Ionization-induced electron trapping in ultrarelativistic plasma wakes.

The onset of trapping of electrons born inside a highly relativistic, 3D beam-driven plasma wake is investigated. Trapping occurs in the transition regions of a Li plasma confined by He gas. Li plasma electrons support the wake, and higher ionization potential He atoms are ionized as the beam is focused by Li ions and can be trapped. As the wake amplitude is increased, the onset of trapping is observed. Some electrons gain up to 7.6 GeV in a 30.5 cm plasma. The experimentally inferred trapping threshold is at a wake amplitude of 36 GV/m, in good agreement with an analytical model and PIC simulations.

Positron production by x rays emitted by betatron motion in a plasma wiggler.

Positrons in the energy range of 3-30 MeV, produced by x rays emitted by betatron motion in a plasma wiggler of 28.5 GeV electrons from the SLAC accelerator, have been measured. The extremely high-strength plasma wiggler is an ion column induced by the electron beam as it propagates through and ionizes dense lithium vapor. X rays in the range of 1-50 MeV in a forward cone angle of 0.1 mrad collide with a 1.7 mm thick tungsten target to produce electron-positron pairs. The positron spectra are found to be strongly influenced by the plasma density and length as well as the electron bunch length. By characterizing the beam propagation in the ion column these influences are quantified and result in excellent agreement between the measured and calculated positron spectra.

Hose instability and wake generation by an intense electron beam in a self-ionized gas.

The propagation of an intense relativistic electron beam through a gas that is self-ionized by the beam's space charge and wakefields is examined analytically and with 3D particle-in-cell simulations. Instability arises from the coupling between a beam and the offset plasma channel it creates when it is perturbed. The traditional electron hose instability in a preformed plasma is replaced with this slower growth instability depending on the radius of the ionization channel compared to the electron blowout radius. A new regime for hose stable plasma wakefield acceleration is suggested.

Multi-GeV energy gain in a plasma-wakefield accelerator.

A plasma-wakefield accelerator has accelerated particles by over 2.7 GeV in a 10 cm long plasma module. A 28.5 GeV electron beam with 1.8 x 10(10) electrons is compressed to 20 microm longitudinally and focused to a transverse spot size of 10 microm at the entrance of a 10 cm long column of lithium vapor with density 2.8 x 10(17) atoms/cm3. The electron bunch fully ionizes the lithium vapor to create a plasma and then expels the plasma electrons. These electrons return one-half plasma period later driving a large amplitude plasma wake that in turn accelerates particles in the back of the bunch by more than 2.7 GeV.

Plasma wakefield acceleration in self-ionized gas or plasmas.

Tunnel ionizing neutral gas with the self-field of a charged particle beam is explored as a possible way of creating plasma sources for a plasma wakefield accelerator [Bruhwiler et al., Phys. Plasmas (to be published)]. The optimal gas density for maximizing the plasma wakefield without preionized plasma is studied using the PIC simulation code OSIRIS [R. Hemker et al., in Proceeding of the Fifth IEEE Particle Accelerator Conference (IEEE, 1999), pp. 3672-3674]. To obtain wakefields comparable to the optimal preionized case, the gas density needs to be seven times higher than the plasma density in a typical preionized case. A physical explanation is given.

Cholinergic neurons of the dorsal pontine tegmentum projecting to the cerebellar vermal cortex of the kitten.

Although the whole cerebellar cortex receives cholinergic afferents, the source of origin of this projection has been clarified only for some corticocerebellar regions. Experiments were performed in kittens to investigate whether the two major cholinergic groups of the brainstem, the pedunculopontine (PPT) and laterodorsal tegmental nuclei (LDT), contribute to the cholinergic innervation of the cerebellar cortex, in particular the vermal cortex. Tegmento-cerebellar projecting neurons were identified by injecting the retrograde tracer rhodamine-labeled latex microspheres in the lobules V to VII of the cerebellar vermis. Subsequently, some of these tegmento-cerebellar neurons were demonstrated to be cholinergic by using the immunohistochemical technique for choline acetyltransferase (ChAT). Only a small portion of the ChAT-positive tegmental neurons projected to the cerebellar vermis. However, among the whole population of the retrogradely labeled tegmental neurons about one third were cholinergic. These cholinergic tegmento-cerebellar neurons were located in the PPT, LDT, and also within the locus coeruleus (LC) complex, where noradrenergic neurons predominate. Since the LC complex sends noradrenergic afferents to the cerebellar cortex, it appears that the dorsal pontine area contributes to the tegmento-cerebellar projections not only with noradrenergic but also with cholinergic afferents. The physiological significance of this cholinergic projection to the cerebellar cortex has been discussed.

The neurochemical and behavioral effects of beta-amyloid peptide(25-35).

Beta-amyloid protein (A beta) fragments have been shown to be neurotoxic and/or enhance neuronal vulnerability when injected into the hippocampus. We investigated alterations in monoamine contents, including norepinephrine (NE), 5-HT and dopamine (DA) in the rat locus coeruleus (LC) one week following the injection of beta-amyloid peptide fragment 25-35 (beta (25-35)) into the left dorsal hippocampal areas CA1-3. A single treatment of beta (25-35) had no effect on any monoamine levels. Rats that received two treatments (separated by 7 days) revealed significant elevations in NE, 5-HT, and 5-HIAA as compared with the control group injected with ddH2O. However, these changes were observed in the LC on the contralateral side, whereas the injected side exhibited no significant change. These effects may result from an enhanced synthesis of NE by the contralateral LC neurons to compensate for the loss of tyrosine hydroxylase and accompanying recurrent inhibition in a small number of their population. In a second experiment, the influence of beta (25-35) on spatial learning was evaluated using a Morris water maze task. Rats received bilateral injections of beta (25-35) into hippocampal areas CA1-3. The results indicate that beta (25-35)-treated rats exhibited significantly longer latencies and swim distances to locate the submerged platform than did members of the control group.

Neuropathology of synthetic beta-amyloid peptide analogs in vivo.

Considerable evidence exists demonstrating that beta-amyloid protein and its fragments 1-40 and 25-35 (beta (25-35)) are neurotoxic to cells in the rat hippocampus both in culture and in vivo. This neurotoxicity has been correlated to the aggregational state of the peptides. Previously we have shown that beta (25-35) produces a cavitational lesion in rat hippocampus and also reduces the enzyme or transmitter expressions in two subcortical structures whose axons project to the hippocampus: the locus coeruleus (LC) and the medial septum. In the present study, we further investigated the amino acid sequence that might be responsible for these effects. A series of synthetic peptide analogs of beta (25-35) with glycine substituted for serine, asparagine, lysine and methionine at positions 26, 27, 28 and 35, respectively, were injected at a 3 nmol dosage into the rat hippocampus once a week for 2 weeks. The damage to the hippocampus and immunohistochemistry of the LC and medial septum were examined 1 week following the second treatment. All of the synthetic peptides with glycine substitution produced damage to the hippocampal tissue. This damage was similar to that seen with beta (25-35). However, the reduction of enzyme expressions in the LC and medial septum was less from these substituted peptides than from that of beta (25-35). While beta (25-35) application resulted in a similar reduction of tyrosine hydroxylase (TH) and glutamate (Glu) immunoreactivities in the LC, only TH was significantly reduced in the substituted peptide groups. The least reduction of TH and Glu immunoreactivities in the LC was observed in rats treated with peptides in which glycine replaced either lysine or methionine. In the basal forebrain medial septum, the application of beta (25-35) resulted in a marked decrease in choline acetyltransferase (ChAT) immunoreactivity. This reduction was found to be less by each of the synthetic peptides. These results suggest that the biological activity of beta (25-35) is sensitive to changes in the primary structure of the peptide. Among the 4 amino acid residues examined, lysine and methionine at positions 28 and 35 appear to play more important roles in determining the action of beta (25-35).

Localization of glutamatergic neurons in the dorsolateral pontine tegmentum projecting to the spinal cord of the cat with a proposed role of glutamate on lumbar motoneuron activity.

Glutamate is considered to be a major excitatory neurotransmitter in the central nervous system. The presence of glutamate-like immunoreactive neurons in the rodent locus coeruleus has been reported previously. In this study we used both immunohistochemical and electrophysiological techniques to answer two major questions: (1) Is there any glutamate-like immunoreactivity in the catecholaminergic coeruleospinal system of the cat? (2) What is the physiological role, if any, of glutamate in descending locus coeruleus control of spinal motoneurons? Following injections of rhodamine-labeled latex microspheres or Fast Blue into the seventh lumbar segment of the spinal cord of the cat, retrogradely labeled cells were found throughout the rostrocaudal extent of the dorsolateral pontine tegmentum. They were primarily observed in the nucleus locus coeruleus and the Kolliker-Fuse nucleus. Some labeled cells were also present in the nucleus subcoeruleus and, to a lesser extent, in the parabrachial nuclei. Data from immunohistochemical studies indicate that 86% of all dorsolateral pontine tegmentum neurons that project to the spinal cord contain glutamate-like immunoreactivity, and 77% co-contain both glutamate- and tyrosine hydroxylase-like immunoreactivity. Electrical stimulation (four pulses of 500 microseconds duration at 500 Hz; intensity = 50-200 microA) of the locus coeruleus, in decerebrate cats, consistently induced lumbar motoneuron discharges recordable ipsilaterally as ventral root responses. These motoneuronal responses were reversibly antagonized following chemical inactivation of noradrenergic locus coeruleus neurons by local infusion of the alpha 2-adrenergic agonist clonidine, suggesting the locus coeruleus neurons to be the main source of evoked ventral root responses. Additionally, the evoked ventral root responses were reversibly reduced by 34.20 +/- 4.45% (mean +/- S.E.M.) upon intraspinal injections of the non-N-methyl-D-aspartate receptor antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione, into the ventral horn of seventh lumbar spinal cord segment (three to four injections, 20 nmol in 0.2 microliter of 0.1 M Tris-buffered saline for each injection). Similar volumes of vehicle injections had no significant effect on the locus coeruleus-evoked ventral root responses. These ventral root responses were also partially blocked (62.30 +/- 11.76%) by intravenous administration of the alpha 1-adrenergic receptor antagonist prazosin (20 micrograms/kg). In the light of several anatomical reports of noradrenergic and glutamatergic terminals in close contact with spinal motoneurons, our present findings suggest that the locus coeruleus-evoked ventral root response probably involves the synaptic release of both norepinephrine and glutamate onto lumbar motoneurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Anatomical evidence for the presence of glutamate or enkephalin in noradrenergic projection neurons of the locus coeruleus.

This paper reviews the anatomical evidence for the presence of glutamate (GLU) in noradrenergic neurons of the nucleus locus coeruleus (LC) and adjacent nuclei in the dorsolateral pontine tegmentum (DLPT) that project to the spinal cord, cerebellum, or cerebral cortex. Additionally, the evidence for the existence of methionine-enkephalin (ENK) in noradrenergic neurons of the DLPT that project to the spinal cord of the cat is reviewed. In these studies, we have combined the retrograde transport of either Fast Blue (FB), rhodamine labeled latex microspheres (MS), or rhodamine labeled dextran and indirect immunofluorescence histochemistry to determine whether the neurons that contain tyrosine hydroxylase (TH) and project to these terminal fields also contain GLU or ENK. The neurons of the cat that project to the spinal cord, cerebellum, and neocortex were observed in the nucleus LC and Kölliker-Fuse (KF) nucleus. They were also present, to a lesser extent, in the nucleus subcoeruleus (SC) and nuclei parabrachialis medialis (PBM) and lateralis (PBL). In the rat the majority of the neurons that project to the neocortex and hippocampus were located in the nucleus LC. Our data revealed a major proportion of these neurons to be immunostained for both GLU and TH (cat, rat), or ENK and TH (cat). Functional implications of such colocalized neurochemicals within individual LC projection neurons are discussed.

Bulbospinal neurons of the cat that co-contain serotonin and methionine enkephalin.

The present study utilizes a combined retrograde transport of Fast Blue (or rhodamine-labeled latex microspheres) and simultaneous immunofluorescence technique to demonstrate directly the coexistence of serotonin and methionine enkephalin in bulbospinal neurons of the cat. The bulbospinal neurons that immunostained for both serotonin and enkephalin were observed, without any distinct somatotopic organization, in the nuclei raphe pallidus, obscurus and magnus. They were also observed in the nucleus reticularis magnocellularis and the ventrolateral medulla (cell group B1/3). Among the bulbospinal neurons encountered within individual 5-HT-rich medullary nuclei, high proportions of these neurons co-containing serotonin and methionine enkephalin were evidenced in the nucleus raphe obscurus (64%) and nucleus raphe pallidus (56%), less so in cell group B1/3 (41%), nucleus raphe magnus (39%), and the nucleus reticularis magnocellularis (29%). Physiological significance of such a morphological substrate is discussed.

Cotransmitter-mediated locus coeruleus action on motoneurons.

This article reviews evidence for a direct noradrenergic projection from the dorsolateral pontine tegmentum (DLPT) to spinal motoneurons. The existence of this direct pathway was first inferred by the observation that antidromically evoked responses occur in single cells in the locus coeruleus (LC), a region within the DLPT, following electrical stimulation of the ventral horn of the lumbar spinal cord of the cat. We subsequently confirmed that there is a direct noradrenergic pathway from the LC and adjacent regions of the DLPT to the lumbar ventral horn using anatomical studies that combined retrograde tracing with immunohistochemical identification of neurotransmitters. These anatomical studies further revealed that many of the noradrenergic neurons in the LC and adjacent regions of the DLPT of the cat that send projections to the spinal cord ventral horn also contain colocalized glutamate (Glu) or enkephalin (ENK). Recent studies from our laboratory suggest that Glu and ENK may function as cotransmitters with norepinephrine (NE) in the descending pathway from the DLPT. Electrical stimulation of the LC evokes a depolarizing response in spinal motoneurons that is only partially blocked by alpha 1 adrenergic antagonists. In addition, NE mimicks only the slowly developing and not the fast component of LC-evoked depolarization. Furthermore, the depolarization evoked by LC stimulation is accompanied by a decrease in membrane resistance, whereas that evoked by NE is accompanied by an increased resistance. That Glu may be a second neurotransmitter involved in LC excitation of motoneurons is supported by our observation that the excitatory response evoked in spinal cord ventral roots by electrical stimulation of the LC is attenuated by a non-N-methyl-D-aspartate glutamatergic antagonist. ENK may participate as a cotransmitter with NE to mediate LC effects on lumbar monosynaptic reflex (MSR) amplitude. Electrical stimulation of the LC has a biphasic effect on MSR amplitude, facilitation followed by inhibition. Adrenergic antagonists block only the facilitator effect of LC stimulation on MSR amplitude, whereas the ENK antagonist naloxone reverses the inhibition. The chemical heterogeneity of the cat DLPT system and the differential responses of motoneurons to the individual cotransmitters help to explain the diversity of postsynaptic potentials that occur following LC stimuli.

Hippocampal beta-amyloid reduces locus coeruleus glutamate and tyrosine hydroxylase.

The effects of intrahippocampally injected beta-amyloid protein (beta-AP) on glutamate- (Glu) and tyrosine hydroxylase (TH)-like immunoreactivities in the neurons of the locus coeruleus (LC) were studied in rats. A synthetic peptide or the vehicle alone was injected into the hippocampus as controls. All injections were made once a week (two or three injections; 3 nmol in 2 microliters of distilled water). Fluorescent microspheres (either alone or with one of the peptides) were also injected into the hippocampus to identify coeruleo-hippocampal neurons. The results revealed cell loss in the hippocampus at the site near beta-AP or control peptide deposition. Furthermore, in beta-AP/microsphere injected animals, only 22.4% and 49.6% of hippocampal projection neurons contained Glu and TH, respectively, compared to 88.4% and 85.3% in the animals that received control peptide with microspheres. Our results suggest that beta-AP has an effect on noradrenergic cells whose axons project to the hippocampus. These effects may contribute to the TH cell loss in the LC of Alzheimer's brains.

Existence of glutamate in noradrenergic locus coeruleus neurons of rodents.

This study distinguished three types of immunolabeled neurons in nucleus locus coeruleus (LC) of the rat and mouse: cells single labeled either for tyrosine hydroxylase-like immunoreactivity (TH-LI) or glutamate (Glu)-LI, and those double labeled for both antigens. Although the double labeled neurons tend to be located in the middle and ventral thirds of the rat LC nucleus, throughout its rostrocaudal extent, such feature was not apparent in the mouse. Quantitatively a majority of neurons cocontaining TH- and Glu-LI were commonly observed in the rat (62%) and mouse (77%) LC. Additional studies utilizing the combined retrograde and immunohistochemical labeling revealed that such a high incidence of coexistence of the TH- and Glu-LI was also represented by coeruleocortical neurons in the rat (69% and 75% of all ipsilateral and contralateral projection cells, respectively). A possible role of coeruleocortical neurons involvement in Glu- and norepinephrine-mediated target neuron dysfunction is discussed.

Immunohistochemical localization of glutamate-containing neurons in the lateral reticular nucleus projecting to the cerebellar vermis in the kitten.

The lateral reticular nucleus (LRN) and afferents to the cerebellum are known to contain glutamate-like immunoreactive (Glu-LI) neurons and axons, respectively. However, such a direct link between the Glu-LI LRN neurons and the cerebellar vermal cortex has not been identified. In this study we have combined the retrograde transport of rhodamine labeled latex microspheres and immunofluorescence histochemistry to determine the locations of Glu-LI neurons of the kitten reticulocerebellar system. Following microsphere injections into the cerebellar vermis (lobules V-VII), retrogradely labeled neurons were encountered throughout the rostrocaudal extent of the LRN. More than 60% (n = 3 kittens) of these retrogradely labeled neurons were immunostained for Glu-like immunoreactivity. Our observations of the Glu-like immunoreactivity in a majority of the reticulocerebellar neurons suggest that Glu in these neurons may participate in LRN's control of target neuron activities in the cerebellar vermis of kittens.

Opioid action on spinal cord reflexes due to dorsolateral pontine tegmentum stimulation.

Electrical stimulation of the dorsolateral pontine tegmentum (DLPT) produces phasic facilitatory and inhibitory actions on the lumbar spinal monosynaptic reflexes (MSRs) of both flexor and extensor muscle nerves in the decerebrate cat. Naloxone, an opioid receptor antagonist, given intravenously or intraspinally enhanced the DLPT-induced potentiation of MSRs in most of the reflexes studied. However, systemic naloxone had no significant effect on the unconditioned MSR of the spinal cord. Intraspinal microinjections of naloxone significantly attenuated the DLPT-induced inhibition of MSRs of both flexors and extensors, similar to the action of systemic injection of naloxone, indicating a direct opioid action at the spinal ventral horn level upon DLPT stimulation. Results of the present experiment further support the anatomical finding that there are pontospinal enkephalinergic pathways in the cat, and indicate that these descending pathways modulate spinal motor outflow.

Application of compiled BASIC in developing software for collection and analysis of neuronal firing frequency data.

Two programs are described which use an IBM-AT compatible personal computer, equipped with an analog-to-digital converter, to collect and analyze electrophysiological data. The first program is used to determine neuron firing rates during intracellular experiments and to save these results on disk. The second program is used to perform off-line analysis of the frequency data. Both programs are written entirely in the well known BASIC language and the Microsoft QuickBASIC compiler is employed for their use. All of the necessary hardware can be purchased commercially. In this paper emphasis is placed on the strategies and limitations involved when this high-level language is applied to tasks often needed in data acquisition and analysis. Both on- and off-line collection schemes are considered. This software is available from the authors.

Immunohistochemical evidence for coexistence of methionine-enkephalin and tyrosine hydroxylase in neurons of the locus coeruleus complex projecting to the spinal cord of the cat.

Previous studies have revealed the presence of pontospinal neurons with either methionine-enkephalin- or tyrosine hydroxylase-like immunoreactivity in the dorsolateral pontine tegmentum of the cat. Using a combined fast blue retrograde transport technique and simultaneous immunofluorescence histochemistry, the present study was designed to reveal the coexistence of enkephalin and tyrosine hydroxylase in cat coerulospinal neurons and to determine if and to what extent the coerulospinal pathway is heterogeneous. Fast blue-labelled neurons with tyrosine hydroxylase- and enkephalin-like immunoreactivities were found in the nucleus locus coeruleus, nucleus subcoeruleus, Kölliker-Fuse nucleus, and the medial and lateral parabrachial nuclei. Approximately 87% of tyrosine hydroxylase-like immunoreactive neurons had enkephalin-like immunoreactivity, whereas about 76% of the enkephalin-like immunoreactive neurons had tyrosine hydroxylase-like immunoreactivity. About 71% of all coerulospinal neurons exhibited both tyrosine hydroxylase- and enkephalin-like immunoreactivities. These findings indicate that coerulospinal activity may lead to spinal cord effects reflecting both norepinephrine and enkephalin activity in most cases but do not rule out each transmitter's isolated functions.

Effects of temperature and analyte application technique on neuron-based chemical sensing.

Results are presented which enhance the field of neuron-based sensing by providing insight on the effects of operating temperature and analyte application technique (pulse versus back-mixed) on sensing properties. In these studies, serotonin sensing attributes of giant visceral neurons VV1 and VV2 from the pond snail Lymnea stagnalis were measured. Experiments using a rapid fluid-exchange system reveal a concentration-dependent increase in maximum firing frequency similar to that reported earlier for a slow well-mixed application. With a rapid application, however, the maximum firing frequency is reached more quickly, and there is less cell-to-cell variability in both the maximum response and sensitivity. Given an application technique, an increase in temperature causes an increase in sensitivity and maximum firing frequency, as well as a decrease in the time required for the response to return to baseline following removal of the analyte. To provide insights on the kinetics of the serotonin-induced response, the effects of temperature and concentration on the rates of activation, recovery and desensitization were examined in detail. In general, it was found that an increase in temperature increases the rates of activation and desensitization, while the effects on recovery were not apparent. In addition, both the rates of activation and desensitization have a direct dependence on concentration while the rate of recovery has an inverse dependence.

Optimization of glass microelectrode properties by response surface methodology.

Glass microelectrodes filled with electrolyte solutions are standard tools for electrophysiological studies. However, for any given application, there are limitations to the properties of the microelectrode, such as impedance and shank length, that can yield satisfactory results. The trial and error approach in pulling electrodes with the desired properties can be time consuming. The use of a response surface procedure which allows the experimenter to change more than one factor at a time and therefore determine the desired puller condition more efficiently is demonstrated. Also, design improvements for the World Precision Instrument, Model PUL-1, Microelectrode puller, used in this study are suggested.