Local Microinfusion Research Articles

O003 / #714 COGNITIVE RESTORATION IN ALZHEIMER’S PATIENTS BY LOCAL INTRACEREBRAL CELL-MEDIATED NERVE GROWTH FACTOR DELIVERY: CLINICAL EFFICACY AND FURTHER METHOD OPTIMIZATION: ORAL POSTER PRESENTATIONS FIVE BEST ABSTRACTS

Local neuromodulation of brain functions can be accomplished in many ways: electrical stimulation, local micro-infusion of drugs, implantations of viruses or engineered cells, etc. We have studied targeted encapsulated cell implants delivering active substances locally in Alzheimer patients (AD). AD is associated with early loss of basal forebrain cholinergic neurons (BFCNs) and cognitive dysfunction. BFCNs are dependent on nerve growth factor (NGF). Previously, we have developed an encapsulated cell biodelivery (ECB) platform and utilized it to stimulate BFCNs in two cholinergic target sites (Ch4 [nucl.

In Vivo Detection of Redox-Inactive Neurochemicals in the Rat Brain with an Ion Transfer Microsensor.

Electrochemical tracking of redox-inactive neurochemicals remain a challenge due to chemical inertness, almost no Faraday electron transfer for these species, and the complex brain atmosphere. In this work, we demonstrate a low-cost, simple-making liquid/liquid interface microsensor (LLIM) to monitor redox-inactive neurochemicals in the rat brain. Taking choline (Ch) as an example, based on the difference in solvation energies of Ch in cerebrospinal fluid (aqueous phase) and 1,2-dichloroethane (1,2-DCE; organic phase), Ch is recognized in the specific ion-transfer potential and distinctive ion-transfer current signals. The LLIM has an excellent response to Ch with good linearity and selectivity, and the detection limit is 0.37 μM. The LLIM can monitor the dynamics of Ch in the cortex of the rat brain by both local microinfusion and intraperitoneal injection of Ch. This work first demonstrates that the LLIM can be successfully applied in the brain and obtain electrochemical signals in such a sophisticated system, allowing one new perspective of sensing at the liquid/liquid interface for nonelectrically active substances in vivo to understand the physiological function of the brain.

Ag2S/Ag Nanoparticle Microelectrodes for In Vivo Potentiometric Measurement of Hydrogen Sulfide Dynamics in the Rat Brain

Hydrogen sulfide (H2S) plays a pivotal role in gas signal transduction, neuroprotection, and regulation of physiological and pathological processes. However, in vivo tracking the dynamic of hydrogen sulfide in the complex brain environment still faces huge challenges. This study demonstrates a new potentiometric method to monitor in vivo the dynamics of hydrogen sulfide in the rat brain using silver nanoparticles (AgNPs)-modified carbon fiber microelectrodes (AgNPs/CFE) pretreated with Na2S (i.e., Ag2S/AgNPs/CFE), which acts as a solid-contact and ion-selective microelectrode. The Ag2S/AgNPs/CFE exhibits good potential response toward hydrogen sulfide in the range of 2.5-160 μM, with a detection limit of 0.8 μM. Because of the presence of Ag2S, the Ag2S/AgNPs/CFE shows good selectivity to hydrogen sulfide, avoiding the interference from coexistent electroactive neurochemicals and the analogies, such as ascorbic acid and cysteine in the central nervous system. This good selectivity combined with the reversibility, protein antifouling, and biocompatibility of the microelectrode enables the Ag2S/AgNPs/CFE to detect hydrogen sulfide in the rat brain during local microinfusion of Na2S and the change in pH. Our study provides a reliable method to track hydrogen sulfide selectively in vivo, which will help to explore the function of hydrogen sulfide in neurophysiology and pathology.

Frequency- and state-dependent effects of hippocampal neural disinhibition on hippocampal local field potential oscillations in anesthetized rats.

Reduced inhibitory GABA function, so-called neural disinhibition, has been implicated in cognitive disorders, including schizophrenia and age-related cognitive decline. We previously showed in rats that hippocampal disinhibition by local microinfusion of the GABA-A receptor antagonist picrotoxin disrupted memory and attention and enhanced hippocampal multi-unit burst firing recorded around the infusion site under isoflurane anesthesia. Here, we analyzed the hippocampal local field potential (LFP) recorded alongside the multi-unit data. We predicted frequency-specific LFP changes, based on previous studies implicating GABA in hippocampal oscillations, with the weight of evidence suggesting that disinhibition would facilitate theta and disrupt gamma oscillations. Using a new semi-automated method based on the kurtosis of the LFP peak-amplitude distribution as well as on amplitude envelope thresholding, we separated three distinct hippocampal LFP states under isoflurane anesthesia: "burst" and "suppression" states-high-amplitude LFP spike bursts and the interspersed low-amplitudeperiods-and a medium-amplitude "continuous" state. The burst state showed greater overall power than suppression and continuous states and higher relative delta/theta power, but lower relative beta/gamma power. The burst state also showed reduced functional connectivity across the hippocampal recording area, especially around theta and beta frequencies. Overall neuronal firing was higher in the burst than the other two states, whereas the proportion of burst firing was higher in burst and continuous states than the suppression state. Disinhibition caused state- and frequency-dependent LFP changes, tending to increase power at lower frequencies (<20 Hz), but to decrease power and connectivity at higher frequencies (>20 Hz) in burst and suppression states. The disinhibition-induced enhancement of multi-unit bursting was also state-dependent, tending to be more pronounced in burst and suppression states than the continuous state. Overall, we characterized three distinct hippocampal LFP states in isoflurane-anesthetized rats. Disinhibition changed hippocampal LFP oscillations in a state- and frequency-dependent way. Moreover, the disinhibition-induced enhancement of multi-unit bursting was also LFP state-dependent.

Cannabinoid agonist administration within the cerebellar cortex impairs motor learning.

Systemic administration of cannabinoid agonists impairs cerebellum-dependent motor learning. The cannabinoid-induced impairment of motor learning has been hypothesized to be due to disruption of Purkinje cell plasticity within the cerebellar cortex. In the current study, we tested this hypothesis in rats with localized microinfusions of cannabinoid agonists and antagonists into the cerebellar cortex during eyeblink conditioning, a type of cerebellum-dependent motor learning. Infusions of the cannabinoid agonists WIN55,212-2 or ACEA directly into the eyeblink conditioning microzone of the cerebellar cortex severely impaired acquisition of eyeblink conditioning, whereas the CB1R antagonist SR141716A did not produce a significant impairment. Infusions of WIN55,212-2 outside of the eyeblink conditioning microzone did not impair motor learning, establishing anatomical specificity for the agonist effects. The motor learning impairment caused by WIN55,212-2 and ACEA was rescued by SR141716A, indicating that the learning deficit was produced through CB1Rs. The current findings demonstrate that the effects of cannabinoid receptor agonists on motor learning are localized to CB1Rs within a discrete microzone of the cerebellar cortex.

Effects of nucleus accumbens oxytocin and its antagonist on social approach behavior.

Severe impairment of social interaction is a core symptom of numerous psychiatric disorders. Oxytocin (OT) has been shown to be involved in various aspects of social behavior related to reproduction, but little is known about its effects on nonreproductive social interaction between adults or the neuroanatomical location where OT exerts its action. Here, we examined the nucleus accumbens, a region of the brain containing high levels of the oxytocin receptor (OTR) and comprising an important node in the neural circuitry possibly related to social interaction. Behavioral effects of a local microinfusion of OT (0.1, 1, and 10 ng/side) and an oxytocin receptor antagonist (OTR-A) (1, 10, and 100 ng/side) were evaluated in naturally high social and low social female and male monogamous mandarin voles (Microtus mandarinus) using the social preference paradigm and open-field tests. The results showed that administration of 1 ng/side OT increased social preference; however, this effect was not apparent at lower or higher doses. OT did not alter anxiety-like behavior or total locomotion. Microinfusions of a selective OTR-A at 10 and 100 ng doses reduced social approach behavior; a dose of 1 ng had no effect. In conclusion, our results suggest that accumbal OT and OTR-A regulate social preferences in voles in a dose-dependent manner.

Dopamine D1 receptor activity is involved in the increased anxiety levels observed in STZ-induced diabetes in rats

Epidemiological surveys have indicated that anxiety disorders are more frequent in diabetic patients than in the general population. Similar results have been shown in animal studies using the streptozotocin (STZ)-induced diabetes model. The mechanisms underlying this relationship are not clearly understood, but it has been suggested that alterations in the dopaminergic neurotransmission, which plays an important role in the amygdaloid modulation of fear and anxiety, may be involved. The aim of this study was to ascertain whether or not the amygdaloid DA D1 receptors are involved in the increase of anxiety-like behavior observed in “diabetic” animals. Adult Wistar male rats were injected with STZ (50mg/kg, i.p.) in two consecutive days and subjected to the Shock-Probe Burying Test 10days after the beginning of treatment. STZ-treated rats showed a significant increase in immobility/freezing behavior whereas no effects were elicited in latency to bury, burying behavior itself and the number of shocks received during testing as compared with non-diabetic controls. These results suggest the triggering of a passive coping response in the STZ-treated rats. Interestingly, immobility/freezing behavior was reversed following the intra-amygdaloid dopamine D1 receptor blockade by the local microinfusion of SCH23390 (100ng/side). Autoradiographic experiments showed a selective increase of [3H]-SCH23390 binding in the ventral intercalated paracapsular islands of STZ-treated rats when compared to the non-treated control group. Our results suggest that a hyperdopaminergic state involving DA D1 receptors within the amygdala may have a role in the increase of anxiety observed in diabetic rats.

Temporary inactivation reveals that the CA1 region of the mouse dorsal hippocampus plays an equivalent role in the retrieval of long-term object memory and spatial memory

Recognition of a previously experienced item or object depends upon the successful retrieval of memory for the object. The neural mechanisms that support object recognition memory in the mammalian brain are not well understood. The rodent hippocampus plays a well-established role in spatial memory, and we previously demonstrated that temporary inactivation of the mouse hippocampus impairs object memory, as assessed with a novel object preference (NOP) test. The present studies were designed to test some remaining issues regarding the contribution of the CA1 sub-region of the mouse dorsal hippocampus to long-term object memory. Specifically, we examined whether the retrieval of spatial memory (as assessed by the Morris water maze; MWM) and object recognition memory are differentially sensitive to inactivation of the CA1 region. The current study used pre-test local microinfusion of muscimol directly into the CA1 region of dorsal hippocampus to temporarily interrupt its function during the respective retrieval phases of both behavioral tasks, in order to compare the contribution of the CA1 to object memory and spatial memory. Histological analyses revealed that local intra-CA1 injection of muscimol diffused within, and not beyond, the CA1 region of dorsal hippocampus. The degree of memory retrieval impairment induced by muscimol was comparable in the two tasks, supporting the view that object memory and spatial memory depend similarly on the CA1 region of rodent hippocampus. Further, we confirmed that the muscimol-induced impairment of CA1 function is temporary. First, mice that exhibited impaired object memory retrieval immediately after intra-CA1 muscimol, subsequently exhibited unimpaired retrieval of object memory when tested 24h later. Secondly, a cohort of mice that exhibited impaired object memory retrieval after intra-CA1 muscimol later acquired spatial memory in the MWM comparable to that of control mice. Together, these results offer further support for the involvement of the CA1 region of mouse hippocampus in object recognition memory, and provide evidence to suggest that the NOP task is as much a test of hippocampal function as the classic MWM test.

Learning history and cholinergic modulation in the dorsal hippocampus are necessary for rats to infer the status of a hidden event.

Identifying statistical patterns between environmental stimuli enables organisms to respond adaptively when cues are later observed. However, stimuli are often obscured from detection, necessitating behavior under conditions of ambiguity. Considerable evidence indicates decisions under ambiguity rely on inference processes that draw on past experiences to generate predictions under novel conditions. Despite the high demand for this process and the observation that it deteriorates disproportionately with age, the underlying mechanisms remain unknown. We developed a rodent model of decision-making during ambiguity to examine features of experience that contribute to inference. Rats learned either a simple (positive patterning) or complex (negative patterning) instrumental discrimination between the illumination of one or two lights. During test, only one light was lit while the other relevant light was blocked from physical detection (covered by an opaque shield, rendering its status ambiguous). We found experience with the complex negative patterning discrimination was necessary for rats to behave sensitively to the ambiguous test situation. These rats behaved as if they inferred the presence of the hidden light, responding differently than when the light was explicitly absent (uncovered and unlit). Differential expression profiles of the immediate early gene cFos indicated hippocampal involvement in the inference process while localized microinfusions of the muscarinic antagonist, scopolamine, into the dorsal hippocampus caused rats to behave as if only one light was present. That is, blocking cholinergic modulation prevented the rat from inferring the presence of the hidden light. Collectively, these results suggest cholinergic modulation mediates recruitment of hippocampal processes related to past experiences and transfer of these processes to make decisions during ambiguous situations. Our results correspond with correlations observed between human brain function and inference abilities, suggesting our experiments may inform interventions to alleviate or prevent cognitive dysfunction. © 2015 Wiley Periodicals, Inc.

Nicotine receptors mediating sensorimotor gating and its enhancement by systemic nicotine.

Prepulse inhibition (PPI) of startle occurs when intensity stimuli precede stronger startle-inducing stimuli by 10–1000 ms. PPI deficits are found in individuals with schizophrenia and other psychiatric disorders, and they correlate with other cognitive impairments. Animal research and clinical studies have demonstrated that both PPI and cognitive function can be enhanced by nicotine. PPI has been shown to be mediated, at least in part, by mesopontine cholinergic neurons that project to pontine startle neurons and activate muscarinic and potentially nicotine receptors (nAChRs). The subtypes and anatomical location of nAChRs involved in mediating and modulating PPI remain unresolved. We tested the hypothesis that nAChRs that are expressed by pontine startle neurons contribute to PPI. We also explored whether or not these pontine receptors are responsible for the nicotine enhancement of PPI. While systemic administration of nAChR antagonists had limited effects on PPI, PnC microinfusions of the non-α7nAChR preferring antagonist TMPH, but not of the α7nAChR antagonist MLA, into the PnC significantly reduced PPI. Electrophysiological recordings from startle-mediating PnC neurons confirmed that nicotine affects excitability of PnC neurons, which could be antagonized by TMPH, but not by MLA, indicating the expression of non-α7nAChR. In contrast, systemic nicotine enhancement of PPI was only reversed by systemic MLA and not by TMPH or local microinfusions of MLA into the PnC. In summary, our data indicate that non-α7nAChRs in the PnC contribute to PPI at stimulus intervals of 100 ms or less, whereas activation of α7nAChRs in other brain areas is responsible for the systemic nicotine enhancement of PPI. This is important knowledge for the correct interpretation of behavioral, preclinical, and clinical data as well as for developing drugs for the amelioration of PPI deficits and the enhancement of cognitive function.

Oxytocin microinjected into the central amygdaloid nuclei exerts anti-aggressive effects in male rats

We recently demonstrated that acute and chronic intracerebroventricular enhancement of brain OXT levels induces potent anti-aggressive and pro-social explorative effects during social challenges. However, the exact anatomical location in the brain where OXT exerts its action is still elusive. In the present study, we targeted two critical brain areas, i.e. the central amygdala (CeA) and the dorsal raphe (DR), both containing high levels of OXT receptors (OXTRs) and constituting important nodes of the neural circuitry related to aggression. Behavioral effects of local micro-infusion of OXT and OXTR antagonist, L368.899, (alone and combined) were evaluated in resident male rats during confrontations with an unfamiliar male intruder. Our results show that OXT microinjected into the CeA markedly reduced resident's offensive behavior and facilitated social exploration, without affecting other non-aggressive behaviors. The receptor specificity of the behavioral effects was verified when a micro-infusion of a selective OXTR antagonist nullified the changes. Pharmacological blockade of CeA OXTRs per se was without clear behavioral effects suggesting that endogenous OXT within the CeA does not play a major inhibitory role on offensiveness. Anatomical specificity was also supported by the absence of relevant behavioral effects when OXT was microinjected into more medial sub-regions of the amygdala. Likewise, within the DR neither OXT nor OXTR exerted significant effects on offensive aggression, while microinjection of the 5-HT1A autoreceptor agonist in this region significantly suppressed aggression.In conclusion, our results point at the CeA as an important brain site of action for the anti-aggressive and pro-social explorative effects induced by exogenous enhancement of brain OXT levels.

Genetic or pharmacological reduction of PERK enhances cortical-dependent taste learning.

Protein translation initiation is controlled by levels of eIF2α phosphorylation (p-eIF2α) on Ser51. In addition, increased p-eIF2α levels impair long-term synaptic plasticity and memory consolidation, whereas decreased levels enhance them. Levels of p-eIF2α are determined by four kinases, of which protein kinase RNA-activated (PKR), PKR-like endoplastic reticulum kinase (PERK), and general control nonderepressible 2 are extensively expressed in the mammalian mature brain. Following identification of PERK as the major kinase to determine basal levels of p-eIF2α in primary neuronal cultures, we tested its function as a physiological constraint of memory consolidation in the cortex, the brain structure suggested to store, at least in part, long-term memories in the mammalian brain. To that aim, insular cortex (IC)-dependent positive and negative forms of taste learning were used. Genetic reduction of PERK expression was accomplished by local microinfusion of a lentivirus harboring PERK Short hairpin RNA, and pharmacological inhibition was achieved by local microinfusion of a PERK-specific inhibitor (GSK2606414) to the rat IC. Both genetic reduction of PERK expression and pharmacological inhibition of its activity reduced p-eIF2α levels and enhanced novel taste learning and conditioned taste aversion, but not memory retrieval. Moreover, enhanced extinction was observed together with enhanced associative memory, suggesting increased cortical-dependent behavioral plasticity. The results suggest that, by phosphorylating eIF2α, PERK functions in the cortex as a physiological constraint of memory consolidation, and its downregulation serves as cognitive enhancement.

Noradrenergic influences in the basolateral amygdala on inhibitory avoidance memory are mediated by an action on α2-adrenoceptors.

The role of norepinephrine (NE) in the consolidation of inhibitory avoidance learning (IA) in rats is known to involve α1- and β-adrenoceptor systems in the basolateral nucleus of the amygdala (BLA). However, the amygdala also contains α2-adrenoceptor subtypes, and local microinfusions of the selective α2-adrenoceptor antagonist idazoxan and agonist UK 14,304 respectively into the BLA enhance and inhibit IA performances when administered before acquisition. The present study investigated whether the effects of idazoxan and UK 14,304 on IA were associated with changes in NE release within the BLA before and after one-trial inhibitory avoidance training. Male Sprague-Dawley rats were unilaterally implanted with a microdialysis probe in the BLA and were administered idazoxan (0.1mM) or UK 14,304 (10 μM) by retrodialysis infusion 15 min before the acquisition of IA. Dialysates were collected every 15 min for analysis of NE. Retrodialysis of idazoxan potentiated the release of NE induced by footshock application, whereas UK 14,304 decreased NE release to the extent that the footshock failed to induce any measurable effect on NE levels. Idazoxan infusion enhanced IA retention tested 24h later and this effect was directly related to the level of NE release in the BLA measured during IA acquisition. In contrast, the infusion of UK 14,304 did not modify IA performances in comparison to control animals, possibly due to compensatory activity of the contralateral BLA. These results are consistent with previous evidence that amygdala NE is involved in modulating memory consolidation, and provide evidence for an involvement of presynaptic α2-autoceptors in the BLA in this process.

Involvement of cannabinoid receptors in the amygdala and prefrontal cortex of rats in fear learning, consolidation, retrieval and extinction

Cannabinoid receptors 1 (CB1R) have been shown to be a crucial part of the neuromodulatory endocannabinoid system which is involved in emotional learning and memory.We here investigated in rats the role of CB1R in the basolateral amygdala (BLA) and medial prefrontal cortex (mPFC) in different phases of fear learning, memory and extinction. We used the fear potentiated startle paradigm to measure the effects of local microinfusion of the CB1R agonist WIN 55,212-2 (WIN) or the CB1R antagonist AM251 on acquisition, consolidation, retrieval and extinction of fear.No effects on fear acquisition of WIN or AM251 were found in the BLA or mPFC. WIN impaired fear retrieval in the BLA and in mPFC. Also, WIN reduced fear consolidation in the BLA but not in the mPFC. AM251 decreased fear consolidation after mPFC infusion. Likewise, fear extinction was impaired by AM251 infused into the mPFC.Our data indicate that fear memory consolidation and retrieval, as well as extinction are regulated differentially by amygdaloid and cortical CB1R.

Marked resistance of femoral adipose tissue blood flow and lipolysis to adrenaline in vivo

Fatty acid entrapment in femoral adipose tissue has been proposed to prevent ectopic fat deposition and visceral fat accumulation, resulting in protection from insulin resistance. Our objective was to test the hypothesis of femoral, compared with abdominal, adipose tissue resistance to adrenergic stimulation in vivo as a possible mechanism. Regional fatty acid trafficking, along with the measurement of adipose tissue blood flow (ATBF) with (133)Xe washout, was studied with the arteriovenous difference technique and stable isotope tracers in healthy volunteers. Adrenergic agonists (isoprenaline, adrenaline [epinephrine]) were infused either locally by microinfusion or systemically. Local microinfusion of adrenoceptor antagonists (propranolol, phentolamine) was used to characterise specific adrenoceptor subtype effects in vivo. Femoral adipose tissue NEFA release and ATBF were lower during adrenaline stimulation than in abdominal tissue (p < 0.001). Mechanistically, femoral adipose tissue displayed a dominant α-adrenergic response during adrenaline stimulation. The α-adrenoceptor blocker, phentolamine, resulted in the 'disinhibition' of the femoral ATBF response to adrenaline (p < 0.001). Fatty acids, once stored in femoral adipose tissue, are not readily released upon adrenergic stimulation. Femoral adipose tissue resistance to adrenaline may contribute to the prevention of ectopic fatty acid deposition.

In Vivo Electrochemical Monitoring of the Change of Cochlear Perilymph Ascorbate during Salicylate-Induced Tinnitus

As one of the most important neurochemicals in biological systems, ascorbate plays vital roles in many physiological and pathological processes. In order to understand the roles of ascorbate in the pathological process of tinnitus, this study demonstrates an in vivo method for real time monitoring of the changes of ascorbate level in the cochlear perilymph of guinea pigs during the acute period of tinnitus induced by local microinfusion of salicylate with carbon fiber microelectrodes (CFMEs) modified with multiwalled carbon nanotubes (MWNTs). To accomplish in vivo electrochemical monitoring of ascorbate in the microenvironment of the cochlear perilymph, the MWNT-modified CFME is used as working electrode, a microsized Ag/AgCl is used as reference electrode, and Pt wire is used as counter electrode. Three electrodes are combined together around a capillary to form integrated capillary-electrodes. The integrated capillary-electrode is carefully implanted into the cochlear perilymph of guinea pigs and used both for externally microinfusing of salicylate into the cochlear perilymph and for real time monitoring of the change of ascorbate levels. The in vivo voltammetric method based on the integrated capillary-electrodes possesses a high selectivity and a good linearity for ascorbate determination in the cochlear perilymph of guinea pigs. With such a method, the basal level of cochlear perilymph ascorbate is determined to be 45.0 ± 5.1 μM (n = 6). The microinfusion of 10 mM salicylate (1 μL/min, 5 min) into the cochlear decreases the ascorbate level to 28 ± 10% of the basal level (n = 6) with a statistical significance (P < 0.05), implying that the decrease in ascorbate level in the cochlear may be associated with salicylate-induced tinnitus. This study essentially offers a new method for in vivo monitoring of the cochlear perilymph ascorbate following the salicylate-induced tinnitus and can thus be useful for investigation on chemical essences involved in tinnitus.

Prefrontal and monoaminergic contributions to stop-signal task performance in rats.

Defining the neural and neurochemical substrates of response inhibition is of crucial importance for the study and treatment of pathologies characterized by impulsivity such as attention-deficit/hyperactivity disorder and addiction. The stop-signal task (SST) is one of the most popular paradigms used to study the speed and efficacy of inhibitory processes in humans and other animals. Here we investigated the effect of temporarily inactivating different prefrontal subregions in the rat by means of muscimol microinfusions on SST performance. We found that dorsomedial prefrontal cortical areas are important for inhibiting an already initiated response. We also investigated the possible neural substrates of the selective noradrenaline reuptake inhibitor atomoxetine via its local microinfusion into different subregions of the rat prefrontal cortex. Our results show that both orbitofrontal and dorsal prelimbic cortices mediate the beneficial effects of atomoxetine on SST performance. To assess the neurochemical specificity of these effects, we infused the α2-adrenergic agonist guanfacine and the D(1)/D(2) antagonist α-flupenthixol in dorsal prelimbic cortex to interfere with noradrenergic and dopaminergic neurotransmission, respectively. Guanfacine, which modulates noradrenergic neurotransmission, selectively impaired stopping, whereas blocking dopaminergic receptors by α-flupenthixol infusion prolonged go reaction time only, confirming the important role of noradrenergic neurotransmission in response inhibition. These results show that, similar to humans, distinct networks play important roles during SST performance in the rat and that they are differentially modulated by noradrenergic and dopaminergic neurotransmission. This study advances our understanding of the neuroanatomical and neurochemical determinants of impulsivity, which are relevant for a range of psychiatric disorders.

Attention Deficits and Hyperactivity Following Inhibition of cAMP-Dependent Protein Kinase Within the Medial Prefrontal Cortex of Rats

Previous work demonstrates that microinjections of dopamine D1 receptor agonists and antagonists directly into the medial prefrontal cortex (mPFC) of rats can affect attention in the 5-choice serial reaction time task (5CSRTT), a rodent test analogous to the continuous performance task used to study attention in humans. The present studies were designed to determine if intra-mPFC modulation of cAMP dependent protein kinase (PKA), an intracellular target of D1 receptor stimulation, also affects attention. We examined the effects of localized microinfusions of the cAMP analog Sp-cAMPS (to activate PKA) or Rp-cAMPS (to inhibit PKA) in the 5CSRTT. In parallel we examined the effects of these manipulations on activity levels in an open field, as well as on motivation and the capacity to make complex operant responses using the intracranial self-stimulation (ICSS) test. Inhibition of PKA reduced accuracy in the 5CSRTT and caused substantial increases in locomotor activity without affecting motivation or the capacity to emit operant responses at high rates. Stimulation of PKA also affected some measures of performance in the 5CSRTT, but this effect was associated with reduced capacity to respond at high rates. Viral vector-mediated disruption of cAMP response element binding protein (CREB), a transcription factor directly activated by PKA, also reduced accuracy in the 5CSRTT, raising the possibility that acute inhibition of PKA and sustained inhibition of CREB affect attention through common mechanisms. These studies indicate that PKA inhibition within the mPFC of rats produces inattention and hyperactivity and thus might be useful in modeling human attention disorders.

Intrahippocampal injection of dopamine D1 receptor antagonist SCH23390 interferes with locomotion and learning of active place avoidance, a spatial cognition task

Event Abstract Back to Event Intrahippocampal injection of dopamine D1 receptor antagonist SCH23390 interferes with locomotion and learning of active place avoidance, a spatial cognition task Tomas Petrasek1*, Karel Vales1 and Ales Stuchlik1 1 ASCR, Institute of Physiology, Czechia Neurophysiology of spatial cognition in laboratory rodents is a current topic in behavioral neuroscience. Neurotransmitter substrates are usually studied utilizing the application of specific receptor agonists and antagonists. Contrary to systemic administration, local microinfusions of such drugs have the advantage of selectively targeting the specific brain regions. The present study was aimed at evaluating the role of hippocampal D1 dopamine receptors in performance of Long-Evans rats in an active place avoidance task. This spatial task has been demonstrated to involve cognitive coordination and to be highly dependent upon hippocampal integrity. Adult male rats were implanted with guide cannulae targeted above the dorsal hippocampus. After recovery, the animals were microinfused with a D1 receptor antagonist SCH23390 (at doses 0.5ug, 2ug and 5ug) bilaterally through thin cannula inserts and 30 minutes later tested in the active place avoidance task. Locomotor activity in place avoidance was dose-dependently decreased by SCH23390, whilst the spatial performance (measured by number of entrances into prohibited zone and latency to the first entrance) was found to decrease without a dose-response relationship. These results suggest that dopamine D1 receptors in the hippocampus participate in regulation of locomotion and establishment of a memory trace during initial acquisition in the place avoidance task. This study was supported by GACR grants 309/07/0341, 309/09/0286, IGA MZ CR NR/9178, and MSMT CR 1M0517 and LC554. Conference: 41st European Brain and Behaviour Society Meeting, Rhodes Island, Greece, 13 Sep - 18 Sep, 2009. Presentation Type: Poster Presentation Topic: Poster presentations Citation: Petrasek T, Vales K and Stuchlik A (2009). Intrahippocampal injection of dopamine D1 receptor antagonist SCH23390 interferes with locomotion and learning of active place avoidance, a spatial cognition task. Conference Abstract: 41st European Brain and Behaviour Society Meeting. doi: 10.3389/conf.neuro.08.2009.09.258 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 12 Jun 2009; Published Online: 12 Jun 2009. * Correspondence: Tomas Petrasek, ASCR, Institute of Physiology, Prague, Czechia, disworlds@seznam.cz Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tomas Petrasek Karel Vales Ales Stuchlik Google Tomas Petrasek Karel Vales Ales Stuchlik Google Scholar Tomas Petrasek Karel Vales Ales Stuchlik PubMed Tomas Petrasek Karel Vales Ales Stuchlik Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Time course and efficiency of protein synthesis inhibition following intracerebral and systemic anisomycin treatment

Since its discovery in the 1960s, anisomycin has been used for studying the impact of protein synthesis for manifold cerebral processes such as long-term plastic changes after learning. The common limitation of nearly all pharmacological experiments, including anisomycin treatment, is to precisely verify the affected brain regions. Here we illustrate anisomycin effects on protein synthesis in distinct brain regions of mice (C57BL/6JOlaHsd), revealing differences between three modes of anisomycin application (subcutaneous, s.c.; intraperitoneal, i.p.; local microinfusions into the hippocampus). Our method is based on inhibition of the incorporation of the radioactively-labelled amino acids [ 35S]-Methionine/Cysteine into newly synthesised proteins. Washing the brain slices before autoradiography removes pools of amino acids, which have not been incorporated into newly synthesised proteins, thus, illustrating pure protein synthesis. By comparing different routes of systemic anisomycin application (i.p. versus s.c.; 150 mg/kg) it became evident that the effect of i.p. injection of anisomycin is fully reversed after 6 h, whereas s.c. injection is inhibiting protein synthesis in the hippocampus even 9 h after application. Local microinfusions of anisomycin into the hippocampus were shown to have long-lasting effects as well, which reversed as late as 9 h after injection. The diffusion of anisomycin was maximal at 3 h after injection and more precisely confined to the intended area using a lower dose (20 μg/site) instead of the commonly used dose of 62.5 μg/site. The broad time window of anisomycin action, as revealed in our study, has to be considered, if it comes to the interpretation of time course studies within the context of protein synthesis-dependent processes.