Recurring episodes of spreading depression are spontaneously elicited by an intracerebral hemorrhage in the swine.

Intracranial bleeding damages the surrounding tissue in a complex fashion that involves contamination by blood-borne products and loss of ionic homeostasis. We used electrophysiological techniques to examine the functional changes in the developing intracerebral bleed and in surrounding regions using an in vivo swine model. Intracerebral hemorrhage (ICH) was induced by collagenase injection into the primary somatosensory cortex (SI). Somatic evoked potential (SEP) elicited by electrical stimulation of the contralateral snout as well as changes in DC-coupled potential were monitored in the SI from the time of collagenase injection in order to measure the effects of ICH. The SEP decreased in amplitude within minutes of the intracerebral injection. Its short-latency component was abolished within the first hour after collagenase injection without any sign of recovery for the duration of the experiment. As the SEP started decreasing in amplitude, we observed spontaneous, recurring episodes of cortical spreading depression (SD) as early as 20 min post-injection. The timing of SDs in SI is consistent with our interpretation that SDs were initially generated at multiple sites adjacent to the lesion core and propagated into the surrounding area. With time, SD became less frequent near the injection site, shifting to more distant electrodes in the surrounding area. Our results indicate that ICH leads to the reduction in SEP amplitude and induces spontaneous episodes of SD. Loss of ionic homeostasis is most likely the physiological basis for the SEP change and for the induction of SD. Recurring SD spontaneously generated in experimental ICH needs further study in humans with ICH.

Roles of calcium- and voltage-sensitive potassium currents in the generation of neuromagnetic signals and field potentials in a CA3 longitudinal slice of the guinea-pig.

OBJECTIVES: Roles of calcium- and voltage-sensitive potassium currents in generation of neuromagnetic signals and field potentials were evaluated using the longitudinal CA3 slice preparation of the guinea-pig. METHODS: Their roles were evaluated by using selective channel blockers (tetraethyl-ammonium (TEA) and 4-aminopyridine (4AP)) and measuring their effects on the two types of signals and intracellular potentials. Fast gamma-aminobutyric acid type A inhibition was blocked with picrotoxin. RESULTS: Stimulation of the apical dendrites with an array of extracellular bipolar electrodes produced triphasic evoked magnetic fields with a spike and a slow wave typical of the slices. The evoked potentials in the apical and basal areas of the pyramidal cells closely resembled the magnetic field waveforms. Blockade of the potassium currents with TEA and 4AP had only subtle effects on the initial spike, but dramatically altered the slow wave. They also induced long-lasting spontaneous burst discharges synchronized across the slice. The results could be interpreted in terms of their known pre- and postsynaptic effects. Their post-synaptic effects were confirmed with intracellular recordings. CONCLUSION: Our results are consistent with a hypothesis that the calcium- and voltage-sensitive potassium currents, especially the A and C currents, play important roles in shaping the slow wave of the neuromagnetic and field potential signals produced by the mammalian hippocampus.

Roles of a potassium afterhyperpolarization current in generating neuromagnetic fields and field potentials in longitudinal CA3 slices of the guinea-pig.

OBJECTIVES: Roles of a calcium-dependent potassium conductance of slow afterhyperpolarization (AHP) type (gK(AHP)) in generating magnetoencephalographic (MEG) signals were studied in hippocampal longitudinal CA3 slices of the guinea pig. METHODS: The roles of gK(AHP) were experimentally inferred from effects of its blocker, carbamylcholine-chloride (carbachol, CCh), on MEG signals. The MEG signals were compared with extracellular field potentials and intracellular potentials of the pyramidal cells in the slice. RESULTS: CCh profoundly affected MEG waveforms. CCh reduced the initial spike of the evoked MEG signals independently of stimulation of the cell layer and apical dendrites. The slow wave of the evoked MEG signals was reduced by the somatic stimulation, but was enhanced by the apical stimulation. Elevated extracellular calcium and bath-applied tetraethylammonium (TEA) enhanced the CCh effects. CCh also increased spontaneous MEG signals. These effects on MEG and field potentials could be interpreted on the basis of synaptic and intracellular effects of CCh. CONCLUSIONS: Our results indicate that abnormality in this subtle calcium-dependent potassium channel may profoundly influence MEG and EEG signals.

Analysis of MEG signals of spreading cortical depression with propagation constrained to a rectangular cortical strip. I. Lissencephalic rabbit model.

Magnetic fields arising from the rabbit cortex during spreading cortical depression (SCD) were measured in order to study the currents in the neocortex during SCD. SCD was constrained to propagate in a rectangular cortical strip perpendicular to the midline. This simplified in vivo cortical preparation enabled us to correlate magnetoencephalographic (MEG) signals to their underlying currents within the cortical strip. The propagation of SCD was monitored with an array of electrodes placed along the strip. The propagation speed for SCD in the lissencephalic rabbit brain was 3. 5+/-0.3 mm/min (mean+/-S.E.M., n=14). Slow, quasi-dc, MEG signals were observed as the SCD entered into the longitudinal fissure. The currents giving rise to the MEG signals were perpendicular to the cortical surface and directed from the surface to deeper layers of the cortex. A distributed dipolar source model was used to relate the data to the underlying cortical current. The moment of the single equivalent current dipole source was 38+/-9 nA-m (n=17). This study clarified the nature of the cortical currents during SCD in a lissencephalic in vivo preparation.

Analysis of MEG signals of spreading cortical depression with propagation constrained to a rectangular cortical strip. II. Gyrencephalic swine model.

Currents produced during spreading cortical depression (SCD) in a gyrencephalic species (swine) were studied with magnetoencephalography (MEG) and electrocorticography (ECoG). SCD, initiated using electrical stimulation of the cortex, was constrained to propagate within a rectangular cortical strip in order to simplify the interpretation of the underlying currents. The ECoG signals monitored along the strip revealed that SCD propagated from an initiation site on the gyrus at a rate of 7.9+/-3.2 mm/min (n=23), entered the deep coronal sulcus and in most cases emerged from the other side of the sulcus, continuing to propagate across the next gyrus at a rate of 5.9+/-2.7 mm/min (n=22). The apparent propagation velocity within the sulcus was reduced to 1.7+/-0.8 mm/min (n=21). Strong MEG signals were observed as SCD entered the sulcus. The direction of magnetic field was opposite for SCD's on opposite banks of the sulcus. The currents were directed from a superficial layer to deeper layers of the cortex. The characteristics of SCD and associated MEG patterns from a gyrencephalic species may be similar to those in human patients during migraine aura.

Experimental analysis of distortion of magnetoencephalography signals by the skull.

OBJECTIVES: Magnetoencephalography (MEG) signals are, on theoretical grounds, thought to be relatively undistorted by the skull in contrast to electroencephalographic (EEG) signals. This assumption was experimentally tested in an animal preparation with a brain similar to the human brain in many respects. METHODS: Possible skull effects on MEG were evaluated directly using an in vivo porcine preparation, by measuring the somatic evoked magnetic field (SEF) above the skull with and without the skull under, otherwise, the same condition. RESULTS: The SEF was virtually undistorted by the skull with no obvious visible change in its waveform and amplitude under these two conditions. However, there was some small, but significant attenuation when the skull was removed, the distortion being greater for deeper sources. CONCLUSION: Our results are consistent with a theoretical expectation that the skull should be virtually 'transparent' to the magnetic fields for shallow sources, but less so for fields generated by deeper sources.

Roles of attention, memory, and motor preparation in modulating human brain activity in a spatial working memory task.

Neuronal activity of the human brain was studied with magnetoencephalography (MEG) in a spatial working memory task similar to those commonly used with nonhuman primates. The subject was required to remember target positions for 3 s and make a same-different judgement with a finger lift comparing the position of the probed target with the probe or to execute a memory-guided saccade to the probed target. In this type of task single-unit studies have shown attention- and memory-related activities independent of movement type during the retention interval in a large number of cortical areas of the primates, including the parietal and prefrontal areas. Consistent with these results, there were strong stimulus-driven transient and sustained responses and modulations of oscillatory activity during the retention period. Although we did not determine the source locations, coarse estimates of the currents responsible for the MEG signals showed activity over a wide area of the cortex, most prominently over the Rolandic, parietal and occipital areas, but also over the frontal area. Some of the activities in these cortical areas reflect processes that may be identified with attention and memory, while others were related to preparation of the overt movements.

Physiological bases of the synchronized population spikes and slow wave of the magnetic field generated by a guinea-pig longitudinal CA3 slice preparation.

OBJECTIVE: The physiological bases of evoked magnetic fields were examined in a guinea-pig hippocampal slice preparation, motivated by new concepts in central nervous system (CNS) electrophysiology brought about by discoveries of active conductances in the dendrites and soma of neurons. METHODS: Their origins were elucidated by comparing them with intracellular and extracellular field potentials. RESULTS: With excitatory synaptic transmissions blocked, the magnetic signal elicited by an electrical stimulus applied to the pyramidal cell layer consisted of a spike and a depolarizing afterpotential-like waveform. With the excitatory synaptic transmissions intact, but with inhibitory synaptic transmissions blocked, the magnetic signal was bi- or triphasic depending on whether the cell layer or the apical dendrite area of the pyramidal cells was, respectively, depolarized. In both cases the signal consisted of a train of synchronized population spikes superimposed on a brief wave followed by a longer, slow wave. The spike train was correlated with synaptically mediated intracellular spikes. The underlying currents for the slow wave were directed from the apical to the basal side for both types of stimulation. It was most likely generated by depolarization of the apical dendrites, caused by recurrent excitatory synaptic activation. CONCLUSIONS: This analysis illustrates how synaptic connections and intrinsic conductances in a disinhibited mammalian CNS structure can generate spikes and waves of the magnetic field and electrical potential.

Genesis of MEG signals in a mammalian CNS structure.

Neuromagnetic signals of guinea pig hippocampal slices were characterized and compared with the extracellular field potential to elucidate the genesis of magnetoencephalographic signals in a mammalian CNS structure. The spatial distribution of magnetic evoked field (MEF) directed normal to bath surface was similar for transverse CA1, longitudinal CA1 and longitudinal CA3 slices in the presence of 0.1 mM picrotoxin (PTX) which blocks inhibitory synaptic transmission. Their MEFs were produced by currents along the longitudinal axis of the pyramidal cells. Comparisons of the MEF with the laminar potential profile revealed that the MEF was generated by intracellular longitudinal currents. The dipolar component of the intracellular currents was the dominant factor generating the MEF even at a distance of 2 mm from the slice. The MEF from a slice in Ringer's solution without PTX became similar in temporal waveform with time to the MEF in the presence of PTX, indicating the predominance of excitatory connections in generating the MEF and the existence of highly synchronous populations activities across the slice even in PTX-free Ringer's solution. The presence of such highly synchronous population activities underlying the MEFs was verified directly with field potentials recorded across the slice. A systematic variation of the stimulation site revealed a characteristic waveform for each site. The variation of the with stimulation site suggested the contribution of many factors, both synaptic and voltage-sensitive conductances, to the overall waveform of the MEF.

Single-epoch neuromagnetic signals during epileptiform activities in guinea pig longitudinal CA3 slices.

Neuromagnetic fields with high signal-to-noise ratios can be measured above longitudinal CA3 slices of the guinea pig during single epochs of interictal- and ictal-like synchronized population activities. Technical refinements enabled us to reduce the number of epochs for clear responses from over 5000 needed in an earlier study to single epochs. Simultaneous recording of field potentials revealed that neuromagnetic fields reflect intracellular currents in the pyramidal cells. Intracellular currents could be estimated from the external magnetic fields during paroxysmal depolarization shifts, multiple bursts, and slowly varying potential shifts lasting several seconds.

Neuronal activities from a deep subcortical structure can be detected magnetically outside the brain in the porcine preparation.

We assessed whether subcortical structures can generate magnetic fields detectable outside the brain by first measuring the somatic evoked magnetic fields (SEFs) from a decorticated porcine preparation and then from an intact preparation. Strong SEFs were detected a few millimeters above the corpus callosum after electrical stimulations of the snout. The waveforms consisted of a large spike ( < or = 6 pT) with a peak latency of 11-18 ms depending on the age of the animal, followed by a slow wave. The waveform and latency of the SEF spike were virtually identical to those of the field potential within the brain. The SEF topography indicated that the underlying generator of the spike was in a region contralateral to the stimulation and inferior to the thalamus. The subcortical SEF was strong enough to be detectable even above the intact brain, after the cortically generated SEF was removed by ablation of the primary cortical area. The results indicate that a structure deep in the brain can produce remarkably strong magnetic fields detectable outside the brain.

Origin of the apparent tissue conductivity in the molecular and granular layers of the in vitro turtle cerebellum and the interpretation of current source-density analysis.

1. We determined the origin of the apparent tissue conductivity (sigma 2) of the turtle cerebellum in vitro. 2. Application of a current with a known current density (J) along the longitudinal axis of a conductivity cell produced an electric field in the cerebellum suspended in the cell. The measured electric field (E) perpendicular to the cerebellar surface indicated a significant inhomogeneity in sigma a (= J/E) with a major discontinuity between the molecular layer (0.25 +/- 0.05 S/m, mean +/- SD) and granular layers (0.15 +/- 0.03 S/m) (n = 39). 3. This inhomogeneity was more pronounced after anoxic depolarization. The value of sigma a decreased to 0.11 +/- 0.03 and 0.040 +/- 0.008 S/m in the molecular and granular layers, respectively. The ratio of sigma a S in the two layers increased from 1.67 in the normoxic condition to 2.75 after anoxic depolarization. 4. This difference in sigma a across the two layers was present within the range of frequencies (DC to 10 kHz) studied where the phase of sigma a was small (less than +/- 2 degrees) and therefore sigma a was ohmic. 5. The inhomogeneity in sigma a was in part due to an inhomogeneity in the extracellular conductivity (sigma e) as determined from the extracellular diffusion of ionophoresed tetramethylammonium. Like sigma a, the value of sigma e was also higher in the molecular layer (0.165 S/m) than in the granular layer (0.097 S/m). The inhomogeneity in sigma e was due to a smaller tortuosity and a larger extracellular volume fraction in the molecular layer compared with the granular layer. 6. sigma a was, however, consistently higher, by approximately 50%, than sigma e. A core conductor model of the cerebellum indicated that these discrepancies between sigma a and sigma e were attributable to additional conductivity produced by a passage of the longitudinal applied current through the intracellular space of Purkinje cells and ependymal glial cells, with the glial compartment playing the dominant role. Cells with a long process and a short space constant such as the ependymal glia evidently enhance the effective "extracellular" conductivity by serving as intracellular conduits for the applied current. The result implies that the effective sigma e may be larger than sigma e for neuronally generated currents in the turtle cerebellum because the space constant for Purkinje cells is several times greater than that for the ependymal glia and consequently Purkinje cell-generated currents travel over a long distance relative to the space constant of glial cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Ferromagnetic high-permeability alloy alone can provide sufficient low-frequency and eddy-current shieldings for biomagnetic measurements.

A low cost, two-shell ferromagnetic shielded room large enough for a person to enter and prepare experiments was constructed for biomagnetic measurements. No aluminium or copper shell was used for eddy-current shielding. A high-permeability nickel-iron-molybdenum alloy (1.57 mm thick) was used for both ferromagnetic and eddy-current shieldings. The shielding factor was 60 dB at dc and 40 dB between 0.1 Hz and about 3 Hz. The eddy-current shielding due to the alloy alone provided a shielding factor of 55 dB at 30 Hz and 66 dB at 60 Hz. The shielding factor was sufficiently high in all the frequency range for biomagnetic measurements with a first-order superconducting gradiometer.

Detection of magnetic evoked fields associated with synchronous population activities in the transverse CA1 slice of the guinea pig.

1. Magnetic evoked fields (MEFs) associated with synchronous population activities in the transverse CA1 of guinea pig (approximately 1.2 mm3) were characterized with a high-resolution superconducting magnetic field detector. 2. Electrical stimulations of the stratum radiatum with a pair of bipolar electrodes produced synchronous population spikes in the field potential and a strong MEF (as much as 3.5 picoteslas 3 mm above tissue) in the presence of 0.05 mM picrotoxin. The MEF's spatial pattern and direction indicated currents in pyramidal cells as its source. Bath application of kynurenic acid extinguished most of the signals, indicating a strong contribution of post-synaptic currents to the field. The kynurenic acid-insensitive component was abolished by tetrodotoxin, indicating the remaining component was neuronal in origin as well. 3. It may be possible to refine our technique to study the genesis of MEG signals--i.e., the relationship between evoked fields and the underlying neuronal currents--in a mammalian CNS structure, since the electrophysiology of the hippocampus is well understood.

Anisotropic and heterogeneous diffusion in the turtle cerebellum: implications for volume transmission.

1. Measurements of extracellular diffusion properties were made in three orthogonal axes of the molecular and granular layers of the isolated turtle cerebellum with the use of iontophoresis of tetramethylammonium (TMA+) combined with ion-selective microelectrodes. 2. Diffusion in the extracellular space of the molecular layer was anisotropic, that is, there was a different value for the tortuosity factor, lambda i, associated with each axis of that layer. The x- and y-axes lay in the plane parallel to the pial surface of this lissencephalic cerebellum with the x-axis in the direction of the parallel fibers. The z-axis was perpendicular this plane. The tortuosity values were lambda x = 1.44 +/- 0.01, lambda y = 1.95 +/- 0.02, and lambda z = 1.58 +/- 0.01 (mean +/- SE). By contrast, the granular layer was isotropic with a single tortuosity value, lambda Gr = 1.77 +/- 0.01. 3. These data confirm the applicability of appropriately extended Fickian equations to describe diffusion in anisotropic porous media, including brain tissue. 4. Heterogeneity between the molecular and granular layer was revealed by a striking difference in extracellular volume fraction, alpha, for each layer. In the molecular layer alpha = 0.31 +/- 0.01, whereas in the granular layer alpha = 0.22 +/- 0.01. 5. Volume fraction and tortuosity affected the time course and amplitude of extracellular TMA+ concentration after iontophoresis. This was modeled by the use of the average parameters determined experimentally, and the nonspherical pattern of diffusion in the molecular layer was compared with the spherical distribution in the granular layer and agarose gel by computing isoconcentration ellipsoids. 6. One functional consequence of these results was demonstrated by measuring local changes in [K+]o and [Ca2+]o after microiontophoresis of a cerebellar transmitter, glutamate. The ratios of ion shifts in the x- and y-axes in the granular layer were close to unity, with a ratio of 1.04 +/- 0.08 for the rise in [K+]o and 1.03 +/- 0.17 for the decrease in [Ca2+]o. In contrast, ion shifts in the molecular layer had an x:y ratio of 1.44 +/- 0.14 for the rise in [K+]o and 2.10 +/- 0.42 for the decrease in [Ca2+]o. 7. These data demonstrate that the structure of cellular aggregates can channel the migration of substances in the extracellular microenvironment, and this could be a mechanism for volume transmission of chemical signals. For example, the preferred diffusion direction of glutamate along the parallel fibers would help constrain an incoming excitatory stimulus to stay "on-beam."

Multimodal characterization of population responses evoked by applied electric field in vitro: extracellular potential, magnetic evoked field, transmembrane potential, and current-source density analysis.

An external electric field applied parallel to longitudinal axis of neurons selectively depolarizes either end and thereby activates voltage-sensitive conductance changes in a large population of neurons. Here, we characterized such population responses in the in vitro turtle cerebellum. The responses were recorded and analyzed using a multimodal approach: the magnetic evoked field was measured using a Superconducting Quantum Interference Device (SQUID) magnetometer, and concurrently the electric field potentials were recorded. Laminar profile and current-source density analysis were used to uncover the pattern of activation due to the applied electric field. Intracellular recording provided further information for identifying the elements producing the observed responses. Finally, pharmacological manipulations confirmed the nature of the conductance changes. Our results show that it is possible to activate a defined cell population of the cerebellum by an applied field and obtain a magnetic response of the order of 0.5-2 pT. A field applied from the dorsal to the ventral side of cerebellum produced tetrodotoxin-sensitive population spikes. This component was followed by a kynurenic acid (KYNA)-sensitive postsynaptic response, most likely comprised of Ca(2+)-mediated action potentials occurring at the proximal pole of the Purkinje cell dendrites and evoked by climbing fiber inputs. The applied electric field directed from the ventral to the dorsal side of cerebellum gave rise to a complex of responses that was made up of a KYNA-sensitive component (presumably synaptically activated) and an Mn(2+)-sensitive but KYNA-insensitive component (probably due to a directly activated calcium conductance change). This study provides insights into the effects of electric and magnetic fields applied to the nervous tissue of experimental animal and human studies.

Distortion of magnetic evoked fields and surface potentials by conductivity differences at boundaries in brain tissue.

We investigated the conditions under which inhomogeneity in electrical conductivity may significantly modify the magnetic evoked field (MEF) due to primary currents (i.e., neuronal currents) in the brain. In the case of an isolated turtle cerebellum immersed in a large bath of physiological saline, our theoretical analysis showed the cerebellar surface to significantly enhance the MEF due to a primary current, by a factor of as much as two, for experimentally determined values of the conductivities of the cerebellar tissue and saline. A further parametric investigation of the conductivity effect revealed that conductivity boundaries may significantly modify the MEF due to neuronal currents located within 1 mm of a conductivity boundary, as would be the case for active neurons near an edema, an anoxic fringe such as might occur during stroke, or a ventricle in the human head. For a stationary neural source, conductivity boundaries may modify the magnitude of its MEF without affecting its temporal waveform. However, this boundary effect was found to be small for a model geometry locally approximating cortical sources in a sulcus or a fissure, where the boundary effects from adjacent sulcal walls tend to cancel each other.

Magnetic evoked field associated with transcortical currents in turtle cerebellum.

The neural basis of magnetic evoked fields of the brain was studied with an isolated turtle cerebellum as a model preparation. The turtle cerebellum is a nearly flat tissue with neural processes arranged along three orthogonal axes of symmetry. According to theoretical results, this geometry should enable us to selectively measure the magnetic field due to a subpopulation of nerve cells whose longitudinal axes are perpendicular to the cerebellar surface, by simply placing the cerebellum vertically in the bath so that these cells are horizontal and by measuring the field along the rostrocaudal axis perpendicular to the longitudinal axis of these nerve cells. To elicit neural activity in these cells the dorsal midline was electrically stimulated with a bipolar electrode. Consistent with our expectations, the one-dimensional profile of the field normal to bath surface (Bz) was antisymmetrical along the rostrocaudal axis, implying that the underlying currents were transcortical. Also, the Bz field at a field extremum varied as a cosine of the orientation of the cerebellum when it was rotated about its rostrocaudal axis with the amplitude being zero when the cerebellum was horizontal. The Bz field was dipolar as judged by statistically excellent fits of the dipolar field to the one-dimensional field profile and to the distance function relating the field magnitude at an extremum to measuring distance. This was the case even for the initial component thought to be due to antidromic action currents invading the soma and dendrites of Purkinje cells. We also showed that the dipolar term of the source could be localized within 1 mm of the actual source location in most cases.

Magnetic field associated with spreading depression: a model for the detection of migraine.

Slow variations of the magnetic field were recorded in real time during spreading depression (SD) in the isolated turtle cerebellum. The magnetic signal lasted for 2-10 min with the largest amplitude in the first minute. The field strength was of sufficient magnitude to be measured unaveraged at 2-4 cm from the tissue. The directions and time course of the magnetic signal indicated that cerebellar SD is accompanied by current normal to the cerebellar surface. The observations reported here are of clinical interest due to the potential involvement of SD in various neurological disorders, notably head trauma and migraine.