Shaddock (Citrus maxima) peels extract restores cognitive function, cholinergic and purinergic enzyme systems in scopolamine-induced amnesic rats.

This study investigated the effect of shaddock peels extract on cognitive function in scopolamine-induced amnesic rats. Wistar rats were pretreated with shaddock peels extract (50 and 100 mg/kg) and donepezil (5 mg/kg) for fourteen days via oral administration. Memory impairment was induced at the end of the treatment period via a single intraperitoneal administration of scopolamine (3 mg/kg). Thereafter, the animals were subjected to behavioral studies (Morris water maze and Y-maze tests). Finally, the rats were sacrificed and the hippocampus of the rat's brain was isolated for biochemical analyses. The results showed a significant decrease in memory and cognitive function as revealed by Morris water maze and Y-maze tests in scopolamine-induced rats which were reversed by shaddock peels extract. Also, there was a significant decrease in the activity of adenosine monophosphohydrolase (AMPase) with a simultaneous increase in activities of adenosine deaminase (ADA), adenosine triphosphate diphosphohydrolase (ATPdase), acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in scopolamine-induced rats when compared with the control. Besides, a significant increase in malondialdehyde (MDA) and reactive oxygen species (ROS) levels were observed in scopolamine-induced rats. However, donepezil or shaddock peels extract (50 and 100 mg/kg) caused a significant inhibitory effect on AChE, and ADA activities when compared to scopolamine-induced rats. Rats treated with shaddock peels extract also showed a significant reduction in MDA and ROS levels compared to scopolamine-induced rats. Therefore, our findings showed that the cognitive-enhancing effects of shaddock peels extract could be due to antioxidant activities and modulation of some enzymes linked with cognitive dysfunction.

Sublethal doses of neonicotinoid imidacloprid can interact with honey bee chemosensory protein 1 (CSP1) and inhibit its function.

As a frequently used neonicotinoid insecticide, imidacloprid can impair the chemoreceptive behavior of honey bees even at sublethal doses, while the physiochemical mechanism has not been further revealed. Here, multiple fluorescence spectra, thermodynamic method, and molecular docking were used to study the interaction and the functional inhibition of imidacloprid to the recombinant CSP1 protein in Asian honey bee, Apis cerana. The results showed that the fluorescence intensity (λem = 332 nm) of CSP1 could be significantly quenched by imidacloprid in a dynamic mode. During the quenching process, ΔH > 0, ΔS > 0, indicating that the acting forces of imidacloprid with CSP1 are mainly hydrophobic interactions. Synchronous fluorescence showed that the fluorescence of CSP1 was mainly derived from tryptophan, and the hydrophobicity of tryptophan decreased with the increase of imidacloprid concentration. Molecular docking predicted the optimal pose and the amino acid composition of the binding process. Circular dichroism (CD) spectra showed that imidacloprid reduced the α-helix of CSP1 and caused the extension of the CSP1 peptide chain. In addition, the binding of CSP1 to floral scent ß-ionone was inhibited by nearly 50% of the apparent association constant (KA) in the presence of 0.28-2.53 ng/bee of imidacloprid, and the inhibition rate of nearly 95% at 3.75 ng/bee of imidacloprid at sublethal dose level. This study initially revealed the molecular physiochemical mechanism that sublethal doses of neonicotinoid still interact and inhibit the physiological function of the honey bees' chemoreceptive system.

Midazolam-ketamine dual therapy stops cholinergic status epilepticus and reduces Morris water maze deficits.

OBJECTIVE: Pharmacoresistance remains an unsolved therapeutic challenge in status epilepticus (SE) and in cholinergic SE induced by nerve agent intoxication. SE triggers a rapid internalization of synaptic γ-aminobutyric acid A (GABAA ) receptors and externalization of N-methyl-d-aspartate (NMDA) receptors that may explain the loss of potency of standard antiepileptic drugs (AEDs). We hypothesized that a drug combination aimed at correcting the consequences of receptor trafficking would reduce SE severity and its long-term consequences. METHODS: A severe model of SE was induced in adult Sprague-Dawley rats with a high dose of lithium and pilocarpine. The GABAA receptor agonist midazolam, the NMDA receptor antagonist ketamine, and/or the AED valproate were injected 40 min after SE onset in combination or as monotherapy. Measures of SE severity were the primary outcome. Secondary outcomes were acute neuronal injury, spontaneous recurrent seizures (SRS), and Morris water maze (MWM) deficits. RESULTS: Midazolam-ketamine dual therapy was more efficient than double-dose midazolam or ketamine monotherapy or than valproate-midazolam or valproate-ketamine dual therapy in reducing several parameters of SE severity, suggesting a synergistic mechanism. In addition, midazolam-ketamine dual therapy reduced SE-induced acute neuronal injury, epileptogenesis, and MWM deficits. SIGNIFICANCE: This study showed that a treatment aimed at correcting maladaptive GABAA receptor and NMDA receptor trafficking can stop SE and reduce its long-term consequences. Early midazolam-ketamine dual therapy may be superior to monotherapy in the treatment of benzodiazepine-refractory SE.

Histamine release in the basal forebrain mediates cortical activation through cholinergic neurons.

The basal forebrain (BF) is a key structure in regulating both cortical activity and sleep homeostasis. It receives input from all ascending arousal systems and is particularly highly innervated by histaminergic neurons. Previous studies clearly point to a role for histamine as a wake-promoting substance in the BF. We used in vivo microdialysis and pharmacological treatments in rats to study which electroencephalogram (EEG) spectral properties are associated with histamine-induced wakefulness and whether this wakefulness is followed by increased sleep and increased EEG delta power during sleep. We also investigated which BF neurons mediate histamine-induced cortical activation. Extracellular BF histamine levels rose immediately and remained constant throughout a 6 h period of sleep deprivation, returning to baseline levels immediately afterward. During the spontaneous sleep-wake cycle, we observed a strong correlation between wakefulness and extracellular histamine concentrations in the BF, which was unaffected by the time of day. The perfusion of histamine into the BF increased wakefulness and cortical activity without inducing recovery sleep. The perfusion of a histamine receptor 1 antagonist into the BF decreased both wakefulness and cortical activity. Lesioning the BF cholinergic neurons abolished these effects. Together, these results show that activation of the cholinergic BF by histamine is important in sustaining a high level of cortical activation, and that a lack of activation of the cholinergic BF by histamine may be important in initiating and maintaining nonrapid eye movement sleep. The level of histamine release is tightly connected to behavioral state, but conveys no information about sleep pressure.

Neurotranmission systems as targets for toxicants: a review.

Neurotransmitters are chemicals that transmit impulses from one nerve to another or from nerves to effector organs. Numerous neurotransmitters have been described in mammals, amongst them acetylcholine, amino acids, amines, peptides and gases. Toxicants may interact with various parts of neurotransmission systems, including synthetic and degradative enzymes, presynaptic vesicles and the specialized receptors that characterize neurotransmission systems. Important toxicants acting on the cholinergic system include the anticholinesterases (organophosphates and carbamates) and substances that act on receptors such as nicotine and the neonicotinoid insecticides, including imidacloprid. An important substance acting on the glutamatergic system is domoic acid, responsible for amnesic shellfish poisoning. 4-Aminobutyric acid (GABA) and glycine are inhibitory neurotransmitters and their antagonists, fipronil (an insecticide) and strychnine respectively, are excitatory. Abnormalities of dopamine neurotransmission occur in Parkinson's disease, and a number of substances that interfere with this system produce Parkinsonian symptoms and clinical signs, including notably 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, which is the precursor of 1-methyl-4-phenylpyridinium. Fewer substances are known that interfere with adrenergic, histaminergic or seroninergic neurotransmission, but there are some examples. Among peptide neurotransmission systems, agonists of opioids are the only well-known toxic compounds.

Tramadol safety--cholinergic system of rat brain without nociception.

In the present investigation, changes in the levels of acetylcholinesterase (AChE) activity, acetylcholine (ACh) content, and the activity levels of plasma (PChE) and erythrocyte (EChE) cholinesterases as representatives of pseudocholinesterases were examined in different areas of the rat brain during the administration of the synthetic opioid analgesic drug tramadol (Ultram) without induction of pain. Male adult Wistar rats weighing 150 +/- 20 g were used. Tramadol was injected subcutaneously (s.c.) into the rats at 0, 24 and 48 h, and the changes in the above cholinergic parameters were recorded after the completion of 3, 6, 12, 24, 48 and 72 h. Following administration of single dose (for rats sacrificed at 24 h) and multiple doses (for rats sacrificed at 48 and 72 h) of tramadol, the ACh content showed an increase in all brain areas. Concurrently, the AChE activity was found to decrease in all the areas. PChE and EChE showed higher activity levels, with EChE showing a higher level of activity than PChE. The levels of all the parameters examined returned towards the control levels by about 24 h after the administration of single dose of tramadol. However, the ACh levels showed an elevation at 48 and 72 h (following double and triple doses, respectively). The AChE activity levels also showed a simultaneous increase at 48 and 72 h, presumably to balance the increase in ACh levels on longer treatment with tramadol. The observed changes in the cholinergic segment presumably do not cause any physiological lesion since they reverted to control levels after the time limit of change under tramadol influence. This observation indicates that tramadol can be administered safely both under nociceptive and non-nociceptive conditions.

Integrated transcriptomic and proteomic analysis indicated that neurotoxicity of rats with chronic fluorosis may be in mechanism involved in the changed cholinergic pathway and oxidative stress.

BACKGROUND: To reveal the underling molecular mechanism in brain damage induced by chronic fluorosis, the neurotoxicity and its correlation were investigated by transcriptomics and proteomics. METHODS: Sprague-Dawley rats were treated with fluoride at different concentrations (0, 5, 50 and 100 ppm, prepared by NaF) for 3 months. Spatial learning and memory were evaluated by Morris water maze test; neuronal morphological change in the hippocampus was observed using Nissl staining; and the level of oxidative stress including reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were detected by biological methods. The high-throughput transcriptome sequencing (RNA-Seq) and tandem mass tag (TMT) proteomic sequencing were performed to detect the expression of differentially expressed genes and proteins, respectively. RESULTS: The results showed that compared with control group, rats exposed to high-dose fluoride exhibited declined abilities of learning and memory, decreased SOD activity and increased ROS and MDA levels, with lighter colored Nissl bodies. A total of 28 important differentially expressed genes (DEGs) were screened out by transcriptomics. Then, functional enrichment analyses showed that upregulated proteins enriched in cellular transport, while downregulated proteins enriched in synapse-related pathways. Thirteen corresponding DEGs and DAPs (cor-DEGs-DAPs) were identified by differential expressions selected with positively correlated genes/proteins, most of which were related to neurodegenerative changes and oxidative stress response. CONCLUSION: These results provide new omics evidence that rats chronically exposed to high-dose fluoride can induce neurotoxicity in the brains through changes in the cholinergic pathway and oxidative stress.

Cholinergic deafferentation of the neocortex using 192 IgG-saporin impairs feature binding in rats.

The binding problem refers to the fundamental challenge of the CNS to integrate sensory information registered by distinct brain regions to form a unified neural representation of a stimulus. Although the human cognitive literature has established that attentional processes in frontoparietal cortices support feature binding, the neurochemical and specific downstream neuroanatomical contributions to feature binding remain unknown. Using systemic pharmacology in rats, it has been shown that the neuromodulator acetylcholine is essential for feature binding at encoding, but the neural source of such critical cholinergic neurotransmission has yet to be identified. Cholinergic efferents from the nucleus basalis magnocellularis (NBM) of the basal forebrain provide the majority of the cholinergic input to the neocortex. Accordingly, it was hypothesized that the NBM is the neural source that provides the critical neuromodulatory support for feature binding. To test this hypothesis, rats received bilateral 192 IgG-saporin lesions of the NBM, and their feature binding performance was tested using a forced-choice digging paradigm. Relative to sham-lesioned rats, NBM-lesioned rats were significantly impaired at acquiring a crossmodal feature conjunction (FC) stimulus set that required feature binding, whereas their ability to retrieve an FC stimulus set and to acquire two crossmodal feature singleton stimulus sets, one of greater difficulty than the other but neither requiring feature binding, remained intact. These behavioral findings, along with histological analyses demonstrating positive relationships between feature-binding acquisition and markers of cholinergic activity in frontoparietal regions, reveal the importance of neocortical cholinergic input from the NBM to feature binding at encoding.

Caenorhabditis elegans bicarbonate transporter ABTS-1 is involved in arsenite toxicity and cholinergic signaling.

Arsenic poisoning affects millions of people worldwide. Although there is accumulating evidence to suggest that the nervous system is a target of arsenic, relatively little information is known regarding its effects on the nervous system. The effects of arsenite on the nervous system in Caenorhabditis elegans were investigated in the present study. We found that abts-1, which encodes a Na(+)-dependent Cl(-)/HCO(3)(-) transporter, is required to protect C. elegans from arsenite toxicity. The abts-1::GFP transgene is primarily expressed in neurons and the hypodermis, but stronger expression was also observed in the pharynx and body wall muscle cells after exposure to arsenite. The steady-state level of ABTS-1 mRNA increased in response to arsenite exposure. We showed that worms lacking abts-1 are hypersensitive to the paralytic effects of the cholinesterase inhibitor, aldicarb, and the nicotinic acetylcholine receptor agonist, levamisole. We also showed that arsenite enhanced sensitivity to aldicarb and levamisole in abts-1 mutant worms. Our results indicate neuronal effects of arsenite and the ABTS-1 bicarbonate transporter.

Crystal structure of a catalytically active, non-toxic endopeptidase derivative of Clostridium botulinum toxin A.

Botulinum neurotoxins (BoNTs) modulate cholinergic nerve terminals to result in neurotransmitter blockade. BoNTs consists of catalytic (LC), translocation (Hn) and cell-binding domains (Hc). The binding function of the Hc domain is essential for BoNTs to bind the neuronal cell membrane, therefore, removal of the Hc domain results in a product that retains the endopeptidase activity of the LC but is non-toxic. Thus, a molecule consisting of LC and Hn domains of BoNTs, termed LHn, is a suitable molecule for engineering novel therapeutics. The structure of LHA at 2.6 A reported here provides an understanding of the structural implications and challenges of engineering therapeutic molecules that combine functional properties of LHn of BoNTs with specific ligand partners to target different cell types.

Spatial memory following selective cholinergic lesion of the nucleus basalis magnocellularis.

The aim of this study was to investigate the modulation of the cognitive function by the cholinergic cells of the nucleus basalis magnocellularis (NBM) and was designed to investigate the role of the NBM cholinergic cells in learning and memory using the immunotoxin 192 IgG-saporin to produce selective lesions of cholinergic NBM neurons. A total of 16 male outbred albino rats were used in the present study to investigate the ability of sham-operated and NBM immunotoxin lesioned rats to learn the location of a visible, as well as submerged platform in a water maze. Examination of the AChE stained sections showed that after injections of 192 IgG saporin into the NBM, animals exhibited significantly less AChE staining in PFC as compared to sections obtained from sham-operated animals. An overview of the data from both competition trials for each group show that the sham-operated rats in 13 trials out of 16 competition test trial used place strategy and NBM-lesioned ones used this strategy in 6 trials. Decreased place-bias in NBM-lesioned rats compared to the sham-operated rats was significant (t(d )= 2,42, P<0.02).The data obtained in the sham-operated and NBM-lesioned animals in the present study, demonstrate that the choice of strategy in the competition trial is related to performance during training: the rats exhibiting cue strategy (NBM-lesioned) on the competition trial had significantly worse performance during hidden platform training than those (sham-operated) exhibiting a place strategy. These findings suggest that the NBM is essential for accurate spatial learning and suggest its role in processing information about the spatial environment, but also we can propose, that the behavioral deficits described in the present study is nonmnemonic, possibly caused by deficit in attentional function.

Reduced cholinergic status in hippocampus produces spatial memory deficits when combined with kainic acid induced seizures.

Alzheimer's disease is the most common form of dementia in North America today. Though many risk factors have been suggested to increase the likelihood of developing this disease, an accurate etiology has yet to be described. One of these risk factors commonly associated with Alzheimer's disease is the loss of cholinergic neurons of the medial septum that project to the hippocampus, leading to depletion in cholinergic activity. A second risk factor is the presence of seizures, which can increase the risk of excitotoxic cell death. To examine the interaction between these two common risk factors, we gave rats a focal cholinergic lesion of the medial septum using the specific immunotoxin 192-IgG Saporin, followed 2 weeks later by a non-convulsive dose of kainic acid. We then assessed the rats for seizure severity, hippocampal damage and performance on a spatial memory task. The combination of the two factors resulted in a trend towards increased seizure severity in the cholinergic depleted rats, but more importantly, the lesioned rats that had non-convulsive seizures were significantly impaired on a spatial version of the Morris water maze when compared with either the rats with a cholinergic depletion or non-convulsive seizure alone. This result could not be explained by seizure severity or the extent of hippocampal damage, suggesting a more subtle interaction between these two risk factors in the development of a hippocampal based memory impairment.

Emergence of spatial impairment in rats following specific cholinergic depletion of the medial septum combined with chronic stress.

A consistent finding in patients suffering from Alzheimer's disease is a loss of the cholinergic neurons of the basal forebrain that project to the hippocampus. However, the role this depletion plays in the development of Alzheimer's disease remains unclear. The loss of this ascending neurotransmitter system could potentially render hippocampal neurons more susceptible to further insult, such as chronic stress, ultimately resulting in neuronal death and memory loss. We explored this possibility by using the highly specific toxin 192 IgG-Saporin to destroy the majority of cholinergic activity in the septo-hippocampal pathway in rats. Following depletion, rats were subjected to 2 weeks of restraint stress. Rats were divided into two groups and were tested either on a hippocampal-dependent (water maze) task or a hippocampal-independent task (fear conditioning to tone and context). We showed that cholinergic depletion or stress alone had no effect on the successful performance of either of the tasks. However, rats with a combination of cholinergic depletion and stress were significantly impaired on the water-maze task. No deficits were apparent in the combined group that was tested on fear conditioning to tone or context, suggesting that this impairment is specific to spatial working memory. These rats had no obvious hippocampal neuronal loss or damage; however, there were likely subtle changes in hippocampal processing that led to the observed deficit on the hippocampal-dependent task. These findings support our theory that cholinergic depletion of the medial septum increases hippocampal vulnerability to further insults such as stress.

Acrylamide toxicity and cholinergic nervous system.

Acrylamide (ACR) is a chemical compound, that forms in starchy food products during cooking at high-temperatures, including frying, baking, and roasting. ACR is a known lethal neurotoxin. The presented review suggests that the mechanism of ACR's neurotoxicity may be related to an impaired cholinergic transmission in the central and peripheral nervous system and redox imbalance. These may not only affect ongoing brain functions but also participate in etiology of neurodegeneration.

Toxicokinetics of the neonicotinoid insecticide imidacloprid in rainbow trout (Oncorhynchus mykiss).

Studies were conducted to determine the distribution and elimination of imidacloprid (IMI) in rainbow trout. Animals were injected with a low (47.6 µg/kg), medium (117.5 µg/kg) or high (232.7 µg/kg) dose directly into the bloodstream and allowed to depurate. The fish were then sampled to characterize the loss of IMI from plasma and its appearance in expired water (all dose groups) and urine (medium dose only). In vitro biotransformation of IMI was evaluated using trout liver S9 fractions. Mean total clearance (CLT) values determined by non-compartmental analysis of plasma time-course data were 21.8, 27.0 and 19.5 mL/h/kg for the low, medium and high dose groups, respectively. Estimated half-lives for the same groups were 67.0, 68.4 and 68.1 h, while fitted values for the steady-state volume of distribution (VSS) were 1.72, 2.23 and 1.81 L/kg. Branchial elimination rates were much lower than expected, suggesting that IMI is highly bound in blood. Renal clearance rates were greater than measured rates of branchial clearance (60% of CLT in the medium dose group), possibly indicating a role for renal membrane transporters. There was no evidence for hepatic biotransformation of IMI. Collectively, these findings suggest that IMI would accumulate in trout in continuous waterborne exposures.

Effect of N-methyl-d-aspartate receptor blockade on plasticity of frontal cortex after cholinergic deafferentation in rat.

Cholinergic projections from the nucleus basalis play a critical role in cortical plasticity. For instance, cholinergic deafferentation increases dendritic spine density and expression of the GluR1 subunit of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate receptor in frontal cortex. Acetylcholine modulates glutamatergic activity in cortex, and the N-methyl-d-aspartate subtype of glutamate receptor plays a role in many forms of synaptic plasticity. To assess whether N-methyl-d-aspartate receptors mediate the increase in GluR1 and spine density resulting from cholinergic deafferentation, we examined the effect of N-methyl-d-aspartate receptor blockade on nucleus basalis lesion-induced upregulation of GluR1 and dendritic spines. Rats received unilateral sham or 192 IgG saporin lesions of the nucleus basalis. Half of the rats in each group were treated with the N-methyl-d-aspartate antagonist MK-801 or phosphate-buffered saline. Two weeks later, brains were processed for either immunohistochemical staining of the GluR1 subunit or Golgi histology. In layer II-III of frontal cortex, neuronal GluR1 expression was assessed using an unbiased stereological technique, and spine density was assessed on basilar branches of pyramidal neurons. GluR1 expression was increased after nucleus basalis lesion, but this increase was prevented with MK-801. Similarly, nucleus basalis-lesioned animals had significantly higher spine densities, and this effect was also prevented by treatment with MK-801. Thus, N-methyl-d-aspartate receptor blockade prevented both GluR1 and spine density upregulation following cholinergic deafferentation, suggesting that these effects are N-methyl-d-aspartate receptor-mediated.

Effect of nucleus basalis magnocellularis cholinergic lesions on fear-like and anxiety-like behavior.

Previous research has suggested that cholinergic neurons in the nucleus basalis magnocellularis and substantia innominata (NBM/SI) may be important in mediating aversive states. The authors investigated the effect of NBM/SI cholinergic lesions, induced with 192 IgG saporin, on behavioral measures of aversive states in rats. Behavior in the elevated plus maze and behavioral suppression induced by 2 fear-conditioned stimuli, a tone and a light, were evaluated. Lesions had no effect on any measures in the elevated plus maze but attenuated operant suppression induced by the light and attenuated freezing induced by the tone, though this last effect was not statistically significant. The results of the study suggest that NBM/SI cholinergic neurons may be important in mediating selective aspects of aversive states.

Susceptibility of adult Coccinella septempunctata (Coleoptera: Coccinellidae) to insecticides with different modes of action.

Five insecticides (pyriproxifen, imidacloprid, deltamethrin + heptenophos, lambda-cyhalothrin and Bacillus thuringiensis Berliner subsp. tenebrionis) were examined in the laboratory for their acute detrimental side-effects at field rates on adult seven-spot ladybird beetle, Coccinella septempunctata L. The toxicity of the preparations was determined by measuring the acute surface contact effects (dried spray on leaves of Philadelphus coronarius L.), except for B. thuringiensis where mixed pollen was treated. Four to six concentrations were tested (pyriproxifen 12.5, 25, 50, 100, 200, 400 mg AI litre(-1); imidacloprid 62.4, 125, 250, 500 mg AI litre(-1); deltamethrin + heptenophos 26.4, 53.1, 106.3, 212.5 mg AI litre(-1); lambda-cyhalothrin 1.1, 3.4, 10, 30 mg AI litre(-1); B. thuringiensis 1.5, 3.0, 12.0, 48, 192, 768 mg AI litre(-1)), with 22 adults exposed per concentration. All tests were conducted in the laboratory of the Plant Protection Department (University of Debrecen, Hungary) at 22-25 degrees C, 40-60% RH, under a 16:8 h light:dark photoperiod in 1998-1999. Data were analyzed by probit analysis, probit transformation and analysis of variance. According to different categories of evaluation, pyriproxifen, imidacloprid and B. thuringiensis subsp. tenebrionis seem to be safe for C. septempunctata adults but the other two preparations were moderately harmful to them, which requires further semi-field or field tests to measure their real effect under field conditions.

Pain sensitivity following loss of cholinergic basal forebrain (CBF) neurons in the rat.

Flexion/withdrawal reflexes are attenuated by spinal, intracerebroventricular (ICV) and systemic delivery of cholinergic agonists. In contrast, some affective reactions to pain are suppressed by systemic cholinergic antagonism. Attention to aversive stimulation can be impaired, as is classical conditioning of fear and anxiety to aversive stimuli and psychological activation of stress reactions that exacerbate pain. Thus, in contrast to the suppressive effects of cholinergic agonism on reflexes, pain sensitivity and affective reactions to pain could be attenuated by reduced cerebral cholinergic activation. This possibility was evaluated in the present study, using an operant test of escape from nociceptive thermal stimulation (10 °C and 44.5 °C) before and after destruction of basal forebrain cholinergic neurons. ICV injection of 192 IgG-saporin produced widespread loss of basal forebrain cholinergic innervation of the cerebral cortex and hippocampus. Post-injection, escape from thermal stimulation was decreased with no indication of recovery for upto 19 weeks. Also, the normal hyperalgesic effect of sound stress was absent after ICV 192-sap. Effects of cerebral cholinergic denervation or stress on nociceptive licking and guarding reflexes were not consistent with the effects on operant escape, highlighting the importance of evaluating pain sensitivity of laboratory animals with an operant behavioral test. These results reveal that basal forebrain cholinergic transmission participates in the cerebral processing of pain, which may be relevant to the pain sensitivity of patients with Alzheimer's disease who have prominent degeneration of basal forebrain cholinergic neurons.

Aging and cholinergic deafferentation alter GluR1 expression in rat frontal cortex.

Previously, we demonstrated that plasticity of frontal cortex is altered in aging rats: lesions of the nucleus basalis magnocellularis (NBM) produce larger declines in dendritic morphology in frontal cortex of aged rats compared to young adults. Cholinergic afferents from the NBM modulate glutamatergic transmission in neocortex, and glutamate is known to be involved in dendritic plasticity. To begin to identify possible mechanisms underlying age-related differences in plasticity after NBM lesion, we assessed the effect of cholinergic deafferentation on expression of the AMPA receptor subunit GluR1 in frontal cortex of young adult and aging rats. Young adult, middle-aged, and aged rats received sham or 192 IgG-saporin lesions of the NBM, and an unbiased stereological technique was used to estimate the total number of intensely GluR1-immunopositive neurons in layer II-III of frontal cortex. While the number of GluR1-positive neurons was increased in both middle-aged and aged rats, lesions markedly increased the number of intensely GluR1-immunopositive neurons in frontal cortex of young adult rats only. This age-related difference in lesion-induced expression of AMPA receptor subunit protein could underlie the age-related differences in dendritic plasticity after NBM lesions.