Arc Induction Research Articles

Identification of genes co‐upregulated with Arc during BDNF‐induced long‐term potentiation in adult rat dentate gyrus in vivo

Brain-derived neurotrophic factor (BDNF) is a critical regulator of transcription-dependent adaptive neuronal responses, such as long-term potentiation (LTP). Brief infusion of BDNF into the dentate gyrus of adult anesthetized rats triggers stable LTP at medial perforant path-granule synapses that is transcription-dependent and requires induction of the immediate early gene Arc. Rather than acting alone, Arc is likely to be part of a larger BDNF-induced transcriptional program. Here, we used cDNA microarray expression profiling to search for genes co-upregulated with Arc 3 h after BDNF-LTP induction. Of nine cDNAs encoding for known genes and up-regulated more than four-fold, we selected five genes, Narp, neuritin, ADP-ribosylation factor-like protein-4 (ARL4L), TGF-beta-induced immediate early gene-1 (TIEG1) and CARP, for further validation. Real-time PCR confirmed robust up-regulation of these genes in an independent set of BDNF-LTP experiments, whereas infusion of the control protein cytochrome C had no effect. In situ hybridization histochemistry further revealed up-regulation of all five genes in somata of post-synaptic granule cells following both BDNF-LTP and high-frequency stimulation-induced LTP. While Arc synthesis is critical for local actin polymerization and stable LTP formation, several of the co-upregulated genes have known functions in excitatory synaptogenesis, axon guidance and glutamate receptor clustering. These results provide novel insight into gene expression responses underlying BDNF-induced synaptic consolidation in the adult brain in vivo.

Fornix Lesions Decouple the Induction of Hippocampal Arc Transcription from Behavior But Not Plasticity

The immediate-early gene (IEG) Arc is transcribed after behavioral and physiological treatments that induce synaptic plasticity and is implicated in memory consolidation. The relative contributions of neuronal activity and learning-related plasticity to the behavioral induction of Arc remain to be defined. To differentiate the contributions of each, we assessed the induction of Arc transcription in rats with fornix lesions that impair hippocampal learning yet leave cortical connectivity and neuronal firing essentially intact. Arc expression was assessed after exploration of novel environments and performance of a novel water maze task during which normal rats learned the spatial location of an escape platform. During the same task, rats with fornix lesions learned to approach a visible platform but did not learn its spatial location. Rats with fornix lesions had normal baseline levels of hippocampal Arc mRNA, but unlike normal rats, expression was not increased in response to water maze training. The integrity of signaling pathways controlling Arc expression was demonstrated by stimulation of the medial perforant path, which induced normal synaptic potentiation and Arc in rats with fornix lesions. Together, the results demonstrate that Arc induction can be decoupled from behavior and is more likely to indicate the engagement of synaptic plasticity mechanisms than synaptic or neuronal activity per se. The results further imply that fornix lesions may impair memory in part by decoupling neuronal activity from signaling pathways required for long-lasting hippocampal synaptic plasticity.

Fyn Kinase Induces Synaptic and Cognitive Impairments in a Transgenic Mouse Model of Alzheimer's Disease

Human amyloid precursor protein (hAPP) transgenic mice with high levels of amyloid-beta (Abeta) develop behavioral deficits that correlate with the depletion of synaptic activity-related proteins in the dentate gyrus. The tyrosine kinase Fyn is altered in Alzheimer's disease brains and modulates premature mortality and synaptotoxicity in hAPP mice. To determine whether Fyn also modulates Abeta-induced behavioral deficits and depletions of synaptic activity-dependent proteins, we overexpressed Fyn in neurons of hAPP mice with moderate levels of Abeta production. Compared with nontransgenic controls and singly transgenic mice expressing hAPP or FYN alone, doubly transgenic FYN/hAPP mice had striking depletions of calbindin, Fos, and phosphorylated ERK (extracellular signal-regulated kinase), impaired neuronal induction of Arc, and impaired spatial memory retention. These deficits were qualitatively and quantitatively similar to those otherwise seen only in hAPP mice with higher Abeta levels. Surprisingly, levels of active Fyn were lower in high expresser hAPP mice than in NTG controls and lower in FYN/hAPP mice than in FYN mice. Suppression of Fyn activity may result from dephosphorylation by striatal-enriched phosphatase, which was upregulated in FYN/hAPP mice and in hAPP mice with high levels of Abeta. Thus, increased Fyn expression is sufficient to trigger prominent neuronal deficits in the context of even relatively moderate Abeta levels, and inhibition of Fyn activity may help counteract Abeta-induced impairments.

Molecular reactivity of mesocorticolimbic brain areas of high and low grooming rats after elevated plus maze exposure

High and low grooming rats (HG and LG), selected by extremities in stress-induced self-grooming on the elevated plus maze (EPM), display differences in stress coping style on the EPM, their motivation to self-administer cocaine, and differences in the reactivity of dopaminergic nerve terminals in mesocorticolimbic brain areas. This indicates a link between coping with a stressful/anxiogenic situation and drug intake. Here, we aimed to determine the molecular correlates of these differences by analyzing the reactivity of the mesocorticolimbic brain areas (the medial prefrontal cortex (mPFC) nucleus accumbens shell (NAS) and ventral tegmental area (VTA)) of HG and LG rats in response to EPM exposure. We report by measuring levels of immediate early gene (IEG) transcripts that EPM exposure-induced IEG expression was not significantly different between HG and LG rats. On the other hand, novel IEG expression patterns upon stress (EPM exposure) were apparent in all three areas including arc induction in the mPFC and NAS, CRH, BDNF, and Nr4a3 induction in the NAS, and serum glucocorticoid-regulated kinase (sgk) induction in the VTA. It is concluded that although the mPFC, NAS, and VTA play a role in modulating stress and grooming behavior, the neuronal reactivity in these regions measured by the IEG response is not related with behavioral extremities in stress coping style displayed on the EPM.

Multiple periods of functional ocular dominance plasticity in mouse visual cortex

The precise period when experience shapes neural circuits in the mouse visual system is unknown. We used Arc induction to monitor the functional pattern of ipsilateral eye representation in cortex during normal development and after visual deprivation. After monocular deprivation during the critical period, Arc induction reflects ocular dominance (OD) shifts within the binocular zone. Arc induction also reports faithfully expected OD shifts in cat. Shifts towards the open eye and weakening of the deprived eye were seen in layer 4 after the critical period ends and also before it begins. These shifts include an unexpected spatial expansion of Arc induction into the monocular zone. However, this plasticity is not present in adult layer 6. Thus, functionally assessed OD can be altered in cortex by ocular imbalances substantially earlier and far later than expected.

Morphine activates Arc expression in the mouse striatum and in mouse neuroblastoma Neuro2A MOR1A cells expressing μ-opioid receptors

Activity-regulated cytoskeleton-associated protein (Arc) is an effector immediate early gene product implicated in long-term potentiation and other forms of neuroplasticity. Earlier studies demonstrated Arc induction in discrete brain regions by several psychoactive substances, including drugs of abuse. In the present experiments, the influence of morphine on Arc expression was assessed by quantitative reverse transcription real-time PCR and Western blotting in vivo in the mouse striatum/nucleus accumbens and, in vitro, in the mouse Neuro2A MOR1A cell line, expressing mu-opioid receptor. An acute administration of morphine produced a marked increase in Arc mRNA and protein level in the mouse striatum/nucleus accumbens complex. After prolonged opiate treatment, tolerance to the stimulatory effect of morphine on Arc expression developed. No changes in the striatal Arc mRNA levels were observed during spontaneous or opioid antagonist-precipitated morphine withdrawal. In Neuro2A MOR1A cells, acute, but not prolonged, morphine treatment elevated Arc mRNA level by activation of mu-opioid receptor. This was accompanied by a corresponding increase in Arc protein level. Inhibition experiments revealed that morphine induced Arc expression in Neuro2A MOR1A cells via intracellular signaling pathways involving mitogen-activated protein (MAP) kinases and protein kinase C. These results lend further support to the notion that stimulation of opioid receptors may exert an activating influence on some intracellular pathways and leads to induction of immediate early genes. They also demonstrate that Arc is induced in the brain in vivo after morphine administration and thus may play a role in neuroadaptations produced by the drug.

Synaptic Targeting by Alzheimer's-Related Amyloid β Oligomers

The cognitive hallmark of early Alzheimer's disease (AD) is an extraordinary inability to form new memories. For many years, this dementia was attributed to nerve-cell death induced by deposits of fibrillar amyloid beta (Abeta). A newer hypothesis has emerged, however, in which early memory loss is considered a synapse failure caused by soluble Abeta oligomers. Such oligomers rapidly block long-term potentiation, a classic experimental paradigm for synaptic plasticity, and they are strikingly elevated in AD brain tissue and transgenic-mouse AD models. The current work characterizes the manner in which Abeta oligomers attack neurons. Antibodies raised against synthetic oligomers applied to AD brain sections were found to give diffuse stain around neuronal cell bodies, suggestive of a dendritic pattern, whereas soluble brain extracts showed robust AD-dependent reactivity in dot immunoblots. Antigens in unfractionated AD extracts attached with specificity to cultured rat hippocampal neurons, binding within dendritic arbors at discrete puncta. Crude fractionation showed ligand size to be between 10 and 100 kDa. Synthetic Abeta oligomers of the same size gave identical punctate binding, which was highly selective for particular neurons. Image analysis by confocal double-label immunofluorescence established that >90% of the punctate oligomer binding sites colocalized with the synaptic marker PSD-95 (postsynaptic density protein 95). Synaptic binding was accompanied by ectopic induction of Arc, a synaptic immediate-early gene, the overexpression of which has been linked to dysfunctional learning. Results suggest the hypothesis that targeting and functional disruption of particular synapses by Abeta oligomers may provide a molecular basis for the specific loss of memory function in early AD.

Stress-induced activation of the immediate early gene Arc (activity-regulated cytoskeleton-associated protein) is restricted to telencephalic areas in the rat brain: relationship to c-fos mRNA.

Arc is an effector immediate early gene whose expression is induced in situations of increased neuronal activity. However, there is no report on the influence of stress on Arc expression. Here, we compared the induction of both c-fos and Arc mRNAs in the brain of rats exposed to one of three different stressful situations: novel environment, forced swimming and immobilization. An absent or weak c-fos mRNA signal was observed in control rats, whereas those exposed to one of three stressors showed enhanced c-fos expression in a wide range of brain areas. Constitutive Arc expression was observed in some areas such as cortex, striatum, hippocampus, reticular thalamic nucleus and cerebellar cortex. In response to stressors, a strong induction of Arc was observed, but the pattern was different from that of c-fos. For instance, activation of Arc but not c-fos was observed in the nucleus accumbens after immobilization and in the hippocampus after novel environment. No Arc induction was observed in diencephalic and brainstem areas. The present data show that Arc has a neuroanatomically restricted pattern of induction in the brain after emotional stress. Telencephalic activation suggests that a more intense induction of synaptic plasticity is occurring in this area after exposure to emotional stressors.

Arc interacts with microtubules/microtubule-associated protein 2 and attenuates microtubule-associated protein 2 immunoreactivity in the dendrites.

Arc, activity-regulated cytoskeleton-associated gene, is an immediate early gene, and its expression is regulated by a variety of stimuli, such as electric stimulation and methamphetamine. The function of Arc, however, is unknown. To explore this function, we carried out expression experiments by transfecting green fluorescent protein (GFP)-Arc constructs or by using a protein transduction system in hippocampal cultured neurons. We found that the overexpression of Arc as well as Arc induction by seizure in vivo decreased microtubule-associated protein 2 (MAP2) staining in the dendrites by immunocytochemistry, although MAP2 content was not changed on Western blot. Furthermore, Arc interacted with newly polymerized microtubules and MAP2, leading to blocking of the epitope of MAP2. The data suggest that Arc increased by synaptic activities would trigger dendritic remodeling by interacting with cytoskeletal proteins.

Muscarinic acetylcholine receptor stimulation induces expression of the activity-regulated cytoskeleton-associated gene (ARC)

Muscarinic acetylcholine receptors (mAChR) are involved in learning and memory but their molecular function in these processes is not fully understood. In this study, the signal transduction pathway coupling mAChR activation to induction of the activity-regulated cytoskeleton-associated gene (ARC) was examined. ARC was first identified as an effector immediate early gene induced by neuronal activity and ARC protein is thought to play a role in synaptic plasticity. In rats, intraperitoneal injection of pilocarpine, a potent agonist of mAChR, led to increased ARC expression in the brain. In human SH-SY5Y neuroblastoma cells mAChR stimulation with carbachol caused a rapid and robust induction of ARC expression. This effect was inhibited by atropine, a nonselective muscarinic receptor antagonist as well as by M1/M3 subtype-specific antagonists. Analysis of mAChR downstream effectors revealed that protein kinase C (PKC) and tyrosine kinases of the src family are key molecules in the signal cascade leading to ARC expression. Our data suggest, for the first time, that a correlation exists among mAChR-controlled signal cascades, the induction of the effector immediate early gene ARC and synaptic plasticity.

Rapid induction of Arc is observed in the granule cell dendrites in the accessory olfactory bulb after mating

The activity-regulated cytoskeleton-associated protein (Arc), encoded by the immediate early gene arc, is enriched in the brain and is hypothesized to play a role in the activity-dependent neuronal plasticity in the hippocampus. In the present study, the time course of Arc expression during the post-mating period was determined immunocytochemically, and the localization of Arc in the neurons in the accessory olfactory bulb (AOB) of female mice after mating was analyzed using immunocytochemical electron microscopy. Transient increases in the number of Arc-immunoreactive cells were observed in the glomerular, mitral/tufted cell and granule cell layers of the AOB after mating. In particular, the increase in the granule cell layer was remarkable, and larger than the increases in the other layers. In addition, electron microscopic observation revealed that Arc immunoreactivity was in the dendrites of the granule cells 1.5 h after mating. These results indicate that expression of Arc protein is induced rapidly and transiently in granule cell dendrites after mating. It is postulated that Arc protein has a role in the neuronal plasticity of the AOB after mating.

Local Synthesis of Proteins at Synaptic Sites on Dendrites: Role in Synaptic Plasticity and Memory Consolidation?

Long-lasting forms of memory are thought to be mediated by modifications in synapses that are induced by particular patterns of activity, take time to be established (consolidated), and involve new gene expression. The molecular mechanisms underlying such long-lasting synaptic modifications remain to be defined. Here, we focus on new findings regarding synapse-specific gene expression and especially the intracellular transport and synaptic targeting of the mRNA for a recently identified immediate early gene called activity-regulated cytoskeleton-associated protein ( Arc). Arc mRNA is transported into dendrites after episodes of neuronal activation. The newly synthesized Arc mRNA localizes selectively at synapses that have experienced particular patterns of activity, and this targeting depends on NMDA receptor activation. Arc protein is assembled into the postsynaptic junction, and Arc expression is strongly induced by brief behavioral experiences in select populations of neurons. These features of Arc induction and trafficking reveal novel cellular mechanisms that are well suited to mediate long-term synapse-specific modifications. We will review findings from other laboratories that both LTP and memory are disrupted when Arc induction is blocked, and we will discuss the possibility that Arc protein may play a key role in activity-dependent synaptic modification.

Magnetic properties of the low-temperature phase of MnBi

MnBi forms peritectically at ∼450 °C. Preparation of MnBi employing conventional techniques such as arc melting and induction melting results in the segregation of manganese. In order to avoid this segregation, we followed the procedure recommended by Guo et al. [X. Guo, A. Zaluska, Z. Altounian, and J. O. Strom-Olsen, J. Mater. Res. 5, 2646 (1990)] and prepared a low-temperature phase of MnBi by melt spinning, followed by heat treatment. Fine powder of MnBi was prepared by ball milling the melt-spun ribbons for various lengths of time. Magnetic properties of these powders were determined. In particular, the temperature dependent coercivity was studied from room temperature to 360 °C for the powders ball milled for 2 and 10 h. The coercivity is found to increase with the increase in temperature reaching a maximum of 25.8 kOe at 280 °C and then decrease as the temperature is increased further. We also found that a peak in coercivity is observed for the samples milled for 10 h. MnBi shows a first-order transition to a paramagnetic phase at 360 °C. In an attempt to increase this transition temperature, an alloy of composition Mn0.75Ni0.25Bi0.5Sb0.5 was made by induction melting. The transition temperature increases from 360 °C for MnBi to 400 °C for Mn0.75Ni0.25Bi0.5Sb0.5.

Cold crucible levitation melting of biomedical Ti-30 wt%Ta alloy.

Recently, titanium-tantalum alloys have been studied as implant materials for dental and orthopedic surgery. However, titanium and tantalum are difficult to mix by common arc melting and induction melting, because of their high melting point and the marked difference between their densities (Ti: 1,680 degrees C, 4.5 g/cm3, Ta: 2,990 degrees C, 16.6 g/cm3). Thus, the Cold Crucible Levitation Melting (CCLM) method was chosen to produce a Ti-30 wt%Ta binary alloy in the present study. The CCLM furnace, with 1 kg capacity, consisted of a water-cooled crucible comprising oxygen-free high purity copper segments and coils wrapped around the crucible and connected to a frequency inverter power supply. A qualified ingot of 1.0 kg of Ti-30 wt%Ta alloy was obtained. The ingot was characterized from the surface quality, chemical composition distribution and microstructure, and finally the melting process was discussed.

Breaking up two-layer cast-ion rolling-mill rolls with a chill layer

When the Magnitogorsk Metallurgical Combine began to make cast-iron rolls with a chill layer, it became necessary to break up the rolls after they had been removed from service. The rolls had to be divided into pieces no larger than 600 • 800 mm in order to be used as a component of the metallic charge for the production of alloyed cast iron in arc furnaces or induction furnaces. The combine conducted an experiment in which the rolls were broken up by two technologies already in use at the combine explosive fragmentation, with the blastholes for the explosive charges being perpendicular to the axis of the roll; breakup of the rolls on an impact breaker. The first method posed the danger of having fragments of the chill layer fly off as the blastholes were being drilled. In the second method, the body of the roll could not be broken up into pieces of the required dimensions, and the ball-weight of the impact breaker would itself be broken due to the high hardness of the body. Thus, two new technologies were proposed and tried out for breaking up two-layer cast-iron rolls explosive fragmentation in which the blastholes are located along the body of the roll; the use of an impact breaker after the roll body has been heated by a powerful mechanical cutter in the area where the roll is supposed to be divided. Explosive fragmentation with axial blastholes was chosen as the main technology after tests (two opposing blastholes are used). To perform a comparative analysis, nine rolls were broken up by the proposed fragmentation technology (A) and one was broken up by the existing technology (B). The results (in terms of one roll) are shown below:

The activity-regulated cytoskeletal-associated protein arc is expressed in different striosome-matrix patterns following exposure to amphetamine and cocaine.

The activity-regulated, cytoskeletal-associated gene, arc, is a brain-enriched immediate-early gene whose expression is rapidly induced in the striatum by dopamine receptor agonists. This rapid induction of arc in the striatum is similar to that of other early response genes such as c-fos, junB, deltafosB, fra, and NGFI-A, which code for transcription factors. Unlike these proteins, however, Arc is a cytoskeletal protein expressed not only in the nucleus of neurons but also in their dendrites. We investigated the patterns of Arc expression evoked in the rat striatum by acute exposures to two psychomotor stimulants, cocaine and amphetamine. Cocaine induced arc in striatal neurons that were broadly distributed within both striosome and matrix compartments of the caudoputamen. Amphetamine also evoked Arc expression in striatal projection neurons, but these were heavily concentrated in the striosomal compartment and only sparsely in the matrix compartment in the rostral striatum. The contrasting patterns of Arc expression evoked by cocaine and amphetamine parallel those of c-Fos, JunB, FRA, and NGFI-A expression induced by these two psychomotor stimulants. This difference in the action of cocaine and amphetamine at the level of protein expression may be linked to the different effects of these psychomotor stimulants on behavior.

The inductance of the discharge in a spark gap

The inductance of an arc discharge has been modeled and a mathematical formula given for the arc inductance in terms of its geometrical dimensions. This formula agrees with the experimental results and is used to determine the arc-channel radius. Inductance measurements have been taken for various values of pressure, interelectrode distance and applied voltage in a triggered spark gap. The dependence of inductance upon these parameters has been explained through the existing variations of the arc-channel cross section. Two mechanisms are responsible for the variations of inductance. The first is diffusion and the second is the tendency of the current channel to vary diameter with pressure, interelectrode distance, and applied voltage. The time history of the arc-channel inductance has been investigated. Finally, the ambipolar diffusion coefficient and the total charge number of the arc channel has been obtained.

Overdamped arc discharge data and an AWA model

Experimental data on overdamped arc discharges are compared with a lumped-circuit discharge mode employing the arc welder's ansatz (AWA). The AWA prescribes that the arc resistance-varies time as R/sub a/(t)=V*/ mod I(t) mod , where V* is a positive constant and I(t) is the discharge current. In the circuit, in addition to the time-dependent arc resistance R/sub a/ and a small arc inductance L/sub a/, there are an external time-dependent series resistance R/sub o/, inductance L/sub o/, and source capacitance C, the values of which are given. The AWA theory is compared with an observed arc current pulse I(t) by using these given values and normalizing the theory to one point on the data curve; an adequate fit to the data is obtained. Data for the dynamic arc resistance R/sub a/(t) are also compared with the AWA prescription and with other available theories for R/sub a/, which also use a one-point normalization. The AWA form for R/sub a/ compares favorably with the other theories.< <ETX xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">&gt;</ETX>

ChemInform Abstract: Crystallographic and Magnetic Properties of a New Series RFe10SiC0.5 (R: Ce, Pr, Nd, Sm).

AbstractThe title compounds are prepared by arc melting and induction techniques starting from stoichiometric amounts of the binary compounds.

Hydrogen production and storage using titanium electrodes and metal hydrides

The development of a cheap, safe method for the production and storage of hydrogen gas could lead to the more widespread use of hydrogen as a fuel, particularly for motor vehicles. The aims of this project are to improve the efficiency of hydrogen production by the solar-assisted electrolysis of water, and to improve the storage processes, in particular those using hydrides, with a view to designing, building and demonstrating a prototype system at a later stage. Titanium electrodes were developed for producing hydrogen from solar energy, at an efficiency of approximately 2%, and an Australian patent was obtained for the process. Further improvements in efficiency were sought by preparing other semi-conducting films, modifying the titanium electrodes, and investigating the mechanisms of hydrogen production. The spectral response of titanium oxide electrodes was broadened slightly by chromium, but the overall efficiency decreased. Iron/iron oxide and several metal/metal sulphide and selenide systems appear promising. Hydrogen was stored as iron-titanium hydride using alloys prepared by argon arc melting and induction melting in batches of up to 2 kg weight. Facilities are available for 100 kg batches. The alloy structures and properties were evaluated and compared with those of commercially available alloys. Some collaborative work was commenced with Melbourne University on a hydrogen-fuelled vehicle which will use iron-titanium hydride.