Multi-sensor magnetoencephalography with atomic magnetometers.

The authors have detected magnetic fields from the human brain with two independent, simultaneously operating rubidium spin-exchange-relaxation-free magnetometers. Evoked responses from auditory stimulation were recorded from multiple subjects with two multi-channel magnetometers located on opposite sides of the head. Signal processing techniques enabled by multi-channel measurements were used to improve signal quality. This is the first demonstration of multi-sensor atomic magnetometer magnetoencephalography and provides a framework for developing a non-cryogenic, whole-head magnetoencephalography array for source localization.

Low power high-performance radio frequency oscillator for driving ion traps.

We report a simple, efficient, high voltage radio frequency (RF) generator powered by a single voltage source (1.5-7 V) to resonantly drive ion traps or other capacitive loads. Our circuit is able to deliver RF voltages > 500 V(p-p) at frequencies ranging from 0.1 to 10 MHz. This RF oscillator uses low-cost, commercially available components, and can be easily assembled onto a circuit board of a few cm(2). Because of its simplicity and good efficiency, this circuit is useful in applications requiring small size and low power consumption such as portable ion trap systems where the duration of operation under battery power is of concern.

Compact phase delay technique for increasing the amplitude of coherent population trapping resonances in open Lambda systems.

We propose and demonstrate a novel technique for increasing the amplitude of coherent population trapping (CPT) resonances in open Lambda systems. The technique requires no complex modifications to the conventional CPT setup and is compatible with standard microfabrication processes. The improvement in the CPT resonance amplitude as a function of intensity of the excitation light agrees well with the theory based on ideal open and closed Lambda systems.

Atomic vapor cells for chip-scale atomic clocks with improved long-term frequency stability.

A novel technique for microfabricating alkali atom vapor cells is described in which alkali atoms are evaporated into a micromachined cell cavity through a glass nozzle. A cell of interior volume 1 mm3, containing 87Rb and a buffer gas, was made in this way and integrated into an atomic clock based on coherent population trapping. A fractional frequency instability of 6 x 10(-12) at 1000 s of integration was measured. The long-term drift of the F=1, mF=0-->F=2, mF=0 hyperfine frequency of atoms in these cells is below 5 x 10(-11)/day.

A chip-scale atomic clock based on 87Rb with improved frequency stability.

We demonstrate a microfabricated atomic clock physics package based on coherent population trapping (CPT) on the D1 line of 87Rb atoms. The package occupies a volume of 12 mm3 and requires 195 mW of power to operate at an ambient temperature of 200 degrees C. Compared to a previous microfabricated clock exciting the D2 transition in Cs [1], this 87Rb clock shows significantly improved short- and long-term stability. The instability at short times is 4 x?10-11 / tau?/2 and the improvement over the Cs device is due mainly to an increase in resonance amplitude. At longer times (tau?> 50 s), the improvement results from the reduction of a slow drift to ?5 x 10-9 / day. The drift is most likely caused by a chemical reaction of nitrogen and barium inside the cell. When probing the atoms on the D1 line, spin-exchange collisions between Rb atoms and optical pumping appear to have increased importance compared to the D2 line.

Initiation and propagation of regenerative Ca(2+)-dependent potentials in dendrites of layer 5 pyramidal neurons.

The initiation and propagation of dendritic Ca(2+)-dependent regenerative potentials (CDRPs) were investigated by imaging the Ca(2+)-sensitive dye Fluo-4 during whole cell recording from the soma of layer 5 pyramidal neurons visualized in a slice preparation of rat neocortex by the use of infrared-differential interference contrast microscopy. CDRPs were evoked by focal iontophoresis of glutamate at visually identified sites 178-648 microm from the soma on the apical dendrite and at sites on the basal dendrites. Increases in [Ca(2+)](i) were maximal near the site of iontophoresis and were graded with iontophoretic current that was subthreshold for evoking CDRPs. CDRP initiation was associated with a [Ca(2+)](i) rise that differed from a just-subthreshold response in both magnitude and spatial extent but whose amplitude declined both proximal and distal to the iontophoretic site. These [Ca(2+)](i) rises, whether associated with subthreshold or regenerative voltage responses, were minimally affected by blockade of N-methyl-D-aspartate receptors but were abolished by Cd(2+), suggesting that Ca(2+) influx through voltage-gated channels caused the rise of [Ca(2+)](i). On the assumption that the rise of [Ca(2+)](i) during a CDRP marks the spatial extent of regenerative Ca(2+) influx, we conclude that CDRPs can be evoked at any point on the main apical or basal trunk where membrane potential reaches CDRP threshold rather than at discrete "hot spots," the CDRP is initiated at a spatially restricted site, and it propagates decrementally both distal and proximal to its initiation site. These results raise the possibility that synaptic integration may occur first in the dendrites to evoke a CDRP. Because these responses propagate decrementally to the soma, they are able to sum with input from other regions of the cell so that the cell as a whole remains integrative.

Dendritic calcium spikes in layer 5 pyramidal neurons amplify and limit transmission of ligand-gated dendritic current to soma.

Long-lasting, dendritic, Ca(2+)-dependent action potentials (plateaus) were investigated in layer 5 pyramidal neurons from rat neocortical slices visualized by infrared-differential interference contrast microscopy to understand the role of dendritic Ca(2+) spikes in the integration of synaptic input. Focal glutamate iontophoresis on visualized dendrites caused soma firing rate to increase linearly with iontophoretic current until dendritic Ca(2+) responses caused a jump in firing rate. Increases in iontophoretic current caused no further increase in somatic firing rate. This limitation of firing rate resulted from the inability of increased glutamate to change evoked plateau amplitude. Similar nonlinear patterns of soma firing were evoked by focal iontophoresis on the distal apical, oblique, and basal dendrites, whereas iontophoresis on the soma and proximal apical dendrite only evoked a linear increase in firing rate as a function of iontophoretic current without plateaus. Plateau amplitude recorded in the soma decreased as the site of iontophoresis was moved farther from the soma, consistent with decremental propagation of the plateau to the soma. Currents arriving at the soma summed if plateaus were evoked on separate dendrites or if subthreshold responses were evoked from sites on the same dendrite. If plateaus were evoked at two sites on the same dendrite, only the proximal plateau was seen at the soma. Just-subthreshold depolarizations at two sites on the same dendrite could sum to evoke a plateau at the proximal site. We conclude that the plateaus prevent current from ligand-gated channels distal to the plateau-generating region from reaching the soma and directly influencing firing rate. The implications of plateau properties for synaptic integration are discussed.

Cytogenetic and molecular characterization of a patient with simultaneous B-cell chronic lymphocytic leukemia and peripheral T-cell lymphoma.

A patient is described who developed a peripheral T-cell lymphoma (PTCL) after a 6-year history of B-cell chronic lymphocytic leukemia (B-CLL). The progression of the T-cell disease spreading to pleura and skin terminated the course of the disease. A cytogenetic analysis performed six years after the first onset of the B-CLL showed the presence of two clones, one with trisomy 12 and another with inv(14)(q11q32.1) and trisomy 8. Combined immunophenotyping and fluorescence in situ hybridization demonstrated that only CD19+ cells contained a trisomy 12, whereas CD3+ cells contained a trisomy 8. Analyses of IgH and TCR rearrangements in single micromanipulated B- and T-cells lacked evidence for a clonal relation between B-CLL and PTCL cells. Based on our findings, we discuss the different hypotheses which might explain the development of simultaneous PTCL and B-CLL.

Waveguide atom beam splitter for laser-cooled neutral atoms.

A laser-cooled neutral-atom beam from a low-velocity intense source is split into two beams while it is guided by a magnetic-field potential. We generate our multimode beam-splitter potential with two current-carrying wires upon a glass substrate combined with an external transverse bias field. The atoms are guided around curves and a beam-splitter region within a 10-cm guide length. We achieve a maximum integrated flux of 1.5x10(5)atoms/s with a current density of 5x10(4)amp/cm (2) in the 100-microm -diameter wires. The initial beam can be split into two beams with a 50/50 splitting ratio.

Mechanisms underlying burst and regular spiking evoked by dendritic depolarization in layer 5 cortical pyramidal neurons.

Apical dendrites of layer 5 pyramidal cells in a slice preparation of rat sensorimotor cortex were depolarized focally by long-lasting glutamate iontophoresis while recording intracellularly from their soma. In most cells the firing pattern evoked by the smallest dendritic depolarization that evoked spikes consisted of repetitive bursts of action potentials. During larger dendritic depolarizations initial burst firing was followed by regular spiking. As dendritic depolarization was increased further the duration (but not the firing rate) of the regular spiking increased, and the duration of burst firing decreased. Depolarization of the soma in most of the same cells evoked only regular spiking. When the dendrite was depolarized to a critical level below spike threshold, intrasomatic current pulses or excitatory postsynaptic potentials also triggered bursts instead of single spikes. The bursts were driven by a delayed depolarization (DD) that was triggered in an all-or-none manner along with the first Na+ spike of the burst. Somatic voltage-clamp experiments indicated that the action current underlying the DD was generated in the dendrite and was Ca2+ dependent. Thus the burst firing was caused by a Na+ spike-linked dendritic Ca2+ spike, a mechanism that was available only when the dendrite was adequately depolarized. Larger dendritic depolarization that evoked late, constant-frequency regular spiking also evoked a long-lasting, Ca2+-dependent action potential (a "plateau"). The duration of the plateau but not its amplitude was increased by stronger dendritic depolarization. Burst-generating dendritic Ca2+ spikes could not be elicited during this plateau. Thus plateau initiation was responsible for the termination of burst firing and the generation of the constant-frequency regular spiking. We conclude that somatic and dendritic depolarization can elicit quite different firing patterns in the same pyramidal neuron. The burst and regular spiking observed during dendritic depolarization are caused by two types of Ca2+-dependent dendritic action potentials. We discuss some functional implications of these observations.

Synaptically evoked dendritic action potentials in rat neocortical pyramidal neurons.

In a previous study iontophoresis of glutamate on the apical dendrite of layer 5 pyramidal neurons from rat neocortex was used to identify sites at which dendritic depolarization evoked small, prolonged Ca2+ spikes and/or low-threshold Na+ spikes recorded by an intracellular microelectrode in the soma. These spikes were identified as originating in the dendrite. Here we evoke similar dendritic responses by electrical stimulation of presynaptic elements near the tip of the iontophoretic electrode with the use of a second extracellular electrode. In 9 of 12 recorded cells, electrically evoked excitatory postsynaptic potentials (EPSPs) above a minimum size triggered all-or-none postsynaptic responses similar to those evoked by dendritic glutamate iontophoresis at the same site. Both the synaptically evoked and the iontophoretically evoked depolarizations were abolished reversibly by blockade of glutamate receptors. In all recorded cells, the combination of iontophoresis and an EPSP, each of which was subthreshold for the dendritic spike when given alone, evoked a dendritic spike similar to that evoked by a sufficiently large iontophoresis. In one cell tested, dendritic spikes could be evoked by the summation of two independent subthreshold EPSPs evoked by stimulation at two different locations. We conclude that the dendritic spikes are not unique to the use of glutamate iontophoresis because similar spikes can be evoked by EPSPs. We discuss the implications of these results for synaptic integration and for the interpretation of recorded synaptic potentials.

Reduction of cortical pyramidal neuron excitability by the action of phenytoin on persistent Na+ current.

We examined the effect of the anticonvulsant phenytoin (PT) (20-200 microM) on the persistent Na+ current (INaP), INaP-dependent membrane potential responses and repetitive firing in layer 5 pyramidal neurons in a slice preparation of rat sensorimotor cortex. INaP measured directly with voltage-clamp was reduced in a concentration-dependent manner with an apparent EC50 value of 78 microM. Clear effects on current-evoked membrane potential responses were apparent at 50 microM PT: Subthreshold, depolarizing membrane potential rectification was reduced, rheobase current was increased and the relation between firing rate and injected current was shifted to the right, but action potential amplitude and duration were unaffected. We ascribed these effects of PT largely to the reduction of INaP. A slow decline of firing rate during the injected current pulse also became apparent at moderate PT concentrations. When PT concentration was raised to 150 to 200 microM, this slow adaption was enhanced markedly, and firing ceased during a sufficiently large current pulse. This enhanced slow adaptation and the cessation of firing were associated with a marked decline of spike amplitude and a rise in spike firing level during successive interspike intervals. We ascribe these effects largely to the action of PT on the transient Na+ current. We conclude that the reduction in cortical neuronal excitability by PT depends partly on its reduction of INaP, the effects of INaP blockade are apparent at PT concentrations lower than those required to abolish tonic firing and the cells need not be excessively depolarized for PT to decrease excitability by its effect on INaP.

Evidence for persistent Na+ current in apical dendrites of rat neocortical neurons from imaging of Na+-sensitive dye.

Evidence for a persistent Na+ current (I(NaP)) in the apical dendrite of neocortical neurons was sought with the use of fluorescence imaging to measure changes in intradendritic Na+ concentration. Neurons in neocortical brain slices were filled iontophoretically through an intracellular recording microelectrode with the Na+-sensitive dye benzofuran isophthalate (SBFI), and fluorescence images were recorded with a cooled charge-coupled device camera system using 380-nm illumination. In the presence of Ca2+ and K+ channel blockers, a short depolarizing current pulse evoked a single action potential followed by a plateau depolarization (PD) lasting >1 s. This tetrodotoxin (TTX)-sensitive PD is known to be maintained by I(NaP). A single action potential caused no detectable SBFI fluorescence change, whereas the PD was associated with an SBFI fluorescence change in the soma and apical dendrite indicating increased intracellular Na+ concentration. Determination of the full spatial extent of the dendritic fluorescence change was prevented by our inability to detect the dim fluorescence signal in the distal regions of the apical dendrite. In each experiment the fluorescence change extended into the apical dendrite as far as dye could be visualized (50-300 microm). A slow, depolarizing voltage-clamp ramp that activated I(NaP) caused similar fluorescence changes that were eliminated by TTX, indicating that the SBFI fluorescence changes are caused by Na+ influx due to I(NaP) activation. We conclude that I(NaP) can be generated by the apical dendritic membrane to at least 300 microm from the soma.

Modification of current transmitted from apical dendrite to soma by blockade of voltage- and Ca2+-dependent conductances in rat neocortical pyramidal neurons.

The axial current transmitted to the soma during the long-lasting iontophoresis of glutamate at a distal site on the apical dendrite was measured by somatic voltage clamp of rat neocortical pyramidal neurons. Evidence for voltage- and Ca2+-gated channels in the apical dendrite was sought by examining the modification of this transmitted current resulting from the alteration of membrane potential and the application of channel-blocking agents. After N-methyl-D-aspartate receptor blockade, iontophoresis of glutamate on the soma evoked a current whose amplitude decreased linearly with depolarization to an extrapolated reversal potential near 0 mV. Under the same conditions, glutamate iontophoresis on the apical dendrite 241-537 microm from the soma resulted in a transmitted axial current that increased with depolarization over the same range of membrane potential (about -90 to -40 mV). Current transmitted from dendrite to soma was thus amplified during depolarization from resting potential (about -70 mV) and attenuated during hyperpolarization. After Ca2+ influx was blocked to eliminate Ca2+-dependent K+ currents, application of 10 mM tetraethylammonium chloride (TEA) altered the amplitude and voltage dependence of the transmitted current in a manner consistent with the reduction of dendritic voltage-gated K+ current. We conclude that dendritic, TEA-sensitive, voltage-gated K+ channels can be activated by tonic dendritic depolarization. The most prominent effects of blocking Ca2+ influx resembled those elicited by TEA application, suggesting that these effects were caused predominantly by blockade of a dendritic Ca2+-dependent K+ current. When cells were impaled with microelectrodes containing ethylene glycol-bis(beta-amino-ethyl ether)-N,N',N'-tetraacetic acid to prevent a rise in intracellular Ca2+ concentration, blockade of Ca2+ influx altered the tonic transmitted current in different manner consistent with the blockade of an inward dendritic current carried by high-threshold-activated Ca2+ channels. We conclude that the primary effect of Ca2+ influx during tonic dendritic depolarization is the activation of a dendritic Ca2+-dependent K+ current. The hyperpolarizing attenuation of transmitted current was unaffected by blocking all known voltage-gated inward currents except the hyperpolarization-activated cation current (Ih). Extracellular Cs+ (3 mM) reversibly abolished both the hyperpolarizing attenuation of transmitted current and Ih measured at the soma. We conclude that activation of Ih by hyperpolarization of the proximal apical dendrite would cause less axial current to arrive at the soma from a distal site than in a passive dendrite. Several functional implications of dendritic K+ and Ih channels are discussed.

Local and propagated dendritic action potentials evoked by glutamate iontophoresis on rat neocortical pyramidal neurons.

Iontophoresis of glutamate at sites on the apical dendrite 278-555 microm from the somata of rat neocortical pyramidal neurons evoked low-threshold, small, slow spikes and/or large, fast spikes in 71% of recorded cells. The amplitude of the small, slow spikes recorded at the soma averaged 9.1 mV, and their apparent threshold was <10 mV positive to resting potential. Both their amplitude and their apparent threshold decreased as the iontophoretic site was moved farther from the soma. These spikes were not abolished by somatic hyperpolarization. When the somata of cells displaying these small spikes were voltage clamped at membrane potentials that prevented somatic or axonic firing, corresponding current spikes could be evoked all-or-none by dendritic depolarization, indicating that the small, slow spikes arose in the dendrite. Similar responses were not observed during somatic depolarization evoked by current pulses or glutamate iontophoresis. These small, slow spikes were abolished by blocking voltage-gated Ca2+ channels but not by blocking Na+ channels or N-methyl-D-aspartate receptors. We conclude that these Ca2+ spikes occurred in a spatially restricted region of the dendrite and were not actively propagated to the soma. In the presence of 10 mM tetraethylammonium chloride, the amplitudes of the iontophoretically evoked Ca2+ spikes were large, similar to those of the Ca2+ spikes evoked by somatic current injection, but their apparent thresholds were 63% lower. We conclude that dendritic K+ channels normally prevent the active propagation of Ca2+ spikes along the dendrite. In 36% of recorded cells dendritic glutamate iontophoresis evoked a Na+ spike with an apparent threshold 63% lower than those evoked by somatic current injection or somatic glutamate iontophoresis. Blockade of these low-threshold Na+ spikes by pharmacological or electrophysiological means often revealed underlying small dendritic Ca2+ spikes. When cells displaying the low-threshold Na+ spikes were voltage clamped at membrane potentials that prevented firing of the soma or axon, corresponding tetrodotoxin-sensitive current spikes could be evoked all-or-none by dendritic depolarization. We conclude that these low-threshold Na+ spikes were initiated in the dendrite, probably by local Ca2+ spikes, and subsequently propagated actively to the soma. Most cells displaying dendritic Na+ spikes fired multiple bursts of action potentials during tonic dendritic depolarization, whereas somatic depolarization of the same cells evoked only regular firing. We discuss the implications of dendritic Ca2+ and Na+ spikes for synaptic integration and neural input-output relations.

Quantitative analysis of firing properties of pyramidal neurons from layer 5 of rat sensorimotor cortex.

Quantitative aspects of repetitive firing evoked by injected current steps and ramps were studied in layer 5 pyramidal neurons in brain slices of rat sensorimotor cortex to answer the following questions. Do the tonic firing properties of burst-firing and regular-spiking (nonbursting) neurons differ significantly? Does burst firing denote a discrete class of neurons or represent a continuum of firing properties? Is firing rate during the burst of action potentials related to stimulus amplitude? What aspect of the stimulus might the initial firing rate code? How stable are a neuron's firing properties over time? All recorded neurons fired tonically to a long-lasting current above a minimum value, and the tonic firing properties of most neurons were quite similar irrespective of their initial response to a current step. Only a group of high-resistance neurons had significantly different tonic firing properties. When slow current ramps (rising between 0.5 and approximately 20 nA/s) were applied, the relation between firing rate and current during the ramp was very similar to the relation between tonic firing rate and current obtained from long-lasting current steps. Low-resistance cells exhibited three distinct initial responses to a current step: fast adaptation, high-threshold bursts, and low-threshold bursts, observed in 54, 28, and 10% of recorded cells, respectively. High-resistance cells exhibited a distinctive slow adaptation of firing rate. Slowly adapting, fast-adapting (FA), and high-threshold burster (HTB) neurons exhibited no adaptation near the minimum current that evoked repetitive firing (I(o)). FA and HTB cells exhibited two-spike adaptation to a fina tonic firing rate during currents up to 1.6 times I(o). Only a higher current (2.1 times I(o)) evoked a burst in HTB cells, whereas a burst was evoked at I(o) in the low-threshold burster cells. In most cells analyzed, the initial firing rate, whatever its nature, increased monotonically with current step amplitude. The response to fast current ramps indicated that firing rate during adaptation or bursting may code rate of change of current. Repeated measurements during long-duration impalements indicated that both transient and tonic firing properties are stable over time. We discuss how the different tonic firing properties of large and small pyramidal neurons could be more important functionally than the different transient responses (burst/nonburst) of the large neurons. We conclude that the large neurons would perform a better linear transduction of time-varying synaptic current that reaches their somata. We compare the responses evoked by somatically injected current with those evoked by dendritic glutamate iontophoresis in previous studies.

Equivalence of amplified current flowing from dendrite to soma measured by alteration of repetitive firing and by voltage clamp in layer 5 pyramidal neurons.

1. Plots of steady firing rate versus injected current (f-I relations) were constructed from intrasomatic injected current pulses applied alone (control relations) and together with dendritic glutamate iontophoresis (test relations) at sites on the distal apical dendrite 185-555 microns from the soma in layer 5 pyramidal neurons from rat cortex studied in a brain slice. The test f-I relations exhibited a parallel shift along the current axis, and the slopes of the control and test relations differed by < 10% in most neurons. This behavior indicates that constant injected current and steady glutaminergic dendritic input evoke equivalent steady-state repetitive firing in a neuron with active dendrites. The parallel shift of the f-I curves allowed us to compute the amplitude of axial current arriving in the soma from the apical dendrite during repetitive firing. 2. We compared the transmitted current computed from the f-I curve shift with that measured by somatic voltage clamp during the same iontophoresis. When measured during voltage clamp at different somatic membrane potentials, the transmitted current increased with somatic depolarization (was amplified) in most cells, an observation inconsistent with passive dendrites. This larger-amplitude current closely predicted the transmitted current computed from the f-I curve shift, whereas the smaller transmitted current measured at resting potential did not. A set of control experiments indicated that these different predictions were well within the measurement error associated with computation of transmitted current based on f-I curve shifts. The action of blocking agents confirmed that the depolarizing amplification depended on tetrodotoxin (TTX)- and D-2-amino-5-phosphonopentoic acid (APV)-sensitive dendritic channels. 3. The agreement of two independent measurements (somatic voltage clamp and f-I curve shift) of the axial current transmitted from dendrite to soma indicates that the amplification of transmitted current observed in voltage clamp occurs physiologically. We discuss the usefulness of the effective current concept for determining synaptic weighting in network models of neurons with active dendrites.

Amplification of synaptic current by persistent sodium conductance in apical dendrite of neocortical neurons.

1. Evidence for amplification of synaptic current by voltage-gated channels in dendrites of neocortical pyramidal neurons was demonstrated by examining the effect of specific channel blocking agents on the current arriving at the soma during iontophoresis of glutamate at a distal site on the apical dendrite. 2. Dendritic noninactivating Na+ channels were implicated in this voltage-dependent amplification of the transmitted current because it was maintained for > 1 s and because tetrodotoxin (TTX) eliminated much of this amplification. 3. Specific blockers of N-methyl-D-aspartate (NMDA) glutamate receptors reduced the amplitude of the glutamate-evoked current at all potentials and also reduced the non-TTX-sensitive component of voltage-dependent augmentation. The effects of TTX were identical whether or not NMDA channels were blocked. 4. We conclude that a persistent Na+ conductance exists in the apical dendrite of neocortical neurons. Together with the NMDA conductance at the synaptic site it provides a mechanism for the graded, voltage-dependent amplification of tonic, excitatory synaptic input. This amplification results in much more effective transmission of tonic excitatory current to the soma than would occur in a passive dendrite.