sfn2010

St Peters MM, Bruno JP, Sarter M (2010) Mesolimbic-basal forebrain circuitry mediating the motivational activation of attention. Neuroscience 2010 Abstracts 506.12/LLL52. Society for Neuroscience, San Diego, CA.

Summary: Prefrontal circuitry mediating cue detection is modulated by the tonic component of cholinergic activity. Performance-associated increases in tonic cholinergic activity are augmented by demands on the cognitive control of attention. Highest levels of tonic cholinergic activity are observed while animals perform below baseline as a result of, for example, a distractor, but while they remain motivated to stabilize and recover attentional performance. Cortico-mesolimbic-basal forebrain circuitry is thought to mediate such motivated activation of attentional performance. We previously observed that stimulation of ionotropic glutamate receptors in the shell of the nucleus accumbens (NAs) stimulates tonic cholinergic activity in the prefrontal cortex. Here we test the hypothesis that such stimulation benefits attentional performance while distractors evoke cognitive control. Rats were trained in an operant sustained attention task (SAT) before undergoing surgery for implantation of a bilateral guide cannula targeting the NAs or, in separate animals, the core of the NA (NAc). NMDA (0.01-0.15 µg/0.5 µL/hemisphere) or vehicle (0.9% saline) was infused bilaterally into task-performing animals during SAT and the more challenging distractor version (dSAT). For the dSAT, the operant chamber ceiling lights flashed on/off at 0.5 Hz during the middle block of three blocks of trials that constituted a session. NMDA infusions in the NAs, but not into the NAc, significantly improved the animals’ attentional performance in the presence of the distractor. These findings are consistent with the hypothesis that activation of the NAs mediates attentional performance under conditions that require top-down control. The next set of experiments determined whether the effects of NAs activation require the cortical cholinergic system. We infused the immunotoxin 192 IgG saporin into prefrontal or parietal regions, in addition to implantation of guide cannula targeting the NAs. Replicating the initial finding, NAs NMDA infusions enhanced dSAT performance. Both PFC and PPC cholinergic deafferentation prevented this effect of NMDA. These findings suggest that the motivated activation of the cholinergic attention system during demands on top-down control modulates fronto-parietal attention networks to optimize attentional performance.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Savage ST, Olson L, Mattsson A (2010) Novel object recognition and social interaction in rats lacking cortical cholinergic innervation; comparing manual and digital video tracking systems. Neuroscience 2010 Abstracts 506.9/LLL49. Society for Neuroscience, San Diego, CA.

Summary: Alterations in cholinergic signaling in the brain have been implicated as a contributing factor in the pathogenesis of schizophrenia. We have shown that cholinergic denervation of cortex cerebri by stereotaxic infusion of the immunotoxin 192 IgG-saporin into nucleus basalis magnocellularis in adult rats leads to an enhanced locomotor sensitivity to amphetamine, as well as, a potentiated dopamine release in nucleus accumbens. We have also shown that this cortical cholinergic denervation leads to an increased locomotor response to the NMDA receptor antagonist phencyclidine (PCP), suggesting that disruption of cortical cholinergic activity can lead to disturbances of glutamatergic transmission. We hypothesize that this loss of cortical cholinergic input alters the activity of cortical glutamatergic neurons and in turn, their regulation of subcortical dopamine neurons. In current studies we are investigating memory functions using the novel object recognition task (NOR) and social interaction in adult male Lister hooded rats with cholinergic denervation of neocortex. The behavioral tasks are being conducted under normal conditions and with a PCP-challenge. The data are analyzed both manually by a trained observer, and with a nose point digital video tracking system (Clever Sys Inc.). Manually scoring behavioral data requires extensive observer training, is subject to inter-observer variability, and is time consuming. An automated tracking system could potentially improve upon these issues, however is prone to other problems, including the difficulty of accurately tracking multiple body points. Furthermore, the Lister hooded fur has two different colors which proves difficult for computerized systems to accurately determine the body points. A comparison of the manual scoring and the computerized tracking system is being conducted to determine the most reliable method for each behavioral task. Preliminary results indicate that the cholinergically denervated rats performed the NOR task under normal conditions as well as the controls, however failed to show a preference for the novel object under PCP-challenge. These results were obtained through analysis with both the manual and automated system. Despite fur color difficulties, the video tracking system was able to analyze the NOR task and accurately calculate the distance traveled, which is not easily obtained through manual scoring. These initial results indicate that cortical cholinergic deficits, in addition to a potentiation of the locomotor response to PCP, can also lead to an enhanced sensitivity to PCP-induced cognitive impairments.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Liu Y, Krencik R, Liu H, Ma L, Zhang X, Zhang S-C (2010) Functional cholinergic neurons from human embryonic stem cells. Neuroscience 2010 Abstracts 331.5/B19. Society for Neuroscience, San Diego, CA.

Summary: Basal forebrain cholinergic neurons play a critical role in regulating memory and cognition. Degeneration or dysfunction of these neurons is associated with neurological conditions including Alzheimer’s disease and dementia. In this study, we aimed at generating cholinergic neurons from human embryonic stem cells (hESCs) for therapeutic development. hESCs were first differentiated to primitive neuroepithelial cells in a chemically defined medium. In the presence of sonic hedgehog, over 97% of the differentiated cells became Nkx2.1-expressing ventral forebrain progenitors. These ventral progenitors further differentiated to cholinergic neurons with basal forebrain characteristics by expressing ChAT, VAChAT, FoxG1, Nkx2.1, Islet1, ßIII-tubulin, MAP2, P75, Synapsin but not GABA, Glutamate, or Mash2. The hESC-generated cholinergic neurons were electrophysiologically active in vitro. Following transplantation into the hippocampus of mice, in which cholinergic neurons in the medial septum were destroyed by IgG-P75-saporin, the grafted human cells produced large cholinergic neurons. The animals transplanted with cholinergic neurons demonstrated an improvement in learning and memory deficit. These results indicate that the human stem cell-generated cholinergic neurons are functional, thus providing a new source for drug discovery and cell therapy for neurological disorders that affect cholinergic neurons.

Related Products: mu p75-SAP (Cat. #IT-16)

ATS Poster of the Year Winner

Rennie KE, Frechette M, Pappas BA (2010) The effects of neonatal cholinergic lesion on age-related changes in behaviour, neurogenesis and CA1 pyramidal cell morphology. Neuroscience 2010 Abstracts 349.8/J12. Society for Neuroscience, San Diego, CA.

Summary: Age-related cognitive decline is associated with dysfunction of the basal forebrain cholinergic (BFC) system, and cortico-hippocampal cholinergic denervation is a hallmark neurochemical feature of the Alzheimer’s-afflicted brain. It has been suggested that cognitive deficits that emerge with age may be rooted in early dysfunction of the BFC system and that impaired cholinergic transmission might interact with ageing-associated factors to produce cognitive decline. The purpose of this study was to examine the effects of neonatal cholinergic lesion on age-related changes in spatial working memory, neurogenesis and hippocampal CA1 pyramidal cell morphology. We have previously reported that neonatal cholinergic lesion results in only minor behavioural deficits, but impairs the birth and/or survival of new neurons and reduces CA1 dendritic complexity in the young adult rat. We hypothesized that memory impairments would become apparent in lesioned rats as they age, and that this impairment would be accompanied by more drastic reductions in neurogenesis and cytoarchitectural alterations than those that have been documented in the young adult animal after neonatal cholinergic lesion. Seven-day-old male Sprague-Dawley rats were subjected to basal forebrain cholinergic lesion by infusion of the cholinotoxin 192-IgG-Saporin into the lateral ventricles. At the age of 12 or 21 months, the rats were tested on a working memory version of the Morris water maze. While aging had only a slight effect on the memory performance of control rats, lesioned rats showed pronounced memory impairments with age. This occurred without CA1 cell loss or astrogliosis in 21-month-old lesioned rats when compared to age-matched controls. However, golgi analysis revealed that while cholinergic lesion did not alter the total dendritic length, branching, number of spines, or spine density of CA1 pyramidal cells in 21-month-old rats, the distribution of these parameters across branch orders was shifted. The lesion caused a slight reduction in apical branch length and spine density, and basal branch number, length and number of spines at low/middle branch orders, but increased these parameters at upper branch orders. Thus, perinatal cholinergic lesion precipitates spatial memory dysfunction during old age, and this seems to be associated with cytoarchitectural changes to neurons rather than neuronal loss.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Li A-J, Wang Q, Dinh TT, Ritter S (2010) Hindbrain catecholamine neurons are required for rapid switching of metabolic substrate utilization during glucoprivation. Neuroscience 2010 Abstracts 392.14/III1. Society for Neuroscience, San Diego, CA.

Summary: Glucoprivation is a metabolic emergency in which a rapid and effective system-wide switch to fat metabolism must occur to conserve any available glucose for use by the brain. Glucoprivation stimulates secretion of corticosterone, which is known to play an important role in promoting fat utilization. In previously published work, we showed that injections of the retrogradely transported catecholamine neuron immunotoxin, anti-dopamine beta-hydroxylase conjugated to saporin (DSAP) into the paraventricular nucleus of the hypothalamus (PVH) eliminate the corticosterone response to glucoprivation without impairing the response to a nonmetabolic stressor (swim stress), without altering the circadian rhythm of corticosterone secretion and without damaging the PVH CRF-secreting neurons. Here we microinjected DSAP into the PVH to selectively lesion hindbrain catecholamine neurons innervating this site, thus impairing the glucoprivation-induced corticosterone response. Using indirect calorimetry, we examined metabolic fuel utilization and other metabolic parameters in these lesioned rats under basal and glucoprivic conditions. Under basal conditions, energy expenditure and locomotor activity did not differ between DSAP rats and controls injected with unconjugated saporin (SAP). However, DSAP rats had a higher respiratory exchange ratio (RER) than SAPs, indicating their greater dependence on carbohydrate utilization. Glucoprivation induced by 2-deoxy-D-glucose (2DG, 250 mg/kg) reduced energy expenditure equally in SAP and DSAP rats. However, 2DG rapidly decreased RER to 0.8 (a value indicating ongoing fat metabolism) in the SAP group, but not in the DSAP group. Responses to 2DG persisted for about 6 hours. Adrenal dennervation, which eliminates the adrenal medullary response to glucoprivation, did not alter these responses to 2DG in either SAP or DSAP rats. Results indicate that in the absence of hypothalamically-projecting hindbrain catecholamine neurons, rats cannot efficiently switch their fuel utilization from carbohydrate to fat during glucoprivation, presumably due to a deficient corticosterone response. Results also suggest a previously unrecognized role for these catecholamine neurons in control of basal substrate utilization.

Related Products: Anti-DBH-SAP (Cat. #IT-03)

Jeong D, Lee D, Chang J (2010) Decrease of Arc protein expression and delay of memory acquisition by immunolesion. Neuroscience 2010 Abstracts 145.5/H6. Society for Neuroscience, San Diego, CA.

Summary: Cholinergic neuronal deficit is one of the common characteristics in both Alzheimer’s disease dementia (AD) and vascular dementia (VaD). Forebrain Cholinergic neurons in the basal forebrain project to the neocortex and the hippocampus which make an important role in memory function. We used 192 IgG-saporin to produce selective lesion of cholinergic basal forebrain neurons including the medial septum (MS) and the nucleus basalis magnocellularis (NBM). We intracerebroventricularly injected 192 IgG-saporin (0.63 µg/µl dose, 6 µl, 8 µl and 10 µl) or phosphate buffered saline (8 µl). Morris water maze and tissue perforation for immunohistochemistry and western blotting were sequentially performed 2 weeks after injection of 192 IgG-saporin. In the acquisition phase of Morris water maze, latency of 6ul group (2nd day), 8 µl group (2nd day) and 10 µl group (3rd day) was significantly delayed but it was recovered within 1week. Time in platform and the number of crossing were significantly different between 8 µl LV injection group and sham group in probe test. In immunohistological study, the extent of the cholinergic lesion was showed in the basal forebrain complex region of all 192 IgG-saporin injected rats. Expression of Arc protein is significantly decreased in the frontal cortex (8 µl and 10 µl groups) but hippocampus. Decrease of parvalbumin in the frontal cortex (8ul and 10 ul groups) and the hippocampus (10 µl) means nonselective lesion because of high dose of immunotoxin. We observed recovery after memory acquisition delay and decrease of synaptic activity in the frontal cortex except in the hippocampus. High dose of immunotoxin injured not only cholinergic neuron but also GABAergic neuron in the frontal cortex and the hippocampus. Hippocampal GABAergic cell synapse on to glutamatergic pyramidal cells. Deficit of the hippocampal inhibitory cell may facilitate hippocampal synaptic plasticity and the recovery.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Szot P, Franklin A, White S, Raskind M (2010) Time- and dose-response of 6-hydroxydopamine on locus coeruleus noradrenegric neurons in c57bl/6 mice. Neuroscience 2010 Abstracts 157.20/R1. Society for Neuroscience, San Diego, CA.

Summary: Locus coeruleus (LC) noradrenergic neurons are severely reduced in Alzheimer’s and Parkinson’s disease. However, it is unclear why these neurons are lost and the consequence of this loss on the progression and symptoms of these neurodegenerative disorders. Therefore, establishing an animal model of LC noradrenergic neuronal loss is critical in determining how the LC contributes to these disorders. The purpose of this study was to determine the dose- and time-response of noradrenergic neurotoxicity of 6-hydroydopamine (6OHDA) in adult male C57BL/6 mice. Our laboratory recently demonstrated that DSP4 does not result in a loss of LC noradrenergic neurons. Neurotoxicity of 6OHDA on LC noradrenergic neurons was determined by measuring tyrosine hydroxylase (TH) mRNA expression and TH-immunoreactivity (IR) in LC noradrenergic neurons. TH mRNA was quantitated using MCID (OD), while TH-IR was used to determine if protein levels reflected what was observed with mRNA. 6OHDA (20 µg/µl bilaterally) and dopamine beta-hydroxylase-saporin (DBH-saporin; 1 µg/µl bilaterally) were initially administered into the lateral ventricles (icv) and sacrificed 2 weeks later. 6OHDA reduced TH mRNA and -IR in both the dopaminergic neurons of the substantia nigra (SNpc) and ventral tegmental nucleus (VTA), and LC by -46%, -65% and -63%, respectively. DBH-saporin icv injection did not affect dopaminergic or noradrenergic neurons. Injection of DBH-saporin into the LC (0.1 µg/µl unilaterally) also did not affect LC noradrenergic neurons 2 weeks later. As a time-course 6OHDA (7 µg/µl) was injected unilaterally into the LC (vehicle was administered in the alternate LC) and sacrificed 3 days, 2 and 3 weeks later. A loss of LC noradrenergic neurons was observed only 3 weeks later (-81.4%). 6OHDA was then injected unilaterally into the LC at 7, 10, and 14 ug/ul (vehicle was administered in the alternate LC) and sacrificed 2 weeks later. The 7 µg/µl dose of 6OHDA did not affect TH mRNA in the LC as compared to control side (-19%), 10 ug/ul 6OHDA significantly reduced TH mRNA in the LC by ~55%, and 14 ug/ul 6OHDA dramatically reduced TH mRNA in the LC by ~90%. TH-IR in the LC of the three different 6OHDA doses reflected closely the TH mRNA data. 6OHDA at the dose of 14 µg/µl, which resulted in a near complete loss of LC noradrenergic neurons, did not affect dopaminergic neurons in the SN (-9%) and VTA (+17%). These data indicate that DBH-saporin, at the parameters studied, did not affect mouse LC noradrenergic neurons. 6OHDA demonstrated a time- and dose-response reduction of mouse LC noradrenergic neurons. The consequence of this LC neuronal loss on forebrain noradrenergic markers will also be presented.

Related Products: Anti-DBH-SAP (Cat. #IT-03)

Datta S, Chatterjee K, Wiley R (2010) Decreasing abnormal nocifensive responses in the bilateral chronic constriction injury (bCCI) model of neuropathic pain: Effects of lumbar intrathecal CCK-saporin. Neuroscience 2010 Abstracts 175.22/MM12. Society for Neuroscience, San Diego, CA.

Summary: The bCCI model produces long lasting -cold hyperalgesia (at least 100 days) along with decreases in staining for cholecystokinin (CCK) in the dorsal horn (DH). Spinal cholecystokinin (CCK) has anti-opiate activity, and selective destruction of DH neurons expressing CCK receptors by injection of intrathecal CCK-saporin, in naïve rats decreases thermal nocifensive reflex responses and is additive with morphine in decreasing nocifensive responses to heat. In the present study, we sought to determine the effects of intrathecal CCK-sap in the bCCI model of neuropathic pain in Long Evans female rats. bCCI rats underwent bilateral ligation of the sciatic nerves with chromic gut sutures. Controls underwent sham surgery with no ligation. Rats were tested on 0.3 C cold plate, thermal preference task (TPT) (shuttle box with floor temperatures of 15 C vs 45 C) and mechanical stimulation (von Frey). bCCI produced increased responses on the cold plate. 21 days after the bCCI surgery, the rats were injected with 1500 ng CCK-sap into the lumber CSF. Then, thermal and mechanical testing was repeated at intervals. Intrathecal CCK-sap injections decreased abnormal nocifensive responding of bCCI rats on the cold plate. CCK-sap reduced withdrawal responses to mechanical stimulation in bCCI rats. In TPT testing, the bCCI animals were hyperalgesic to cold (reduced cold side occupancy). After intrathecal CCK-sap injections, thermal preference was reversed (increased cold side occupancy). We interpret these results as showing that CCK-sap reverses abnormal nocifensive responses of bCCI in rats to aversive cold and mechanical stimuli. These results suggest that silencing CCK receptor-expressing superficial DH neurons is a potential strategy for development of new treatments for chronic neuropathic pain.

Related Products: CCK-SAP (Cat. #IT-31)

Li C, Back S, Lee J, Baek SK, Na H (2010) Gastrin-releasing peptide receptor in the spinal cord mediates mechanical allodynia following nerve injury. Neuroscience 2010 Abstracts 176.2/OO4. Society for Neuroscience, San Diego, CA.

Summary: Gastrin-releasing peptide receptor (GRPR) has been suggested as an itch-specific gene in the spinal cord (Sun et al., Nature, 2009). They described that selective ablation of GRPR-expressing lamina I neurons led to deficits in itch-related scratching behaviors without any effects on pain behaviors including nerve injury-induced mechanical allodynia. It has been known that two types of mechanical allodynia, such as static and dynamic allodynia, can be detectable in neuropathic patients, and may be mediated by distinct mechanisms. In the present study, we investigated the role of spinal GRPR in each of static and dynamic allodynia using both rat- and mouse-tail models of neuropathic pain. Bombesin-saporin (bombesin-sap) was administered intrathecally to ablate spinal GRPR-expressing neurons. Scratching behaviors evoked by pruritogenic agents, such as serotonin and chloroquine, and physiological pain behaviors were analyzed before nerve injury. Static or dynamic allodynia was assessed by the application of von Frey filaments to the tail or brushing the tail with a filament, respectively. RC3095, a GRPR antagonist, was given intrathecally to see its effects on static and dynamic allodynia in neuropathic rats. Bombesin-sap treatment resulted in reduction of GRPR-immunoreactive cells in lamina I of spinal dorsal horn and scratching deficits. Physiological pain behaviors of these animals were not different from those of control animals. Following the partial injury of tail-innervating nerves, animals treated with bombesin-sap exhibited comparable dynamic allodynia to control one. However, they failed to manifest static allodynia during the entire experimental period. In addition, RC3095 relieved static, but not dynamic, allodynia. These findings suggest that spinal GRPR mediates nerve injury-induced static mechanical allodynia as well as itching sensation in normal state.

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Gulino R, Gulisano M (2010) The recovery of locomotion after lumbar spinal cord motoneuron depletion is affected by the modulation of Sonic Hedgehog and Notch-1 pathways. Neuroscience 2010 Abstracts 259.19/W12. Society for Neuroscience, San Diego, CA.

Summary: Sonic hedgehog (Shh) and Notch-1 are involved in the regulation of stem cell function. Additionally, Notch-1 has a role as modulator of synaptic plasticity. In our previous work, we injected Cholera toxin-B saporin (CTB-sap) into the gastrocnemius muscle to induce a selective depletion of motoneurons within lumbar mice spinal cord (SC) and analysed the expression levels of Shh, Notch-1, Numb, Choline acetyltransferase (ChAT) and Synapsin-I proteins. The functional outcome of the lesion was monitored by grid walk test and rotarod. Shh and Notch-1 appeared reduced in the lesioned tissue and correlated with ChAT and Synapsin-I levels, suggesting a role in modulating synaptic plasticity. Numb expression was also reduced after lesion and appeared correlated with motor performance but not with synaptic plasticity. We sought to determine if the pharmacological manipulation of the expression of Shh and Notch-1 could affect functional recovery by a mechanism involving synaptic plasticity. Therefore, we used CTB-sap to induce lesion as above, and injected Cyclopamine, recombinant Shh or recombinant DLL4 chronically into the intrathecal space by means of osmotic minipumps for two weeks. The functional recovery was monitored for one month after lesion by means of grid walk test, two times a week, whereas the modifications of Shh, Notch-1, ChAT and Synapsin-I protein expression levels were measured by western blot and immunohistochemistry. Here, we show that the modulation of Shh or Notch-1 pathways could affect the recovery of locomotion. Moreover, the molecular mechanisms underlying this process is discussed.

Related Products: CTB-SAP (Cat. #IT-14)