bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabotropic glutamate receptor mRNA expression in the basal ganglia of the rat
Authors: Testa CM , Standaert DG , Young AB , Penney JB
Published: April 2018

Metabotropic glutamate receptors (mGluRs) couple the actions of glutamate to intracellular second messenger systems through G-proteins. The mGluRs play an important role in the regulation of basal ganglia function. Ligand binding studies have revealed that the basal ganglia contain at least two pharmacological types of metabotropic binding sites. Agonists of mGluRs can affect both in vitro electrophysiologic responses of striatal neurons and motor behavior in vivo. Recently, cDNAs encoding five mGluRs have been cloned, each with distinct structural and pharmacological properties. In order to elucidate the function of these receptors in the biology of the extrapyramidal motor system, we have used in situ hybridization to examine the regional and cellular expression patterns of mGluR1-mGluR5 in the adult rat basal ganglia. In the striatum, all of these mGluRs were present in widely varying relative densities and cellular patterns. MGluR5 was particularly prominent, and exhibited a heterogeneous cellular distribution, with labeled and unlabeled populations of neurons. MGluR2 was expressed in a small population of large polygonal striatal neurons. The subthalamic nucleus was the only other basal ganglia structure that expressed mGluR2. Distinct cellular distributions of mGluR expression were also observed within the nucleus accumbens, globus pallidus, ventral pallidum, and substantia nigra pars reticulata. MGluR3 was expressed in glia in all basal ganglia structures, but was observed in neurons only in the striatum, substantia nigra pars reticulata, and very weakly in the subthalamic nucleus. Comparison of the restricted mGluR2 and mGluR3 mRNA distributions with that of metabotropic ligand binding sites supports a possible presynaptic location for these receptors in the basal ganglia. MGluR1 was the only mGluR message prominently expressed in the dopaminergic neurons of the substantia nigra pars compacta, suggesting the involvement of this receptor in the regulation of dopamine release from nigrostriatal terminals.
Target tissues and innervation regulate the characteristics of K+ currents in chick ciliary ganglion neurons developing in situ
Authors: Dourado MM , Brumwell C , Wisgirda ME , Jacob MH , Dryer SE
Published: April 2018

The expression of appropriate ensembles of ionic channels is necessary for the differentiation and normal function of vertebrate neurons. Cell- cell interactions may regulate the expression and properties of ionic channels in embryonic neurons. Previous studies have shown that the expression of A-type K+ channels (IA) and Ca2+-activated K+ channels (lK[Ca]) is abnormal in chick ciliary ganglion neurons developing in vitro in the absence of normal cell-cell interactions. Other voltage- activated currents develop normally under these conditions. The present studies were designed to establish the role of the target tissues and the preganglionic innervation in regulating the expression of these currents in embryonic chick ciliary ganglion neurons developing in situ. Surgical manipulations were used to remove the developing optic vesicle, which contains the target tissues, the mid-dorsal region of the midbrain primordium, which contains the preganglionic nucleus, or both, all prior to the formation of the ciliary ganglion. IA and IK[Ca] were then examined in acutely isolated neurons that developed in ovo in the presence (OV+) or absence (OV-) of the normal target tissues, in the presence (MB+) or absence (MB-) of preganglionic innervation, and in the absence of both preganglionic innervation and target tissues (OV- /MB-). The amplitude of IA was unaffected by the operations. However, the activation and inactivation kinetics of IA were two- to threefold faster in OV- or OV-/MB- cells compared to neurons isolated from control OV+ ganglia at embryonic days 11–14 (E11-E14). There were no changes in the voltage dependence of activation or steady-state inactivation, or in the time course of recovery from inactivation. By contrast, neurons isolated from MB- ganglia expressed an IA with amplitude, voltage dependence, and kinetics that were indistinguishable from those of control MB+ and OV+ ganglia. Therefore, interactions with target tissues in the eye play a role in determining the characteristics of IA in developing ciliary ganglion neurons, whereas preganglionic innervation does not. Furthermore, the amplitude of IK[Ca] was reduced by 90–100% in OV-, MB-, and OV-/MB- neurons isolated at E12-E14 as compared to MB+ and OV+ controls. Voltage-activated Ca2+ currents were present at normal amplitudes in all of these neurons. Thus, the expression of IK[Ca] in chick ciliary ganglion neurons is regulated by both target tissue interactions and preganglionic innervation. Therefore, cell-cell interactions are necessary for the expression of a normal ensemble of ionic channels in chick ciliary ganglion neurons developing in situ.
Regional and developmental heterogeneity in splicing of the rat brain NMDAR1 mRNA
Authors: Laurie DJ , Seeburg PH
Published: April 2018

Developmental and regional alternative splicing of the NMDAR1 subunit gene transcript was examined by in situ hybridization in the developing and adult rat brain. NMDAR1 mRNA, barely detectable at embryonic day 14, increased gradually during development until the third postnatal week, after which it declined slightly to adult levels, when it was detected in every examined neuronal type. Each splice form of the primary NMDAR1 gene transcript was found to follow a parallel profile of abundance in the brain, but marked regional differences were observed in splicing at both 5′ and 3′ sequences. The individual regional distributions of splice forms appeared to be established around birth, with little change thereafter, except in the overall abundance. The NMDAR1-a and NMDAR1–2 splice forms occurred extensively and approximately homogeneously throughout brain gray matter. The NMDAR1-b variant was found primarily in the sensorimotor cortex, neonatal lateral caudate, thalamus, hippocampal CA3 field, and cerebellar granule cells, but was absent from adult caudate. The NMDAR1–1 and -4 splice forms were detected in almost complementary patterns; the former was concentrated in more rostral structures such as cortex, caudate, and hippocampus, while the latter was principally in more caudal regions such as thalamus, colliculi, and cerebellum. These two splice forms accounted for a greater proportion of the adult NMDAR1 mRNA than that of the neonate. The NMDAR1–3 mRNA variant was scarce, being detected only at very low levels in postnatal cortex and hippocampus. The different splice forms may generate regional differences in NMDA receptor properties during development and in the adult CNS.
Adaptive representation of dynamics during learning of a motor task
Authors: Shadmehr R , Mussa-Ivaldi FA
Published: April 2018

We investigated how the CNS learns to control movements in different dynamical conditions, and how this learned behavior is represented. In particular, we considered the task of making reaching movements in the presence of externally imposed forces from a mechanical environment. This environment was a force field produced by a robot manipulandum, and the subjects made reaching movements while holding the end-effector of this manipulandum. Since the force field significantly changed the dynamics of the task, subjects' initial movements in the force field were grossly distorted compared to their movements in free space. However, with practice, hand trajectories in the force field converged to a path very similar to that observed in free space. This indicated that for reaching movements, there was a kinematic plan independent of dynamical conditions. The recovery of performance within the changed mechanical environment is motor adaptation. In order to investigate the mechanism underlying this adaptation, we considered the response to the sudden removal of the field after a training phase. The resulting trajectories, named aftereffects, were approximately mirror images of those that were observed when the subjects were initially exposed to the field. This suggested that the motor controller was gradually composing a model of the force field, a model that the nervous system used to predict and compensate for the forces imposed by the environment. In order to explore the structure of the model, we investigated whether adaptation to a force field, as presented in a small region, led to aftereffects in other regions of the workspace. We found that indeed there were aftereffects in workspace regions where no exposure to the field had taken place; that is, there was transfer beyond the boundary of the training data. This observation rules out the hypothesis that the subject's model of the force field was constructed as a narrow association between visited states and experienced forces; that is, adaptation was not via composition of a look-up table. In contrast, subjects modeled the force field by a combination of computational elements whose output was broadly tuned across the motor state space. These elements formed a model that extrapolated to outside the training region in a coordinate system similar to that of the joints and muscles rather than end-point forces. This geometric property suggests that the elements of the adaptive process represent dynamics of a motor task in terms of the intrinsic coordinate system of the sensors and actuators.
Localization of calcium signals by a mobile calcium buffer in frog saccular hair cells
Authors: Roberts WM
Published: April 2018

A recent study (Roberts, 1993) of saccular hair cells from grass frogs (Rana pipiens) has suggested a mechanism by which the unusually high concentrations of calcium-binding proteins found in certain sensory receptors and neurons, particularly in the auditory system, can influence short-range intracellular calcium signaling. In frog saccular hair cells, the mechanism operates within arrays of calcium channels and calcium-activated potassium channels that are involved in the cells' electrical resonance and synaptic transmission. The present study tests the hypothesis that calbindin-D28k, one of the most abundant proteins in these cells, can serve as a mobile calcium buffer that reduces and localizes changes in the intracellular free-calcium concentration ([Ca2+]i) by shuttling calcium away from the channel arrays. Based upon theoretical analysis and computer modeling, it is shown that [Ca2+]i near one or more open channels quickly reaches a steady-state level determined primarily by two properties of the buffer, the mean time (tau c) before it captures a free-calcium ion and a replenishment factor (R), which are related to the buffer's diffusional mobility (DBu), association rate constant (kon), and concentration (Bo) by tau c = (konB0)-1 and R = B0DBu. Simulation of calcium entry through a channel array showed that approximately 1.5 mM of a molecule with the diffusional and binding properties expected for calbindin-D28k (Bo approximately 8 mM calcium-binding sites) is needed to reproduce the previous experimental results. A lower concentration (B0 = 2 mM) was almost completely depleted within the channel array by a modest calcium current (8 pA = 12% of calcium channels open), but still had two important effects: it caused [Ca2+]i to fall steeply with distance outside the array (space constant < 50 nm), and returned [Ca2+]i quickly to the resting level after the channels closed. A high concentration of calbindin-D28k can thus influence the cell's electrical resonance and synaptic transmission. Its most important functions may be to localize regions of high [Ca2+]i and speed the return of [Ca2+]i toward the resting level.
Analysis of agonist and antagonist activities of phenylglycine derivatives for different cloned metabotropic glutamate receptor subtypes
Authors: Hayashi Y , Sekiyama N , Nakanishi S , Jane DE , Sunter DC , Birse EF , Udvarhelyi PM , Watkins JC
Published: April 2018

The metabotropic glutamate receptors (mGluRs) consist of at least seven different subtypes and are coupled to intracellular signal transduction via G proteins. However, the lack of specific antagonists for the mGluRs limited the precise characterization of the role of the individual mGluRs. In this study, we investigated the agonist and antagonist activities of a series of phenylglycine derivatives for the mGluRs by examining their effects on the signal transduction of representative mGluR1, mGluR2, and mGluR4 subtypes expressed individually in Chinese hamster ovary cells. The phenylglycine derivatives examined included (S)- and (R)-forms of 3- hydroxyphenylglycine (3HPG), 4-carboxy-phenylglycine (4CPG), 4-carboxy- 3-hydroxyphenylglycine (4C3HPG), 3-carboxy-4-hydroxyphenylglycine (3C4HPG), and (+)- and (-)-alpha-methyl-4-carboxyphenylglycine (alpha M4CPG). Among these 10 compounds, (S)-3HPG acted as an agonist for mGluR1, while (S)-4C3HPG, (S)-3C4HPG, and (S)-4CPG served as effective agonists for mGluR2. The rank order of agonist potencies for mGluR2 was L-glutamate > (S)-4C3HPG > (S)-3C4HPG > (S)-4CPG. No other phenylglycine derivatives showed any definite agonist activity on either mGluR1 or mGluR2. Among the phenylglycine derivatives with no mGluR1 agonist activity, (S)-4C3HPG, (S)-3C4HPG, (S)-4CPG, and (+)- alpha M4CPG effectively antagonized the action of L-glutamate on mGluR1. The rank order of antagonist potencies was (S)-4C3HPG > or = (S)-4CPG > or = (+)-alpha M4CPG > (S)-3C4HPG. The Schild plot analysis indicated that (RS)-4C3HPG, (S)-4CPG, and (+)-alpha M4CPG all act as competitive antagonists for mGluR1 with pA2 values of 4.38, 4.46, and 4.38, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Changes in the Electrical Properties of Supraoptic Nucleus Oxytocin and Vasopressin Neurons during Lactation
Authors: Stern Javier E. , Armstrong William E.
Published: April 2018

Magnocellular oxytocin (OT) and vasopressin (VP) neurons adopt different firing patterns in response to relevant physiological stimuli. OT neurons selectively display short (2–4 sec), high-frequency bursts of action potentials that are highly synchronized and correlated with OT release during lactation. The present experiments were done to determine whether the electrophysiological properties of OT neurons differ from those of VP neurons, and whether these properties are modulated during lactation to support short bursting activity. Intracellular recordingsin vitrowere obtained from immunochemically identified supraoptic neurons of diestrous or lactating female rats. Resting membrane potential, input resistance, membrane time constant, and the depolarizing afterpotential did not differ among groups. However, near spike threshold, OT, but not VP, neurons expressed a sustained outward rectification that was removed by small hyperpolarizing pulses and a rebound depolarization that occurred at the offset of these hyperpolarizing pulses. The rebound depolarization was short (<2 sec), supported brief bursts of action potentials, and was significantly larger during lactation. Neurons expressing the outward rectification also exhibited strong spike frequency adaptation during prolonged (1–4 sec) depolarization. Spike width, the Ca2+-dependent afterhyperpolarization, and the degree of spike broadening of OT, but not VP, neurons were also larger during lactation, suggesting an increase in Ca2+influx per spike. The results indicate that OT neurons possess properties favoring the expression of short spike trains, and that some of these properties are enhanced during lactation. In addition, spikes in OT neurons may promote more Ca2+influx in this state.
Vicious Cycle Involving Na+Channels, Glutamate Release, and NMDA Receptors Mediates Delayed Neurodegeneration through Nitric Oxide Formation
Authors: Strijbos Paul J. L. M. , Leach Michael J. , Garthwaite John
Published: April 2018

The mechanisms by which neurons die after cerebral ischemia and related conditionsin vivoare unclear, but they are thought to involve voltage-dependent Na+channels, glutamate receptors, and nitric oxide (NO) formation because selective inhibition of each provides neuroprotection. It is not known precisely what their roles are, nor whether they interact within a single cascade or in parallel pathways. These questions were investigated using anin vitroprimary cell culture model in which striatal neurons undergo a gradual and delayed neurodegeneration after a brief (5 min) challenge with the glutamate receptor agonist NMDA. Unexpectedly, NO was generated continuously by the cultures for up to 16 hr after the NMDA exposure. Neuronal death followed the same general time course except that its start was delayed by ∼4 hr. Application of the NO synthase inhibitor nitroarginine after, but not during, the NMDA exposure inhibited NO formation and protected against delayed neuronal death. Blockade of NMDA receptors or of voltage-sensitive Na+channels [with tetrodotoxin (TTX)] during the postexposure period also inhibited both NO formation and cell death. The NMDA exposure resulted in a selective accumulation of glutamate in the culture medium during the period preceding cell death. This glutamate release could be inhibited by NMDA antagonism or by TTX, but not by nitroarginine. These data suggest that Na+channels, glutamate receptors, and NO operate interdependently and sequentially to cause neurodegeneration. At the core of the mechanism is a vicious cycle in which NMDA receptor stimulation causes activation of TTX-sensitive Na+channels, leading to glutamate release and further NMDA receptor stimulation. The output of the cycle is an enduring production of NO from neuronal sources, and this is responsible for delayed neuronal death. The same neurons, however, could be induced to undergo more rapid NMDA receptor-dependent death that required neither TTX-sensitive Na+channels nor NO.
Kainate Binding Proteins Possess Functional Ion Channel Domains
Authors: Villmann Carmen , Bull Leonard , Hollmann Michael
Published: April 2018

Kainate binding proteins (KBPs) are highly homologous to ionotropic glutamate receptors; however, no ion channel function has been demonstrated for these proteins. To investigate possible reasons for the apparent lack of ion channel function we transplanted the ion channel domains of five KBPs into glutamate receptors GluR 6 and GluR1. In each case we obtained functional chimeric receptors in which glutamatergic agonists were able to open the KBP-derived ion channel with EC50values identical to those of the subunit contributing the ligand binding domain. Maximal current amplitudes were significantly smaller than those of the parent clones, however. We also show that the KBP ion channels are highly permeable for calcium and have certain pharmacological properties that are distinct from all other glutamate receptor (GluR) subunits. Thus, all five known KBPs, in addition to their well characterized functional ligand binding sites, have functional ion permeation pathways. Our data suggest that the lack of ion channel function in wild-type KBPs results from a failure to translate ligand binding into channel opening. We interpret our findings to indicate the requirement for a modulatory protein or an additional subunit serving to alter the structure of the KBP subunit complex such that signal transduction is enabled from the ligand binding site to the intrinsically functional ion pore.
Increased Susceptibility to Ischemic Brain Damage in Transgenic Mice Overexpressing the Amyloid Precursor Protein
Authors: Zhang Fangyi , Eckman Chris , Younkin Steven , Hsiao Karen K. , Iadecola Costantino
Published: April 2018

We studied the role of the amyloid precursor protein (APP) in ischemic brain damage using transgenic mice overexpressing APP. The middle cerebral artery (MCA) was occluded in FVB/N mice expressing APP695.SWE (Swedish mutation) and in nontransgenic littermates. Infarct volume (cubic millimeters) was assessed 24 hr later in thionin-stained brain sections. The infarct produced by MCA occlusion was enlarged in the transgenics (+32 ± 6%;n= 12;p< 0.05;ttest). Measurement of APP by ELISA revealed that, although relatively high levels of Aβ were present in the brain of the transgenics (Aβ1–40= 80 ± 19 pmol/g;n= 6), there were no differences between ischemic and nonischemic hemispheres (p> 0.05). The reduction in cerebral blood flow produced by MCA occlusion at the periphery of the ischemic territory was more pronounced in APP transgenics (−42 ± 8%;n= 9) than in controls (−20 ± 8%;n= 9). Furthermore, the vasodilatation produced by neocortical application of the endothelium-dependent vasodilator acetylcholine (10 μm) was reduced by 82 ± 5% (n= 8;p< 0.05) in APP transgenics. The data demonstrate that APP overexpression increases the susceptibility of the brain to ischemic injury. The effect is likely to involve the Aβ-induced disturbance in endothelium-dependent vascular reactivity that leads to more severe ischemia in regions at risk for infarction. The cerebral vascular actions of peptides deriving from APP metabolism may play a role in the pathogenic effects of APP.
Adenine Nucleotides Undergo Rapid, Quantitative Conversion to Adenosine in the Extracellular Space in Rat Hippocampus
Authors: Dunwiddie Thomas V. , Diao Lihong , Proctor William R.
Published: April 2018

There are multiple mechanisms by which adenine nucleotides can be released into the extracellular space in brain. Adenine nucleotides are converted extracellularly to adenosine, which then acts on adenosine receptors to elicit physiological responses, but the rate at which this conversion takes place is unknown. In the present experiments, adenine nucleotides were applied to individual hippocampal neurons, and the subsequent activation of a postsynaptic K+conductance by adenosine A1receptors was used to determine the rate of adenosine formation. None of the adenine nucleotides tested (cAMP, AMP, ADP, and ATP) activated A1receptors directly at the concentrations tested (≤200 μm). AMP, ADP, and ATP were all rapidly converted to adenosine, with aT1/2for ATP conversion to adenosine of ∼200 msec, and the last step in this pathway (transformation of AMP to adenosine by 5′-nucleotidase) seems to be the rate-limiting step. As we have reported previously, cAMP is converted to adenosine as well, but on a much slower time scale than any of the other nucleotides tested. These experiments demonstrate that fast, localized release of AMP, ADP, or ATP can result in a transient activation of adenosine receptors but that this is unlikely to occur with cAMP. The existence of a highly active ecto-nucleotidase pathway in brain provides a mechanism for the rapid generation of adenosine after the release of adenine nucleotides into the extracellular space.
Expression and Clustered Distribution of an Inwardly Rectifying Potassium Channel, KAB-2/Kir4.1, on Mammalian Retinal Müller Cell Membrane: Their Regulation by Insulin and Laminin Signals
Authors: Ishii Masaru , Horio Yoshiyuki , Tada Yoshihiko , Hibino Hiroshi , Inanobe Atsushi , Ito Minoru , Yamada Mitsuhiko , Gotow Takahiro , Uchiyama Yasuo , Kurachi Yoshihisa
Published: April 2018

Inwardly rectifying potassium (K+) channels (Kir) in Müller cells, the dominant glial cells in the retina, are supposed to be responsible for the spatial buffering action of K+ions. The molecular properties and subcellular localization of Müller cell Kir channels in rat and rabbit retinas were examined by using electrophysiological, molecular biological, and immunostaining techniques. Only a single population of Kir channel activity, the properties of which were identical to those of KAB-2/Kir4.1 expressed in HEK293T cells, could be recorded from endfoot to the distal portion of Müller cells. Consistently, Northern blot,in situhybridization, and RT-PCR analyses indicated expression of Kir4.1 in Müller cells per se. The Kir4.1 immunoreactivity was distributed in clusters throughout Müller cell membrane. The Kir4.1 expression in Müller cells disappeared promptly after culturing. When the dissociated Müller cells were cultured on laminin-coated dishes in the presence of insulin, Kir4.1 immunoreactivity was detected in a clustered manner on the cell membrane. Because insulin and laminin exist in the surrounding of Müller cells in the retina, these substances possibly may be physiological regulators of expression and distribution of Kir4.1 in Müller cellsin vivo.
Correspondence Noise and Signal Pooling in the Detection of Coherent Visual Motion
Authors: Barlow Horace , Tripathy Srimant P.
Published: April 2018

In the random dot kinematograms used to analyze the detection of coherent motion in the middle temporal visual area (MT) and in psychophysical experiments the exact way that dots are paired between successive presentations is not known by the observer. We show how to calculate the limit to coherence threshold caused by this uncertainty, which we call “correspondence noise.” We compare ideal thresholds limited only by this noise with those of human observers when dot density, ratio of dot numbers in two fields, area of stimulus, number of fields, and method of generation of the coherent dots are varied. The observed thresholds vary in the same way as the ideal thresholds over wide ranges, but they are much higher. We think this difference is because the ideal detector takes advantage of the high precision with which dots are placed in the kinematograms, whereas the neural motion system can only operate with low precision. When kinematograms are generated with decreased precision of dot placement, the ideal detector no longer has this advantage, and the gap between ideal and actual performance is greatly reduced. Because the signals that result from objects moving in the real world are scattered over broad ranges of direction and velocity, high precision is not needed, and it is advantageous for the motion system to pool information over broad ranges. Other mismatches between kinematograms and the neural motion system, and internal noise, may also elevate human thresholds relative to the ideal detector. The importance of external noise suggests that the neurons of MT form a vast array of optimal filters, each matched to a different combination of parameters in the multidimensional space required to define motion in patches of the visual field.
Insensitivity of the Hippocampus to Environmental Stimulation during Postnatal Development
Authors: Waters Nicholas S. , Klintsova Anna Y. , Foster Thomas C.
Published: April 2018

Development of cortical sensory systems is influenced by environmental experience during “sensitive periods,” before onset of behavioral function. During these periods, synaptic plasticity is observed, and neuronal function shows increased responsiveness to environmental stimulation. Because the hippocampus is late to develop, and because it demonstrates synaptic plasticity before the onset of behavioral function, this experiment was designed to determine whether, like the sensory cortices, the hippocampus undergoes a period of enhanced responsiveness to the environment. Rats at three ages [postnatal day 16 (P16), P23, and P30] were tested on a hippocampally dependent task, spontaneous alternation, and exposed to a novel environment. They were then killed and processed for immunocytochemistry to Fos or forin vitroelectrophysiology in hippocampal area CA1. Age-matched control subjects were killed immediately after removal from the home cage. Spontaneous alternation was only observed in the oldest (P30) animals. In these same animals, the environmental manipulation resulted in an increase in Fos-like immunoreactivity (FL-IR), relative to controls, and a decrease in the ability to induce long-term potentiation (LTP). In P16 and P23 animals, the environmental manipulation resulted in no differences in hippocampal FL-IR or LTP. These results suggest that, rather than showing increased responsiveness to the environment at these ages, the hippocampus is environmentally insensitive and that it is isolated from the effects of environmental stimuli. The hippocampus, a neural region important for higher cognitive function, may develop via a mechanism different from those observed in the primary sensory cortices.
Dopamine–Adenosine Interactions in the Striatum and the Globus Pallidus: Inhibition of Striatopallidal Neurons through Either D2or A2AReceptors Enhances D1Receptor-Mediated Effects onc-fosExpression
Authors: Le Moine Catherine , Svenningsson Per , Fredholm Bertil B. , Bloch Bertrand
Published: April 2018

D1receptors located on striatonigral neurons and D2receptors located, together with A2Areceptors, on striatopallidal neurons are known to interact functionally. Usingin situhybridization, we examined the effects of D1and D2agonists and of an A2Aantagonist onc-fosmRNA in identified striatal neurons and in globus pallidus. The full D1agonist, SKF 82958 (1 mg/kg), induced a homogenous increase ofc-fosmRNA in the striatum. This increase occurred to a similar extent in D1and D2receptor-containing striatal neurons. Conversely, the D2agonist, quinelorane (2 mg/kg), decreasedc-fosmRNA in these populations but increased it in globus pallidus. The adenosine A2Areceptor antagonist, SCH 58261 (5 mg/kg), also decreasedc-fosmRNA in D2receptor-containing neurons in striatum but did not affect pallidalc-fosmRNA. Concomitant administration of either D1plus D2agonists or D1agonist plus A2Aantagonist caused a potentiation ofc-fosmRNA in striatal neurons expressing the D1receptor and in globus pallidus. However, only the combination of D1and D2agonists modified thec-fosmRNA expression to a “patchy” distribution. Our data show that (1)c-fosexpression can be activated through D1andinhibitedthrough A2Aor D2receptors in both striatal output pathways in normal rats, and (2) D2receptor stimulation as well as A2Areceptor blockade can interact with D1receptor activation to potentiatec-fosexpression in the striatum and the globus pallidus. The data also suggest that the topological alteration ofc-fosexpression after coadministration of D1and D2agonists involves D2receptors located on interneurons or presynaptically on dopaminergic nerve terminals.
Inhibitors of Myosin Light Chain Kinase Block Synaptic Vesicle Pool Mobilization during Action Potential Firing
Authors: Ryan Timothy A.
Published: April 2018

During repetitive action potential firing the maintenance of synaptic transmission relies on a continued supply of synaptic vesicles for fusion with the presynaptic plasma membrane. The mechanism of transport by which vesicles are delivered to the site of fusion from a reserve pool is unknown, as are the biochemical pathways linking intracellular Ca2+elevation with vesicle mobilization. Here, using the fluorescent tracer FM1-43 in hippocampal synaptic terminals, I show that inhibitors of myosin light chain kinase can block mobilization of the reserve pool and not the immediately releasable pool.
The sec6/8 Complex Is Located at Neurite Outgrowth and Axonal Synapse-Assembly Domains
Authors: Hazuka Christopher D. , Foletti Davide L. , Hsu Shu-Chan , Kee Yun , Hopf F. Woodward , Scheller Richard H.
Published: April 2018

The molecules that specify domains on the neuronal plasma membrane for the delivery and accumulation of vesicles during neurite outgrowth and synapse formation are unknown. We investigated the role of the sec6/8 complex, a set of proteins that specifies vesicle targeting sites in yeast and epithelial cells, in neuronal membrane trafficking. This complex was found in layers of developing rat brain undergoing synaptogenesis. In cultured hippocampal neurons, the sec6/8 complex was present in regions of ongoing membrane addition: the tips of growing neurites, filopodia, and growth cones. In young axons, the sec6/8 complex was also confined to periodic domains of the plasma membrane. The distribution of synaptotagmin, synapsin1, sec6, and FM1–43 labeling in cultured neurons suggested that the plasma membrane localization of the sec6/8 complex preceded the arrival of synaptic markers and was downregulated in mature synapses. We propose that the sec6/8 complex specifies sites for targeting vesicles at domains of neurite outgrowth and potential active zones during synaptogenesis.
Interleukin-1 Mediates a Rapid Inflammatory Response After Injection of Adenoviral Vectors into the Brain
Authors: Cartmell Tammy , Southgate Tom , Rees Gwen S. , Castro Maria G. , Lowenstein Pedro R. , Luheshi Giamal N.
Published: April 2018

Adenovirus-mediated gene transfer into the brain is associated with significant inflammation and activation of anti-vector and anti-transgene immune responses that curtail the gene delivery of adenoviruses and therapeutic efficacy. Elucidating the molecular mediators of inflammatory and immune responses to adenoviruses injected into the brain should allow us to inhibit their inflammatory actions, thereby reducing vector clearance and enhance adenoviral-mediated gene transfer into the CNS. Cytokines are primary mediators of the immune response and are released during inflammation. Here we report for the first time that injection of replication-deficient adenovirus vectors into the cerebral ventricles of rats causes a rapid increase in body temperature. This fever response precedes any vector-encoded transgene expression and occurs with vectors encoding no transgene, as well as with vectors encoding a therapeutic transgene i.e., HSV1-thymidine kinase. No fever is detected after infection of the striatum, an important brain target in studies on neurodegeneration. After infection of the brain ventricles, CSF levels of immunoreactive tumor necrosis factor (TNF)-α and interleukin (IL)-1β increase significantly (up to 300-fold). In the hypothalamus, the locus of thermoregulation in the brain, only IL-1β and IL-6 are significantly elevated. A neutralizing TNF-α antibody has no effect on adenovirus-induced fever. However, pretreatment with either the IL-1 receptor antagonist or the cyclooxygenase inhibitor flurbiprofen completely abolishes adenovirus-induced fever, suggesting that IL-1 and prostaglandins are direct mediators of this response. These results are the first to demonstrate that IL-1, but not TNF-α, is the main mediator of a very early inflammatory response to adenovirus in the brain.
Apparent Loss and Hypertrophy of Interneurons in a Mouse Model of Neuronal Ceroid Lipofuscinosis: Evidence for Partial Response to Insulin-Like Growth Factor-1 Treatment
Authors: Cooper Jonathan D. , Messer Anne , Feng Andrew K. , Chua-Couzens Jane , Mobley William C.
Published: April 2018

The neuronal ceroid lipofuscinoses (NCL) are progressive neurodegenerative disorders with onset from infancy to adulthood that are manifested by blindness, seizures, and dementia. In NCL, lysosomes accumulate autofluorescent proteolipid in the brain and other tissues. Themnd/mndmutant mouse was first characterized as exhibiting adult-onset upper and lower motor neuron degeneration, but closer examination revealed early, widespread pathology similar to that seen in NCL. We used the autofluorescent properties of accumulated storage material to map which CNS neuronal populations in themnd/mndmouse show NCL-like pathological changes. Pronounced, early accumulation of autofluorescent lipopigment was found in subpopulations of GABAergic neurons, including interneurons in the cortex and hippocampus. Staining for phenotypic markers normally present in these neurons revealed progressive loss of staining in the cortex and hippocampus ofmnd/mndmice, with pronounced hypertrophy of remaining detectable interneurons. In contrast, even in aged mutant mice, many hippocampal interneurons retained staining for glutamic acid decarboxylase. Treatment with insulin-like growth factor-1 partially restored interneuronal number and reduced hypertrophy in some subregions. These results provide the first evidence for the involvement of interneurons in a mouse model of NCL. Moreover, our findings suggest that at least some populations of these neurons persist in a growth factor-responsive state.
Optical Detection of Synaptically Induced Glutamate Transport in Hippocampal Slices
Authors: Kojima Satoshi , Nakamura Takeshi , Nidaira Takahisa , Nakamura Kyoko , Ooashi Noriko , Ito Etsuro , Watase Kei , Tanaka Kohichi , Wada Keiji , Kudo Yoshihisa , Miyakawa Hiroyoshi
Published: April 2018

Although it has long been believed that glial cells play a major role in transmitter uptake at synapses in the CNS, the relative contribution of glial and neuronal cells to reuptake of synaptically released glutamate has been unclear. Recent identification of the diverse glutamate transporter subtypes provides an opportunity to examine this issue. To monitor glutamate transporter activity, we optically detected synaptically induced changes of membrane potential from hippocampal CA1 field in slice preparations using a voltage-sensitive dye, RH155. In the presence of ionotropic glutamate-receptor blockers, synaptic inputs gave rise to a slow depolarizing response (SDR) in the dendritic field. The amplitude of SDR correlated well with presynaptic activities, suggesting that it was related to transmitter release. The SDR was found to be caused by the activities of glutamate transporters because it was not affected by blockers for GABAA, nACh, 5-HT3, P2X, or metabotropic glutamate receptors but was greatly reduced by dihydrokainate (DHK), a specific blocker for GLT-1 transporter, and byd,l-threo-β-hydroxyaspartate (THA), a blocker for EAAC, GLAST, and GLT-1 transporters. When SDR was detected with RH482 dye, which stains both glial and neuronal cells, 1 mmDHK and 1 mmTHA were equally effective in suppressing SDR. The SDR was very small in GLT-1 knockout mice but was maintained in gerbil hippocampi in which postsynaptic neurons were absent because of ischemia. Because GLT-1 transporters are exclusively expressed in astrocytes, our results provide direct evidence that astrocytes play the dominant role in sequestering synaptically released glutamate.
Role of Primate Superior Colliculus in Preparation and Execution of Anti-Saccades and Pro-Saccades
Authors: Everling Stefan , Dorris Michael C. , Klein Raymond M. , Munoz Douglas P.
Published: April 2018

We investigated how the brain switches between the preparation of a movement where a stimulus is the target of the movement, and a movement where a stimulus serves as a landmark for an instructed movement elsewhere. Monkeys were trained on a pro-/anti-saccade paradigm in which they either had to generate a pro-saccade toward a visual stimulus or an anti-saccade away from the stimulus to its mirror position, depending on the color of an initial fixation point. Neural activity was recorded in the superior colliculus (SC), a structure that is known to be involved in the generation of fast saccades, to determine whether it was also involved in the generation of anti-saccades. On anti-saccade trials, fixation during the instruction period was associated with an increased activity of collicular fixation-related neurons and a decreased activity of saccade-related neurons. Stimulus-related and saccade-related activity was reduced on anti-saccade trials. Our results demonstrate that the anti-saccade task involves (and may require) the attenuation of preparatory and stimulus-related activity in the SC to avoid unwanted pro-saccades. Because the attenuated pre-saccade activity that we found in the SC may be insufficient by itself to elicit correct anti-saccades, additional movement signals from other brain areas are presumably required.
Glutamate Uptake Limits Synaptic Excitation of Retinal Ganglion Cells
Authors: Higgs Matthew H. , Lukasiewicz Peter D.
Published: April 2018

EPSCs of retinal ganglion cells decay more slowly than do those of most other CNS neurons, in part because of the long time course of glutamate release from bipolar cells. Here we investigated how glutamate clearance and AMPA receptor desensitization affect ganglion cell EPSCs in the salamander retinal slice preparation. Inhibition of glutamate uptake greatly prolonged ganglion cell EPSCs evoked by light or monosynaptic electrical stimuli but had little effect on spontaneous miniature EPSCs (mEPSCs). This suggests that single quanta of glutamate are cleared rapidly by diffusion but multiple quanta can interact to lengthen the postsynaptic response. Some interaction between quanta is likely to occur even when glutamate uptake is not inhibited. This seems to depend on quantal content, because reducing glutamate release with low Ca2+, paired-pulse depression, or weak stimuli shortened the EPSC decay. High quantal content glutamate release may lead to desensitization of postsynaptic receptors. We reduced the extent of AMPA receptor desensitization by holding ganglion cells at positive potentials. This increased the amplitude of the late phase of evoked EPSCs but did not affect the decay rate after the first 50 msec of the response. In contrast, the holding potential had little effect on mEPSC kinetics. Our results suggest that desensitization limits the late phase of AMPA receptor–mediated EPSCs, whereas glutamate uptake controls the duration of both AMPA and NMDA receptor–mediated responses.
Nature of the Retrograde Signal from Injured Nerves that Induces Interleukin-6 mRNA in Neurons
Authors: Murphy Patricia G. , Borthwick Lindsay S. , Johnston Robert S. , Kuchel George , Richardson Peter M.
Published: April 2018

In previous studies, interleukin-6 was shown to be synthesized in approximately one-third of lumbar dorsal root ganglion neurons during the first week after nerve transection. In present studies, interleukin-6 mRNA was found to be induced also in axotomized facial motor neurons and sympathetic neurons. The nature of the signal that induces interleukin-6 mRNA in neurons after nerve injury was analyzed. Blocking of retrograde axonal transport by injection of colchicine into an otherwise normal nerve did not induce interleukin-6 mRNA in primary sensory neurons, but injection of colchicine into the nerve stump prevented induction of interleukin-6 mRNA by nerve transection. Therefore, it was concluded that interleukin-6 is induced by an injury factor arising from the nerve stump rather than by interruption of normal retrograde trophic support from target tissues or distal nerve segments. Next, injection into the nerve of a mast cell degranulating agent was shown to stimulate interleukin-6 mRNA in sensory neurons and systemic administration of mast cell stabilizing agents to mitigate the induction of interleukin-6 mRNA in sensory neurons after nerve injury. These data implicate mast cells as one possible source of the factors that lead to induction of interleukin-6 mRNA after nerve injury.

In search of a possible function of inducible interelukin-6, neuronal death after nerve transection was assessed in mice with null deletion of the interleukin-6 gene. Retrograde death of neurons in the fifth lumbar dorsal root ganglion was 45% greater in knockout than in wild-type mice. Thus, endogenous interleukin-6 contributes to the survival of axotomized neurons.
Muscarinic Control of Cytoskeleton in Perisynaptic Glia
Authors: Georgiou John , Robitaille Richard , Charlton Milton P.
Published: April 2018

Similar to astrocytes at CNS synapses, perisynaptic Schwann cells (PSCs) surround nerve terminals at the neuromuscular junction (NMJ). These special teloglial cells are sensitive to neurotransmitters and upregulate glial fibrillary acidic protein (GFAP) when deprived of synaptic activity. We found that activation of muscarinic acetylcholine receptors (mAChRs) at PSCs, but not purinergic (ATP and adenosine) or peptidergic [substance P (SP) and calcitonin gene-related peptide (CGRP)] receptors, prevented this upregulation. When applied onto single PSCs, muscarine evoked Ca2+responses that fatigued but prevented upregulation of this glial cytoskeletal protein. Application of ATP onto single PSCs evoked Ca2+signals that showed little fatigue, and GFAP upregulation occurred. Thus, Ca2+signals alone cannot prevent GFAP upregulation in the PSCs. After blockade of cholinergic receptors by gallamine, neuronal activity was not effective in maintaining low GFAP levels in the perisynaptic glia. Last, immunohistochemistry disclosed mAChRs on PSCs and nearby fibroblasts. Thus, acetylcholine secreted by the nerve terminal acts on the PSCs via mAChRs to regulate GFAP. Cytoskeletal changes may influence perisynaptic glial functions, including growth, remodeling, and modulation of the synapse.
Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB
Authors: Meier Carola , Parmantier Eric , Brennan Angela , Mirsky Rhona , Jessen Kristjan R.
Published: April 2018

Although Schwann cell precursors from early embryonic nerves die in the absence of axonal signals, Schwann cells in older nerves can survive in the absence of axons in the distal stump of transected nerves. This is crucially important, because successful axonal regrowth in a damaged nerve depends on interactions with living Schwann cells in the denervated distal stump. Here we show that Schwann cells acquire the ability to survive without axons by establishing an autocrine survival loop. This mechanism is absent in precursors. We show that insulin-like growth factor, neurotrophin-3, and platelet-derived growth factor-BB are important components of this autocrine survival signal. The secretion of these factors by Schwann cells has significant implications for cellular communication in developing nerves, in view of their known ability to regulate survival and differentiation of other cells including neurons.
Characterization of MALS/Velis-1, -2, and -3: a Family of Mammalian LIN-7 Homologs Enriched at Brain Synapses in Association with the Postsynaptic Density-95/NMDA Receptor Postsynaptic Complex
Authors: Jo Kiwon , Derin Rachel , Li Min , Bredt David S.
Published: April 2018

Protein assembly at the postsynaptic density (PSD) of neuronal synapses is mediated in part by protein interactions with PSD-95/discs large/zona occludens-1 (PDZ) motifs. Here, we identify MALS-1, -2, -3, a family of small synaptic proteins containing little more than a single PDZ domain. MALS-1, -2, and -3 are mammalian homologs LIN-7, aCaenorhabditis elegansprotein essential for vulval development. In contrast to functions for LIN-7 in epithelial cells, MALS-1 and -2 are selectively expressed in specific neuronal populations in brain and are enriched in PSD fractions. In cultured hippocampal neurons, MALS proteins are clustered together with PSD-95 and NMDA type glutamate receptors, consistent with a postsynaptic localization for MALS proteins. Immunoprecipitation and affinity chromatography studies readily identify association of MALS with PSD-95 and an NMDA receptor subunit. The PDZ domain of MALS selectively binds to peptides terminating in E-T/S-R/X-V/I/L, which corresponds to the C terminus of NMDA type 2 receptors and numerous other ion channels at the PSD. This work suggests a role for MALS proteins in regulating recruitment of neurotransmitter receptors to the PSD.