bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid, Activation-Induced Redistribution of Ionotropic Glutamate Receptors in Cultured Hippocampal Neurons
Authors: Lissin Dmitri V. , Carroll Reed C. , Nicoll Roger A. , Malenka Robert C. , Zastrow Mark von
Published: April 2018

We have examined the membrane localization of an AMPA receptor subunit (GluR1) and an NMDA receptor subunit (NR1) endogenously expressed in primary cultures of rat hippocampal neurons. In unstimulated cultures, both GluR1 and NR1 subunits were concentrated in SV2-positive synaptic clusters associated with dendritic shafts and spines. Within 5 min after the addition of 100 μmglutamate to the culture medium, a rapid and selective redistribution of GluR1 subunits away from a subset of synaptic sites was observed. This redistribution of GluR1 subunits was also induced by AMPA, did not require NMDA receptor activation, did not result from ligand-induced neurotoxicity, and was reversible after the removal of agonist. The activation-induced redistribution of GluR1 subunits was associated with a pronounced (∼50%) decrease in the frequency of miniature EPSCs, consistent with a role of GluR1 subunit redistribution in mediating rapid regulation of synaptic efficacy. We conclude that ionotropic glutamate receptors are regulated in native neurons by rapid, subtype-specific membrane trafficking, which may modulate synaptic transmission in response to physiological or pathophysiological activation.
Sustained and Accelerating Activity at Two Discrete Sites Generate Epileptiform Discharges in Slices of Piriform Cortex
Authors: Demir Rezan , Haberly Lewis B. , Jackson Meyer B.
Published: April 2018

When near-threshold electrical stimulation is used to evoke epileptiform discharges in brain slices, a latent period of up to 150 msec elapses before the discharge begins. During this period most neurons are silent, and abnormal electrical activity is difficult to detect with microelectrodes. A fundamental question about epileptiform activity concerns how synchronous discharges arise abruptly in a relatively quiescent slice. This issue was addressed here by using voltage imaging techniques to study epileptiform discharges in rat piriform cortex slices. These experiments revealed two distinct forms of electrical activity during the latent period. (1) A steeply increasing depolarization, referred to here as onset activity, has been described previously and occurs at the site of discharge onset. (2) A sustained depolarization that precedes onset activity, referred to here as plateau activity, has not been described previously. Plateau and onset activity occurred in different subregions of the endopiriform nucleus (a region of high seizure susceptibility). When cobalt or kynurenic acid was applied focally to inhibit electrical activity at the site of plateau activity, discharges were blocked. However, application of these agents to other nearby sites (except the site of onset) failed to block discharges. Plateau activity represents a novel form of electrical activity that precedes and is necessary for epileptiform discharges. Discharges thus are generated in a sequential process by two spatially distinct neuronal circuits. The first circuit amplifies and sustains activity initiated by the stimulus, and the second generates the actual discharge in response to an excitatory drive from the first.
Suppression of Postischemic Hippocampal Nerve Growth Factor Expression by a c-fosAntisense Oligodeoxynucleotide
Authors: Cui Jian-Kun , Hsu Chung Y. , Liu Philip K.
Published: April 2018

We examined the uptake and distribution of an antisense phosphorothioated oligodeoxynucleotide (s-ODN) to c-fos, rncfosr115, infused into the left cerebral ventricle of male Long–Evans rats and the effect of this s-ODN on subsequent Fos, NGF, neurotrophin-3 (NT-3), and actin expression. To establish the uptake and turnover of s-ODN in the brain, we studied the copurification of the immunoreactivity of biotin with biotinylated s-ODN that was recovered from different regions of the brain. A time-dependent diffusion and the localization of s-ODN were further demonstrated by labeling the 3′-OH terminus of s-ODNin situwith digoxigenin-dUTP using terminal transferase and detection using anti-digoxigenin IgG–FITC. Cellular uptake of the s-ODN was evident in both the hippocampal and cortical regions, consistent with a gradient originating at the ventricular surface. Degradation of the s-ODN was observed beginning 48 hr after delivery. The effectiveness of c-fosantisense s-ODN was demonstrated by its suppression of postischemic Fos expression, which was accompanied by an inhibition of ischemia-induced NGF mRNA expression in the dentate gyrus. Infusion of saline, the sense s-ODN, or a mismatch antisense s-ODN did not suppress Fos expression. That this effect of c-fosantisense s-ODN was specific to NGF was demonstrated by its lack of effect on the postischemic expression of the NT-3 and β-actin genes. Our results demonstrate that c-fosantisense s-ODN blocks selected downstream events and support the contention that postischemic Fos regulates the subsequent expression of the NGF gene and that Fos expression may have a functional component in neuroregeneration after focal cerebral ischemia-reperfusion.
Odor-Induced, Activity-Dependent Transneuronal Gene InductionIn Vitro: Mediation by NMDA Receptors
Authors: Puche A. C. , Shipley M. T.
Published: April 2018

Expression of tyrosine hydroxylase (TH) by juxtaglomerular (JG) neurons of the olfactory bulb (OB) requires innervation of the bulb by olfactory receptor neurons (ORNs). ORN lesion selectively downregulates TH in JG neurons. In reversible odor deprivation, TH expression is downregulated as the naris is closed and then upregulated upon naris reopening. The mechanism or mechanisms regulating this dependence are unknown. TH expression could be regulated by trophic factor release and/or synaptic activity from ORN terminals. We investigated TH expression in cocultures of dissociated postnatal rat OB cells and embryonic olfactory neuroepithelium (OE) slice explants. TH-positive neurons in control dissociated OB cell cultures alone comprise only a small fraction of the total population of cells present in the culture. However, when OE slice explants are cocultured with dispersed OB cells, there is a mean 2.4-fold increase in the number of TH-positive neurons. ORNsin vivouse glutamate as a neurotransmitter. Broad spectrum excitatory amino acid antagonists (kyurenic acid) or selective antagonists of the NMDA receptor (APV) both prevent induction of TH expression in OE–OB cocultures. Furthermore, pulse application of NMDA stimulates TH expression in OB neurons in the absence of OE.In vitro, OB TH neurons express NMDA receptors, suggesting that NMDA stimulation is acting directly on TH neurons. Exposure of OE explants to natural odorants results in upregulation of TH, presumably through increased ORN activity, which could be blocked by APV. These findings indicate that odorant-stimulated glutamate release by ORN terminals regulates TH expression via NMDA receptors on JG dopaminergic neurons.
Stimulation of Myelin Basic Protein Gene Transcription by Fyn Tyrosine Kinase for Myelination
Authors: Umemori Hisashi , Kadowaki Yasunori , Hirosawa Kazushige , Yoshida Yutaka , Hironaka Katsunori , Okano Hideyuki , Yamamoto Tadashi
Published: April 2018

Myelin is synthesized about the time of birth. The Src-family tyrosine kinase Fyn is involved in the initial events of myelination. Fyn is present in myelin-forming cells and is activated through stimulation of cell surface receptors such as large myelin-associated glycoprotein (L-MAG). Here we show that Fyn stimulates transcription of the myelin basic protein (MBP) gene for myelination. MBP is a major component of the myelin membrane. In 4-week-old Fyn-deficient mice, MBP is significantly reduced, and electron microscopic analysis showed that myelination is delayed, compared with wild-type mice. The Fyn-deficient mice had thinner, more irregular myelin than the wild-type. We found that Fyn stimulates the promoter activity of the MBP gene by approximately sevenfold. The region responsible for the transactivation by Fyn is located between nucleotides −675 and −647 with respect to the transcription start site. Proteins binding to this region were found by gel shift study, and the binding activity correlates with Fyn activity during myelination. These results suggest that transactivation of the MBP gene by Fyn is important for myelination.
Dopamine Selectively Inhibits the Direct Cortical Pathway to the CA1 Hippocampal Region
Authors: Otmakhova Nonna A. , Lisman John E.
Published: April 2018

The perforant path input (pp) is a major direct source of specific sensory information for the CA1 hippocampal region. The termination area of this pathway, the stratum lacunosum-moleculare, has the highest concentration of dopamine receptors in the hippocampus. We have examined the properties of the pp input and its modulation by dopamine. The input is glutamatergic and has a larger NMDA component than the Schaffer collateral (sc) input. Dopamine strongly inhibits the response to pp stimulation (IC50∼3 μm) but not the response to sc stimulation. Dopamine reduces both the NMDA and AMPA components of transmission at the pp and increases paired-pulse facilitation. In the sc, the NMDA component but not the AMPA component is decreased, and paired-pulse facilitation is not affected. The effect of dopamine on the pp does not depend on GABAAinhibition but is reduced by the antagonists of both D1 and D2 families of dopamine receptors. The effect is not completely blocked by the combination of D1 and D2 antagonists, but is completely blocked by the atypical neuroleptic clozapine. Our results provide the first evidence for strong dopaminergic control of transmission in the perforant path. By inhibiting this pathway, dopamine hyperfunction and/or NMDA hypofunction abnormalities implicated in schizophrenia may isolate CA1 from its main source of sensory information.
The Organization of Cerebellar and Basal Ganglia Outputs to Primary Motor Cortex as Revealed by Retrograde Transneuronal Transport of Herpes Simplex Virus Type 1
Authors: Hoover John E. , Strick Peter L.
Published: April 2018

We used retrograde transneuronal transport of herpes simplex virus type 1 to map the origin of cerebellar and basal ganglia “projections” to leg, arm, and face areas of the primary motor cortex (M1). Four to five days after virus injections into M1, we observed many densely labeled neurons in localized regions of the output nuclei of the cerebellum and basal ganglia. The largest numbers of these neurons were found in portions of the dentate nucleus and the internal segment of the globus pallidus (GPi). Smaller numbers of labeled neurons were found in portions of the interpositus nucleus and the substantia nigra pars reticulata. The distribution of neuronal labeling varied with the cortical injection site. For example, within the dentate, neurons labeled from leg M1 were located rostrally, those from face M1 caudally, and those from arm M1 at intermediate levels. In each instance, labeled neurons were confined to approximately the dorsal third of the nucleus. Within GPi, neurons labeled from leg M1 were located in dorsal and medial regions, those from face M1 in ventral and lateral regions, and those from arm M1 in intermediate regions. These results demonstrate that M1 is the target of somatotopically organized outputs from both the cerebellum and basal ganglia. Surprisingly, the projections to M1 originate from only 30% of the volume of the dentate and <15% of GPi. Thus, the majority of the outputs from the cerebellum and basal ganglia are directed to cortical areas other than M1.
Active Motor Neurons Potentiate Their Own Sensory Inputs via Glutamate-Induced Long-Term Potentiation
Authors: Le Ray Didier , Cattaert Daniel
Published: April 2018

Adaptive motor control is based mainly on the processing and integration of proprioceptive feedback information. In crayfish walking leg, many of these operations are performed directly by the motor neurons (MNs), which are connected monosynaptically by sensory afferents (CBTs) originating from a chordotonal organ that encodes vertical limb movements. Anin vitropreparation of the crayfish CNS was used to investigate a new control mechanism exerted directly by motor neurons on the sensory inputs themselves. Paired intracellular recordings demonstrated that, in the absence of any presynaptic sensory firing, the spiking activity of a leg MN is able long-lastingly to enhance the efficacy of the CBT–MN synapses. Moreover, this effect is specific to the activated MN because no changes were induced at the afferent synapses of a neighboring silent MN. We report evidence that long-term potentiation (LTP) of the monosynaptic EPSP involves a retrograde system of glutamate transmission from the postsynaptic MN, which induces the activation of a metabotropic glutamate receptor located presynaptically on the CBTs. We demonstrate that LTP at crayfish sensory–motor synapses results exclusively from the long-lasting enhancement of release of acetylcholine from presynaptic sensory afferent terminals, without inducing any modifications in postsynaptic MN properties. Our data indicate that this positive feedback control represents a functional mechanism that may play a key role in the auto-organization of sensory–motor networks.
Dopamine Quinone Formation and Protein Modification Associated with the Striatal Neurotoxicity of Methamphetamine: Evidence against a Role for Extracellular Dopamine
Authors: LaVoie Matthew J. , Hastings Teresa G.
Published: April 2018

Methamphetamine-induced toxicity has been shown to require striatal dopamine and to involve mechanisms associated with oxidative stress. Dopamine is a reactive molecule that can oxidize to form free radicals and reactive quinones. Although this has been suggested to contribute to the mechanism of toxicity, the oxidation of dopamine has never been directly measured after methamphetamine exposure. In this study we sought to determine whether methamphetamine-induced toxicity is associated with the oxidation of dopamine by measuring the binding of dopamine quinones to cysteinyl residues on protein. We observed that administration of neurotoxic doses of methamphetamine to rats resulted in a two- to threefold increase in protein cysteinyl-dopamine in the striatum 2, 4, and 8 hr after treatment. When methamphetamine was administered at an ambient temperature of 5°C, no increase in dopamine oxidation products was observed, and toxicity was prevented. Furthermore, as shown by striatal microdialysis, animals treated with methamphetamine at 5°C showed DA release identical to that of animals treated at room temperature. These data suggest that the toxicity of methamphetamine and the associated increase in dopamine oxidation are not exclusively the result of increases in extracellular dopamine. Because dopamine-induced modifications of protein structure and function may result in cellular toxicity, it is likely that dopamine oxidation contributes to methamphetamine-induced toxicity to dopamine terminals, adding support to the role of dopamine and the evidence of oxidative stress in this lesion model.
Neural Coding of Sound Frequency by Cricket Auditory Receptors
Authors: Imaizumi Kazuo , Pollack Gerald S.
Published: April 2018

Crickets provide a useful model to study neural processing of sound frequency. Sound frequency is one parameter that crickets use to discriminate between conspecific signals and sounds made by predators, yet little is known about how frequency is represented at the level of auditory receptors. In this paper, we study the physiological properties of auditory receptor fibers (ARFs) by making single-unit recordings in the cricketTeleogryllus oceanicus. Characteristic frequencies (CFs) of ARFs are distributed discontinuously throughout the range of frequencies that we investigated (2–40 kHz) and appear to be clustered around three frequency ranges (≤5.5, 10–12, and ≥18 kHz). A striking characteristic of cricket ARFs is the occurrence of additional sensitivity peaks at frequencies other than CFs. These additional sensitivity peaks allow crickets to detect sound over a wide frequency range, although the CFs of ARFs cover only the frequency bands mentioned above. To the best of our knowledge, this is the first example of the extension of an animal’s hearing range through multiple sensitivity peaks of auditory receptors.
Responses to the Sensory Properties of Fat of Neurons in the Primate Orbitofrontal Cortex
Authors: Rolls Edmund T. , Critchley Hugo D. , Browning Andrew S. , Hernadi Istvan , Lenard Laszlo
Published: April 2018

The primate orbitofrontal cortex is a site of convergence of information from primary taste, olfactory, and somatosensory cortical areas. We describe the responses of a population of single neurons in the orbitofrontal cortex that responds to fat in the mouth. The neurons respond, when fatty foods are being eaten, to pure fat such as glyceryl trioleate and also to substances with a similar texture but different chemical composition such as paraffin oil (hydrocarbon) and silicone oil [Si(CH3)2O)n]. This is evidence that the neurons respond to the oral texture of fat, sensed by the somatosensory system. Some of the population of neurons respond unimodally to the texture of fat. Other single neurons show convergence of taste inputs, and others of olfactory inputs, onto single neurons that respond to fat. For example, neurons were found that responded to the mouth feel of fat and the taste of monosodium glutamate (both found in milk), or to the mouth feel of fat and to odor. Feeding to satiety reduces the responses of these neurons to the fatty food eaten, but the neurons still respond to some other foods that have not been fed to satiety. Thus sensory-specific satiety for fat is represented in the responses of single neurons in the primate orbitofrontal cortex.

Fat is an important constituent of food that affects its palatability and nutritional effects. The findings described provide evidence that the reward value (or pleasantness) of the mouth feel of fat is represented in the primate orbitofrontal cortex and that the representation is relevant to appetite.
Increased Methamphetamine Neurotoxicity in Heterozygous Vesicular Monoamine Transporter 2 Knock-Out Mice
Authors: Fumagalli Fabio , Gainetdinov Raul R. , Wang Yan-Min , Valenzano Kenneth J. , Miller Gary W. , Caron Marc G.
Published: April 2018

Methamphetamine (METH) is a powerful psychostimulant that is increasingly abused worldwide. Although it is commonly accepted that the dopaminergic system and oxidation of dopamine (DA) play pivotal roles in the neurotoxicity produced by this phenylethylamine, the primary source of DA responsible for this effect has remained elusive. In this study, we used mice heterozygous for vesicular monoamine transporter 2 (VMAT2 +/− mice) to determine whether impaired vesicular function alters the effects of METH. METH-induced dopaminergic neurotoxicity was increased in striatum of VMAT2 +/− mice compared with wild-type mice as revealed by a more consistent DA and metabolite depletion and a greater decrease in dopamine transporter expression. Interestingly, increased METH neurotoxicity in VMAT2 +/− mice was accompanied by less pronounced increase in extracellular DA and indices of free radical formation compared with wild-type mice. These results indicate that disruption of vesicular monoamine transport potentiates METH-induced neurotoxicityin vivoand point, albeit indirectly, to a greater contribution of intraneuronal DA redistribution rather than extraneuronal overflow on mediating this effect.
Selective Effects ofneuronal-synaptobrevinMutations on Transmitter Release Evoked by Sustained Versus Transient Ca2+Increases and by cAMP
Authors: Yoshihara Motojiro , Ueda Atsushi , Zhang Dawei , Deitcher David L. , Schwarz Thomas L. , Kidokoro Yoshiaki
Published: April 2018

Synaptobrevin is a key constituent of the synaptic vesicle membrane. Theneuronal-synaptobrevin(n-syb) gene inDrosophilais essential for nerve-evoked synaptic currents, but miniature excitatory synaptic currents (mESCs) remain even in the complete absence of this gene. To further characterize the defect in these mutants, we have examined conditions that stimulate secretion. Despite the inability of an action potential to trigger fusion, high K+saline could increase the frequency of mESCs 4- to 17-fold in a Ca2+-dependent manner, and the rate of fusion approached 25% of that seen in wild-type synapses under the same conditions. Similarly, the mESC frequency inn-sybnull mutants could be increased by a Ca2+ionophore, A23187, and by black widow spider venom. Thus, the ability of the vesicles to fuse in response to sustained increases in cytosolic Ca2+persisted in the absence of this protein. Tetanic stimulation could also increase the frequency of mESCs, particularly toward the end of a train and after the train of stimuli. In contrast, these mutants did not respond to an elevation of cAMP induced by an activator of adenylyl cyclase, forskolin, or a membrane-permeable analog of cAMP, dibutyryl cAMP, which in wild-type synapses causes a marked increase in the mESC frequency even in the absence of external Ca2+. These results are discussed in the context of models that invoke a special role for n-syb in coupling fusion to the transient, local changes in Ca2+and an as yet unidentified target of cAMP.
Chlorpromazine Inhibits Miniature GABAergic Currents by Reducing the Binding and by Increasing the Unbinding Rate of GABAAReceptors
Authors: Mozrzymas Jerzy W. , Barberis Andrea , Michalak Krystyna , Cherubini Enrico
Published: April 2018

Recent studies have emphasized that nonequilibrium conditions of postsynaptic GABAAreceptor (GABAAR) activation is a key factor in shaping the time course of IPSCs (Puia et al., 1994; Jones and Westbrook, 1995). Such nonequilibrium, resulting from extremely fast agonist time course, may affect the interaction between pharmacological agents and postsynaptic GABAARs. In the present study we found that chlorpromazine (CPZ), a widely used antipsychotic drug known to interfere with several ligand and voltage-gated channels, reduces the amplitude and accelerates the decay of miniature IPSCs (mIPSCs). A good qualitative reproduction of the effects of CPZ on mIPSCs was obtained when mIPSCs were mimicked by responses to ultrafast GABA applications to excised patches. Our experimental data and model simulations indicate that CPZ affects mIPSCs by decreasing the binding (kon) and by increasing the unbinding (koff) rates of GABAARs. Because of reduction ofkonby CPZ, the binding reaction becomes rate-limiting, and agonist exposure of GABAARs during mIPSC is too short to activate the receptors to the same extent as in control conditions. The increase in unbinding rate is implicated as the mechanism underlying the acceleration of mIPSC decaying phase. The effect of CPZ on GABAAR binding rate, resulting in slower onset of GABA-evoked currents, provides a tool to estimate the speed of synaptic clearance of GABA. Moreover, the onset kinetics of recorded responses allowed the estimate the peak synaptic GABA concentration.
Adenylyl Cyclase Activation Modulates Activity-Dependent Changes in Synaptic Strength and Ca2+/Calmodulin-Dependent Kinase II Autophosphorylation
Authors: Makhinson Michael , Chotiner Jennifer K. , Watson Joseph B. , O’Dell Thomas J.
Published: April 2018

Activation of the Ca2+- and calmodulin-dependent protein kinase II (CaMKII) and its conversion into a persistently activated form by autophosphorylation are thought to be crucial events underlying the induction of long-term potentiation (LTP) by increases in postsynaptic Ca2+. Because increases in Ca2+can also activate protein phosphatases that oppose persistent CaMKII activation, LTP induction may also require activation of signaling pathways that suppress protein phosphatase activation. Because the adenylyl cyclase (AC)–protein kinase A signaling pathway may provide a mechanism for suppressing protein phosphatase activation, we investigated the effects of AC activators on activity-dependent changes in synaptic strength and on levels of autophosphorylated αCaMKII (Thr286). In the CA1 region of hippocampal slices, briefly elevating extracellular Ca2+induced an activity-dependent, transient potentiation of synaptic transmission that could be converted into a persistent potentiation by the addition of phosphatase inhibitors or AC activators. To examine activity-dependent changes in αCaMKII autophosphorylation, we replaced electrical presynaptic fiber stimulation with an increase in extracellular K+to achieve a more global synaptic activation during perfusion of high Ca2+solutions. In the presence of the AC activator forskolin or the protein phosphatase inhibitor calyculin A, this treatment induced a LTP-like synaptic potentiation and a persistent increase in autophosphorylated αCaMKII levels. In the absence of forskolin or calyculin A, it had no lasting effect on synaptic strength and induced a persistent decrease in autophosphorylated αCaMKII levels. Our results suggest that AC activation facilitates LTP induction by suppressing protein phosphatases and enabling a persistent increase in the levels of autophosphorylated CaMKII.
Nuclear and Neuropil Aggregates in Huntington’s Disease: Relationship to Neuropathology
Authors: Gutekunst Claire-Anne , Li Shi-Hua , Yi Hong , Mulroy James S. , Kuemmerle Stefan , Jones Randi , Rye David , Ferrante Robert J. , Hersch Steven M. , Li Xiao-Jiang
Published: April 2018

The data we report in this study concern the types, location, numbers, forms, and composition of microscopic huntingtin aggregates in brain tissues from humans with different grades of Huntington’s disease (HD). We have developed a fusion protein antibody against the first 256 amino acids that preferentially recognizes aggregated huntingtin and labels many more aggregates in neuronal nuclei, perikarya, and processes in human brain than have been described previously. Using this antibody and human brain tissue ranging from presymptomatic to grade 4, we have compared the numbers and locations of nuclear and neuropil aggregates with the known patterns of neuronal death in HD. We show that neuropil aggregates are much more common than nuclear aggregates and can be present in large numbers before the onset of clinical symptoms. There are also many more aggregates in cortex than in striatum, where they are actually uncommon. Although the striatum is the most affected region in HD, only 1–4% of striatal neurons in all grades of HD have nuclear aggregates. Neuropil aggregates, which we have identified by electron microscopy to occur in dendrites and dendritic spines, could play a role in the known dendritic pathology that occurs in HD. Aggregates increase in size in advanced grades, suggesting that they may persist in neurons that are more likely to survive. Ubiquitination is apparent in only a subset of aggregates, suggesting that ubiquitin-mediated proteolysis of aggregates may be late or variable.
CNS Wound Healing Is Severely Depressed in Metallothionein I- and II-Deficient Mice
Authors: Penkowa Milena , Carrasco Javier , Giralt Mercedes , Moos Torben , Hidalgo Juan
Published: April 2018

To characterize the physiological role of metallothioneins I and II (MT-I+II) in the brain, we have examined the chronological effects of a freeze injury to the cortex in normal and MT-I+II null mice. In normal mice, microglia/macrophage activation and astrocytosis were observed in the areas surrounding the lesion site, peaking at ∼1 and 3 d postlesion (dpl), respectively. At 20 dpl, the parenchyma had regenerated. Both brain macrophages and astrocytes surrounding the lesion increased the MT-I+II immunoreactivity, peaking at ∼3 dpl, and at 20 dpl it was similar to that of unlesioned mice.In situhybridization analysis indicates that MT-I+II immunoreactivity reflects changes in the messenger levels. In MT-I+II null mice, microglia/macrophages infiltrated the lesion heavily, and at 20 dpl they were still present. Reactive astrocytosis was delayed and persisted at 20 dpl. In contrast to normal mice, at 20 dpl no wound healing had occurred. The rate of apoptosis, as determined by using terminal deoxynucleotidyl transferase-mediated dUTP–biotin nick end labeling, was drastically increased in neurons of ipsilateral cortex of the MT-I+II null mice. Our results demonstrate that MT-I+II are essential for a normal wound repair in the CNS, and that their deficiency impairs neuronal survival.
Noninvasive Measurements of the Membrane Potential and GABAergic Action in Hippocampal Interneurons
Authors: Verheugen Jos A. H. , Fricker Desdemona , Miles Richard
Published: April 2018

Neurotransmitters affect the membrane potential (Vm) of target cells by modulating the activity of receptor-linked ion channels. The direction and amplitude of the resulting transmembrane current depend on the resting level ofVmand the gradient across the membrane of permeant ion species.Vm, in addition, governs the activation state of voltage-gated channels. Knowledge of the exact level ofVmis therefore crucial to evaluate the nature of the neurotransmitter effect. However, the traditional methods to measureVm, with microelectrodes or the whole-cell current-clamp technique, have the drawback that the recording pipette is in contact with the cytoplasm, and dialysis with the pipette solution alters the ionic composition of the interior of the cell. Here we describe a novel technique to determine theVmof an intact cell from the reversal potential of K+currents through a cell-attached patch. Applying the method to interneurons in hippocampal brain slices yielded more negative values forVmthan subsequent whole-cell current-clamp measurements from the same cell, presumably reflecting the development of a Donnan potential between cytoplasm and pipette solution in the whole-cell mode. Cell-attachedVmmeasurements were used to study GABAergic actions in intact CA1 interneurons. In 1- to 3-week-old rats, bath-applied GABA inhibited these cells by stabilizingVmat a level depending on contributions from both GABAAand GABABcomponents. In contrast, in 1- to 4-d-old animals, only GABAAreceptors were activated resulting in a depolarizing GABA response.
Sonic Hedgehog Promotes Neuronal Differentiation of Murine Spinal Cord Precursors and Collaborates with Neurotrophin 3 to Induce Islet-1
Authors: Dutton Renée , Yamada Toshiya , Turnley Ann , Bartlett Perry F. , Murphy Mark
Published: April 2018

Sonic hedgehog (Shh) is strongly implicated in the development of ventral structures in the nervous system. Addition of Sonic hedgehog protein to chick spinal cord explants induces floor plate and motoneuron development. Whether Shh acts directly to induce these cell types or whether their induction is mediated by additional factors is unknown. To further investigate the role of Shh in spinal neuron development, we have used low-density cultures of murine spinal cord precursor cells. Shh stimulated neuronal differentiation; however, it did not increase the proportion of neurons expressing the first postmitotic motoneuron marker Islet-1. Moreover, Shh did induce Islet-1 expression in neural tube explants, suggesting that it acts in combination with neural tube factors to induce motoneurons. Another factor implicated in motoneuron development is neurotrophin 3 (NT3), and when assayed in isolated precursor cultures, it had no effect on Islet-1 expression. However, the combination of N-terminal Shh and NT3 induced Islet-1 expression in the majority of neurons in low-density cultures of caudal intermediate neural plate. Furthermore, in explant cultures, Shh-mediated Islet-1 expression was blocked by an anti-NT3 antibody. Previous studies have shown expression of NT3 in the region of motoneuron differentiation and that spinal fusimotor neurons are lost in NT3 knock-out animals. Taken together, these findings suggest that Shh can act directly on spinal cord precursors to promote neuronal differentiation, but induction of Islet-1 expression is regulated by factors additional to Shh, including NT3.
Topographic Organization of Human Visual Areas in the Absence of Input from Primary Cortex
Authors: Baseler Heidi A. , Morland Antony B. , Wandell Brian A.
Published: April 2018

Recently, there has been evidence for considerable plasticity in primary sensory areas of adult cortex. In this study, we asked to what extent topographical maps in human extrastriate areas reorganize after damage to a portion of primary visual (striate) cortex, V1. Functional magnetic resonance imaging signals were measured in a subject (G.Y.) with a large calcarine lesion that includes most of primary visual cortex but spares the foveal representation. When foveal stimulation was present, intact cortex in the lesioned occipital lobe exhibited conventional retinotopic organization. Several visual areas could be identified (V1, V2, V3, V3 accessory, and V4 ventral). However, when stimuli were restricted to the blind portion of the visual field, responses were found primarily in dorsal extrastriate areas. Furthermore, cortex that had formerly shown normal topography now represented only the visual field around the lower vertical meridian. Several possible sources for this reorganized activity are considered, including transcallosal connections, direct subcortical projections to extrastriate cortex, and residual inputs from V1 near the margin of the lesion. A scheme is described to explain how the reorganized signals could occur based on changes in the local neural connections.
Cortical Visuomotor Integration during Eye Pursuit and Eye–Finger Pursuit
Authors: Nishitani Nobuyuki , Uutela Kimmo , Shibasaki Hiroshi , Hari Riitta
Published: April 2018

To elucidate cortical mechanisms of visuomotor integration, we recorded whole-scalp neuromagnetic signals from six normal volunteers while they were viewing a black dot moving linearly at the speed of 4°/sec within a virtual rectangle. The dot changed its direction randomly once every 0.3–2 sec. The subject either (1) fixated a cross in the center of the screen (eye fixation task), (2) followed the moving dot with the eyes (eye pursuit task), or (3) followed the dot with both the eyes and the right index finger (eye–finger pursuit task). Prominent magnetic signals, triggered by the changes of the direction of the dot, were seen in all conditions, but they were clearly enhanced by the tasks and were strongest during the eye–finger pursuit task and over the anterior inferior parietal lobule (aIPL). Source modeling indicated activation of aIPL [Brodmann’s area (BA) 40], the posterosuperior parietal lobule (SPL; BA 7), the dorsolateral frontal cortex (DLF; BA 6), and the occipital cortex (BA 18/19). The activation first peaked in the occipital areas, then in the aIPL and DLF, and some 50 msec later in the SPL. Our results suggest that all these areas are involved in visuomotor transformation, with aIPL playing a crucial role in this process.
Functionally Independent Components of the Late Positive Event-Related Potential during Visual Spatial Attention
Authors: Makeig Scott , Westerfield Marissa , Jung Tzyy-Ping , Covington James , Townsend Jeanne , Sejnowski Terrence J. , Courchesne Eric
Published: April 2018

Human event-related potentials (ERPs) were recorded from 10 subjects presented with visual target and nontarget stimuli at five screen locations and responding to targets presented at one of the locations. The late positive response complexes of 25–75 ERP average waveforms from the two task conditions were simultaneously analyzed with Independent Component Analysis, a new computational method for blindly separating linearly mixed signals. Three spatially fixed, temporally independent, behaviorally relevant, and physiologically plausible components were identified without reference to peaks in single-channel waveforms. A novel frontoparietal component (P3f) began at ∼140 msec and peaked, in faster responders, at the onset of the motor command. The scalp distribution of P3f appeared consistent with brain regions activated during spatial orienting in functional imaging experiments. A longer-latency large component (P3b), positive over parietal cortex, was followed by a postmotor potential (Pmp) component that peaked 200 msec after the button press and reversed polarity near the central sulcus. A fourth component associated with a left frontocentral nontarget positivity (Pnt) was evoked primarily by target-like distractors presented in the attended location. When no distractors were presented, responses of five faster-responding subjects contained largest P3f and smallest Pmp components; when distractors were included, a Pmp component appeared only in responses of the five slower-responding subjects. Direct relationships between component amplitudes, latencies, and behavioral responses, plus similarities between component scalp distributions and regional activations reported in functional brain imaging experiments suggest that P3f, Pmp, and Pnt measure the time course and strength of functionally distinct brain processes.
A Neurocomputational Theory of the Dopaminergic Modulation of Working Memory Functions
Authors: Durstewitz Daniel , Kelc Marian , Güntürkün Onur
Published: April 2018

The dopaminergic modulation of neural activity in the prefrontal cortex (PFC) is essential for working memory. Delay-activity in the PFC in working memory tasks persists even if interfering stimuli intervene between the presentation of the sample and the target stimulus. Here, the hypothesis is put forward that the functional role of dopamine in working memory processing is to stabilize active neural representations in the PFC network and thereby to protect goal-related delay-activity against interfering stimuli. To test this hypothesis, we examined the reported dopamine-induced changes in several biophysical properties of PFC neurons to determine whether they could fulfill this function. An attractor network model consisting of model neurons was devised in which the empirically observed effects of dopamine on synaptic and voltage-gated membrane conductances could be represented in a biophysically realistic manner. In the model, the dopamine-induced enhancement of the persistent Na+and reduction of the slowly inactivating K+current increased firing of the delay-active neurons, thereby increasing inhibitory feedback and thus reducing activity of the “background” neurons. Furthermore, the dopamine-induced reduction of EPSP sizes and a dendritic Ca2+current diminished the impact of intervening stimuli on current network activity. In this manner, dopaminergic effects indeed acted to stabilize current delay-activity. Working memory deficits observed after supranormal D1-receptor stimulation could also be explained within this framework. Thus, the model offers a mechanistic explanation for the behavioral deficits observed after blockade or after supranormal stimulation of dopamine receptors in the PFC and, in addition, makes some specific empirical predictions.
Central Neuronal Circuit Innervating the Lordosis-Producing Muscles Defined by Transneuronal Transport of Pseudorabies Virus
Authors: Daniels Derek , Miselis Richard R. , Flanagan-Cato Loretta M.
Published: April 2018

The lordosis reflex is a hormone-dependent behavior displayed by female rats during mating. This study used the transneuronal tracer pseudorabies virus (PRV) to investigate the CNS network that controls the lumbar epaxial muscles that produce this posture. After PRV was injected into lumbar epaxial muscles, the time course analysis of CNS viral infection showed progressively more PRV-labeled neurons in higher brain structures after longer survival times. In particular, the medullary reticular formation, periaqueductal gray (PAG), and ventromedial nucleus of the hypothalamus (VMN) were sequentially labeled with PRV, which supports the proposed hierarchical network of lordosis control. Closer inspection of the PRV-immunoreactive neurons in the PAG revealed a marked preponderance of spheroid neurons, rather than fusiform or triangular morphologies. Furthermore, PRV-immunoreactive neurons were concentrated in the ventrolateral column, rather than the dorsal, dorsolateral, or lateral columns of the PAG. Localization of the PRV-labeled neurons in the VMN indicated that the majority were located in the ventrolateral subdivision, although some were also in other subdivisions of the VMN. As expected, labeled cells also were found in areas traditionally associated with sympathetic outflow to blood vessels and motor pathways, including the intermediolateral nucleus of the spinal cord, the paraventricular hypothalamic nucleus, the red nucleus, and the motor cortex. These results suggest that the various brain regions along the neuraxis previously implicated in the lordosis reflex are indeed serially connected.
Lateralized Effects of Medial Prefrontal Cortex Lesions on Neuroendocrine and Autonomic Stress Responses in Rats
Authors: Sullivan Ron M. , Gratton Alain
Published: April 2018

The medial prefrontal cortex (mPFC) is highly activated by stress and modulates neuroendocrine and autonomic function. Dopaminergic inputs to mPFC facilitate coping ability and demonstrate considerable hemispheric functional lateralization. The present study investigated the potentially lateralized regulation of stress responses at the level of mPFC output neurons, using ibotenic acid lesions. Neuroendocrine function was assessed by plasma corticosterone increases in response to acute or repeated 20 min restraint stress. The primary index of autonomic activation was gastric ulcer development during a separate cold restraint stress. Restraint-induced defecation was also monitored. Plasma corticosterone levels were markedly lower in response to repeated versus acute restraint stress. In acutely restrained animals, right or bilateral, but not left mPFC lesions, decreased prestress corticosterone levels, whereas in repeatedly restrained rats, the same lesions significantly reduced the peak stress-induced corticosterone response. Stress ulcer development (after a single cold restraint stress) was greatly reduced by either right or bilateral mPFC lesions but was unaffected by left lesions. Restraint-induced defecation was elevated in animals with left mPFC lesions. Finally, a left-biased asymmetry in adrenal gland weights was observed across animals, which was unaffected by mPFC lesions. The results suggest that mPFC output neurons demonstrate an intrinsic right brain specialization in both neuroendocrine and autonomic activation. Such findings may be particularly relevant to clinical depression which is associated with both disturbances in stress regulatory systems and hemispheric imbalances in prefrontal function.
Properties and Plasticity of Excitatory Synapses on Dopaminergic and GABAergic Cells in the Ventral Tegmental Area
Authors: Bonci Antonello , Malenka Robert C.
Published: April 2018

Excitatory inputs to the ventral tegmental area (VTA) influence the activity of both dopaminergic (DA) and GABAergic (GABA) cells, yet little is known about the basic properties of excitatory synapses on these two cell types. Using a midbrain slice preparation and whole-cell recording techniques, we found that excitatory synapses on DA and GABA cells display several differences. Synapses on DA cells exhibit a depression in response to repetitive activation, are minimally affected by the GABABreceptor agonist baclofen, and express NMDA receptor-dependent long-term potentiation (LTP). In contrast, synapses on GABA cells exhibit a facilitation in response to repetitive activation, are depressed significantly by baclofen, and do not express LTP. The relative contribution of NMDA and non-NMDA receptors to the synaptic currents recorded from the two cell types is the same as is the depression of synaptic transmission elicited by the application of adenosine, serotonin, or methionine enkephalin (met-enkephalin). The significant differences in the manner in which excitatory synaptic inputs to DA and GABA cells in the VTA can be modulated have potentially important implications for understanding the behavior of VTA neurons during normal behavior and during pathological states such as addiction.
CD95 Ligand (Fas-L/APO-1L) and Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Mediate Ischemia-Induced Apoptosis in Neurons
Authors: Martin-Villalba Ana , Herr Ingrid , Jeremias Irmela , Hahne Michael , Brandt Roland , Vogel Johannes , Schenkel Johannes , Herdegen Thomas , Debatin Klaus-Michael
Published: April 2018

Programmed cell death plays an important role in the neuronal degeneration after cerebral ischemia, but the underlying mechanisms are not fully understood. Here we examined,in vivoandin vitro, whether ischemia-induced neuronal death involves death-inducing ligand/receptor systems such as CD95 and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). After reversible middle cerebral artery occlusion in adult rats, both CD95 ligand and TRAIL were expressed in the apoptotic areas of the postischemic brain. Further recombinant CD95 ligand and TRAIL proteins induced apoptosis in primary neurons and neuron-like cellsin vitro. The immunosuppressant FK506, which most effectively protects against ischemic neurodegeneration, prevented postischemic expression of these death-inducing ligands bothin vivoandin vitro. FK506 also abolished phosphorylation, but not expression, of the c-Jun transcription factor involved in the transcriptional control of CD95 ligand. Most importantly, inlprmice expressing dysfunctional CD95, reversible middle cerebral artery occlusion resulted in infarct volumes significantly smaller than those found in wild-type animals. These results suggest an involvement of CD95 ligand and TRAIL in the pathophysiology of postischemic neurodegeneration and offer alternative strategies for the treatment of cardiovascular brain disease.
Nitric Oxide Stimulates cGMP Production and Mimics Synaptic Responses in Metacerebral Neurons ofAplysia
Authors: Koh Hae-Young , Jacklet Jon W.
Published: April 2018

Nitric oxide (NO) acts as a neurotransmitter and neuromodulator in the nervous systems of many vertebrates and invertebrates. We investigated the mechanism of NO action at an identified synapse between a mechanoafferent neuron, C2, and the serotonergic metacerebral cell (MCC) in the cerebral ganglion of the molluscAplysia californica. Stimulation of C2 produces a decreasing conductance, very slow EPSP in the MCC. C2 is thought to use histamine and NO as cotransmitters at this synapse, because both agents mimic the membrane responses. Now we provide evidence that treatment with NO donors stimulates soluble guanylyl cyclase (sGC) in the MCC, and as a result cGMP increases.S-Nitrosocysteine (SNC, an NO donor) and 8-bromo-cGMP (8-Br-cGMP) both induced the membrane depolarization and increase in input resistance that are characteristic of the very slow EPSP. Two inhibitors of sGC, 6-anilino-5,8-quinolinequinone (LY83583) and 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), suppressed both the very slow EPSP and the membrane responses to SNC but not the histamine membrane responses. NO-induced cGMP production was determined in the MCC using cGMP immunocytochemistry (cGMP-IR). In the presence of 3-isobutyl-1-methylxanthine (IBMX), 10 μmSNC was sufficient to induce cGMP-IR, and the staining intensity increased as the SNC dose was increased. This cGMP-IR was suppressed by ODQ in a dose-dependent manner and completely blocked by 10 μmODQ. Histamine did not induce cGMP-IR. The results suggest that NO stimulates sGC-dependent cGMP synthesis in the MCC and that cGMP mediates the membrane responses. The cotransmitter histamine induces essentially the same membrane responses but seems to use a separate and distinct second messenger pathway.