weight, gender, and height 2/Xc were independent predictors of ECS (r = 0.767; P less than 0.0001). Height 2/R and gender were predictors of blood volume (r = 0.856; P less than 0.0001). The mean difference between the Br space and the ECS predicted from impedance measurements was 0 +/- 1.54 (mean +/- SD). The limits of agreement (+/- 2 SD) were therefore +/- 3.08 l or 19.6% of the mean Br space of 15.7 l. The limits of agreement for BV were +/- 789 ml or +/- 19.7% of the average BV of 4008 ml. It is concluded that bioelectrical impedance plethysmography using a single frequency can be used for the estimation of ECS and BV. The wide limits of agreement, however, may limit its used in clinical practice.

Adult↗

Cell cavities increase tortuosity in brain extracellular space.

Brain extracellular space (ECS) forms hindered pathways for molecular diffusion in chemical signaling and drug delivery. Hindrance is quantified by the tortuosity lambda; the tortuosity obtained from simulations using uniformly spaced convex cells is significantly lower than that measured experimentally. To attempt to account for the difference in results, this study employed a variety of ECS models based on an array of cubic cells containing open rectangular cavities that provided the ECS with dead-space microdomains. Monte Carlo simulations demonstrated that, in such ECS models, lambda can equal or exceed the typical experimental value of about 1.6. The simulations further revealed that lambda is relatively independent of cavity shape and the number of cavities per cell. It mainly depends on the total ECS volume fraction alpha, the cavity volume fraction alpha(c), and whether the cavity is located at the center of a cell face or formed at the junction of multiple cells. To describe the results from the different ECS models, an expression was obtained that related lambda to alpha, alpha(c), and an empirical exit factor beta that correlated with the ease with which a molecule could leave a cavity and its vicinity.

Biological Transport↗

Changes in diffusion through the brain extracellular space.

ECS (extracellular space) works as the microenvironment of brain cells. Diffusion through ECS may be described through an effective diffusion coefficient, D (e), which in turn depends on ECS porosity, epsilon, and tortuosity, T. In the present research, diffusion data together with epsilon and T were collected from the specialized literature and analysed to seek a correlation of T versus epsilon. On the basis of D (e) data, upper and lower T boundaries were defined and related to topologically 'dense' and 'loose' cell arrangement. A possible range for T variation was obtained for ECS, with epsilon ranging from 0.05 to 0.6. A tortuosity index ( n ) in the form of T and epsilon logarithmic ratio was introduced. This index may be adopted for recalculation of T or epsilon if only one of these parameters is known. As a result of data analysis and modelling, it was concluded that, upon different external conditions, for instance oxygen depletion, the ECS porosity decreases and cells (presumably through membrane rearrangements) adjust the void space to keep the diffusion within a defined range, which gives the living tissue the ability to maintain the diffusion level up to two or more times higher than in conventional granular bed packing. Thus, even with a dramatic ECS decrease, the cellular system is still able to support a given diffusion by decreasing the value of T. The obtained results clearly show the existence of three data clusters: a region of normal brain functioning, both for young and adult brains, for values of epsilon comprised between 0.15 and 0.30, and two regions of abnormal brain behaviour to the left and to the right of the normal region, corresponding to different states (aging, tumours, anoxia, brain death, etc.). The present approach allows defining the optimal range of epsilon and T to assure the best ECS diffusion efficiency for a specified macromolecule. This might be important in brain clinical treatment.

Animals↗

Contribution of dead-space microdomains to tortuosity of brain extracellular space.

The extracellular space (ECS) of the brain is a major channel for intercellular communication, nutrient and metabolite trafficking, and drug delivery. The dominant transport mechanism is diffusion, which is governed by two structural parameters, tortuosity and volume fraction. Tortuosity (lambda) represents the hindrance imposed on the diffusing molecules by the tissue in comparison with an obstacle-free medium, while volume fraction (alpha) is the proportion of tissue volume occupied by the ECS. Diffusion of small ECS markers can be exploited to measure lambda and alpha. In healthy brain tissue, lambda is about 1.6 but increases to 1.9-2.0 in pathologies that involve cellular swelling. Previously it was thought that lambda could be explained by the circumnavigation of diffusing molecules around cells. Numerical models of assemblies of convex cells, however, give an upper limit of about 1.23 for lambda. Therefore, additional factors must be responsible for lambda in brain. In principle, two mechanisms could account for the measured value: a more complex ECS geometry or an extracellular macromolecular matrix. Here we review recent work in ischemic tissue suggesting concave geometrical formations, dead-space microdomains, as a major determinant of extracellular tortuosity. A theoretical model of lambda based on diffusion dwell times supports this hypothesis and predicts that, in ischemia, dead spaces occupy approximately 60% of ECS volume fraction leaving only approximately 40% for well-connected channels. It is further proposed that dead spaces are present in healthy brain tissue where they constitute about 40% of alpha. The presence of dead-space microdomains in the ECS implies microscopic heterogeneity of extracellular channels with fundamental implications for molecular transport in brain.

Animals↗

Intrinsic optical signals in vitro: a tool to measure alterations in extracellular space with two-dimensional resolution.

In excitable tissues, extensive neuronal activity or pathophysiological conditions, such as spreading depression, ischemic infarct, or epileptic seizure, are accompanied by changes in extracellular space volume. Extracellular space volume, in turn, influences neuronal excitability and extracellular ion concentrations and is, therefore, an important parameter of brain activity. Unfortunately, determination of changes in extracellular space by ion-selective microelectrodes is tedious, restricted to one spot in space at a time and limited in time resolution. In this study we present intrinsic optical signals in vitro as a tool to measure relative changes in extracellular space volume in brain slice preparations with two-dimensional spatial and sufficient time resolution. Evidence is given that the intensity of intrinsic optical signals is linearly correlated to the amplitude of extracellular space volume changes. In contrast, the optical signal is poorly correlated to the concomitant increase in extracellular potassium concentration. We conclude that intrinsic optical signals in vitro are a useful tool to measure the spread of changes in extracellular space volume with high resolution in time and space. In combination with the measurement of the extracellular space at one location using ion-selective microelectrodes, it is possible to calibrate the optical signal to percentile alterations of extracellular space volume.

Animals↗

Influence of fixative osmolality on the morphometric determination of extracellular space in normal and reperfused ischaemic myocardium.

Morphometric determination of extracellular space in control and post-ischaemic reperfused rabbit myocardium was evaluated using two fixatives differing in their composition and total osmolality. Measurement of control extracellular space in an isotonic fixative (294 mOsm/kg water) was 20.8% and in a hypertonic fixative (1816 mOsm/kg water) was 22.2%. These values were not statistically different. Ischaemic durations of 15, 30, 60 and 90 min, followed by an equivalent period of reperfusion, created significant increases in extracellular space. The size of the extracellular space determined by both fixatives was found to be the same. Total fixative osmolality does not appear to influence morphometric evaluation of the extracellular space in control tissue or in tissue damaged by ischaemia and reperfusion.

Animals↗

Frequency and time domain studies of the micro-EEG from the brain extracellular space.

In these studies, recordings from the brain extracellular space of cats are considered. Macro and microelectrodes are utilized. Special studies using visual, auditory stimuli and certain drugs are done. Coherence, power phase and partial coherence spectra of wave field potentials and unit potentials are investigated. The electrographic profile of the micro electroencephalogram (EEG) and micro evoked response (ER) of the extracellular space is different than the macropotentials of the macro (EEG) and the macro (ER). The electrographic patterns of the micro (EEG) and the micro (ER) will vary with the microelectrode size, location and the variable microanatomy. There are many single neuron field potentials differentiated in space and time. In these unit fields there are varying electronic interactions varying in space and time. There are patterned inputs varying in space and time. There are fluctuating microfields distributed in overlapping layers, columns, slabs and patches and periaxonal extracellular spaces of the cerebral and cerebellar white matter. In these anatomical situations there are variable, degrees of local and general selective patterns of synchronization with variable direction, and selective spread. The extracellular space micro (EEG) shows complex and dynamic shifts of the micro (EEG) to the macro (EEG) frequencies and tuned macrofields.

Animals↗

Issues involved in the transmission of chemical signals through the brain extracellular space.

Two classes of substances exist within the extracellular space: energetic and informational. Examples of the former are glucose, dissolved oxygen and CO2 while the latter include excitatory amino acids, cathecholamines and opiates. The simple ions Na+ and Cl- are generally associated with energetic processes while extracellular K+ and Ca2+ tend to be informational in function. Local release of an informational substance brings about a concentration gradient that causes the substance to be dispersed in the extracellular space by diffusion. This process is modified relative to a free aqueous medium by the constraints of volume fraction, tortuosity and uptake. Volume fraction is defined simply as the fraction of a brain region that is extracellular. If a given quantity of substance is released into a region with a reduced volume fraction then the substance will reach a higher concentration than it would in a free medium. Tortuosity is related to the increase in the path length of the random walk of a diffusing particle due to the necessity to navigate around cellular obstructions. Tortuosity manifests itself as a decrease in the diffusion coefficient. Uptake represents the movement of a substance from the extracellular space to the intracellular. Since initially a concentration gradient exists in this direction and all membranes have some permeability some concentration-dependent uptake always occurs. In addition there exist specific carrier-mediated uptake processes for some substances such as amino acids or catecholamines. In some regions the dispersal process can be dominated by uptake rather than diffusion. While volume fraction, tortuosity and uptake have all been demonstrated by a technique based on the use of radiolabels and other methods, these classical techniques have limited spatial and temporal resolution. The advent of methods based on micro-injection of substances by iontophoresis or pressure and subsequent detection with ion-selective microelectrodes (ISMs) or voltammetric microsensors (VMs) has opened a new window onto the dynamic local behavior of the extracellular space. In the last decade our laboratory and others have studied the migration of the test substances tetramethylammonium, tetraethylammonium, AsF6- and alpha naphthalene sulfonate, the endogenous ions K+ and Ca2+, the epileptogenic agent penicillin and the neurotransmitter dopamine. These studies have been carried out on the cerebellum and some other regions in a variety of species that include rat, turtle, skate and an intervertebrate, the cuttlefish.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Maximum geometrical hindrance to diffusion in brain extracellular space surrounding uniformly spaced convex cells.

Brain extracellular space (ECS) constitutes a porous medium in which diffusion is subject to hindrance, described by tortuosity, lambda = (D/D*)1/2, where D is the free diffusion coefficient and D* is the effective diffusion coefficient in brain. Experiments show that lambda is typically 1.6 in normal brain tissue although variations occur in specialized brain regions. In contrast, different theoretical models of cellular assemblies give ambiguous results: they either predict lambda-values similar to experimental data or indicate values of about 1.2. Here we constructed three different ECS geometries involving tens of thousands of cells and performed Monte Carlo simulation of 3-D diffusion. We conclude that the geometrical hindrance in the ECS surrounding uniformly spaced convex cells is independent of the cell shape and only depends on the volume fraction alpha (the ratio of the ECS volume to the whole tissue volume). This dependence can be described by the relation lambda = ((3-alpha)/2)1/2, indicating that the geometrical hindrance in such ECS cannot account for lambda > 1.225. Reasons for the discrepancy between the theoretical and experimental tortuosity values are discussed.

Brain↗

In vivo diffusion analysis with quantum dots and dextrans predicts the width of brain extracellular space.

Diffusion within the extracellular space (ECS) of the brain is necessary for chemical signaling and for neurons and glia to access nutrients and therapeutics; however, the width of the ECS in living tissue remains unknown. We used integrative optical imaging to show that dextrans and water-soluble quantum dots with Stokes-Einstein diameters as large as 35 nm diffuse within the ECS of adult rat neocortex in vivo. Modeling the ECS as fluid-filled "pores" predicts a normal width of 38-64 nm, at least 2-fold greater than estimates from EM of fixed tissue. ECS width falls below 10 nm after terminal ischemia, a likely explanation for the small ECS visualized in electron micrographs. Our results will improve modeling of neurotransmitter spread after spillover and ectopic release and establish size limits for diffusion of drug delivery vectors such as viruses, liposomes, and nanoparticles in brain ECS.

Animals↗

Comparison of extracellular space in the mature and aging rat brain using a new technique.

A new technique for measuring extracellular space in the rat brain has been developed. It involves opening the blood-brain barrier with a bolus of hyperosmotic sucrose followed by a high-pressure perfusion of the cerebral vasculature with an isotonic solution containing an impermeant radioactive tracer, [3H]sucrose. After allowing the concentration of tracer in the brain to reach a plateau, the amount of radioactivity/mg of brain tissue is expressed as a percentage of the amount of radioactivity/mg of perfusate to obtain a value for extracellular space. The addition of glutaraldehyde to the perfusate results in the brain being fixed simultaneously for electron microscopy. Reproducible estimates of extracellular space were obtained similar to those obtained by other methods (e.g. Levin et al. 1970). As it is impossible to be sure of the validity of the absolute value of extracellular space obtained by any method using perfused solutions we have used our method for comparative purposes. Extracellular space was measured in mature (control) and ageing rats to test the claim that the volume of space in the cerebral cortex is substantially reduced with ageing (Bondareff and Narotsky 1972). We found a consistent tendency for the extracellular space to increase with age in the 6 regions of brain examined. This was not statistically significant except in the group of ageing rats on a food-restricted diet. Therefore, these results do not support a generalisation that the extracellular space decreases in the ageing brain. In both control and ageing rats, extracellular space was shown to be unevenly distributed in the brain, the largest space being present in the cerebellum, olfactory bulb, inferior and superior colliculi and the least space in the white matter. These are the first measurements of extracellular space in which assessment is possible by both electron microscopy and by the measurement of a chemical tracer.

Aging↗

Blood-brain barrier permeability and the brain extracellular space in acute cerebral inflammation.

The diffusion properties of the brain cortical extracellular space have never been examined in models of inflammation, even though inflammation can cause increased blood-brain barrier permeability. Uptake of intravascular 125I-labelled albumin and the diffusion of the tetramethylammonium ion within the brain extracellular space was measured in an experimental brain abscess to determine the effect of acute inflammation upon blood-brain barrier permeability and diffusion properties of the cortical extracellular space. The blood-brain transfer constant for albumin was increased in the abscess region, indicating that an increase in blood-brain barrier permeability occurred in animals inoculated with a weakly pathogenic strain of Staphylococcus aureus. The volume fraction of the extracellular space, as measured by the diffusion of tetramethylammonium ion, ranged from 0.19 to 0.23 in bacteria inoculated subjects and from 0.21 to 0.22 in controls. The tortuosity of the extracellular space ranged from 1.40 to 1.42 in bacteria inoculated subjects and was 1.39 in controls. These results showed that the volume fraction and tortuosity of the cortical extracellular space were not affected by inflammation even though vascular permeability was increased. This result was supported by the finding that brain water content, measured in the same animals, was increased to a non-significant extent in the bacteria inoculated subjects. These findings lead to the conclusion that acute inflammation induced by a weak pathogen can cause increased blood-brain barrier permeability without a significant change in the diffusion properties of the brain cortical space.

Animals↗