Brain Cholinergic Neurons Research Articles

Cholinergic Modulation of General Anesthesia

Acetylcholine in the brain promotes arousal and facilitates cognitive functions. Cholinergic neurons in the mesopontine brainstem and basal forebrain are important for activation of the cerebral cortex, which is characterized by the suppression of irregular slow waves, an increase in gamma (30-100 Hz) activity in the electroencephalogram, and the appearance of a hippocampal theta rhythm. During general anesthesia, a decrease in acetylcholine release and cholinergic functions contribute to the desired outcomes of general anesthesia, such as amnesia, loss of awareness and consciousness, and immobility. Animal experiments indicate that inactivation, lesion, or genetic ablation of cholinergic neurons in the basal forebrain potentiated the effects of inhalational and injectable anesthetics, including isoflurane, halothane, propofol, pentobarbital, and in some cases, ketamine. Increased behavioral sensitivity to general anesthesia, faster induction time, and delayed recovery of a loss of righting reflex have been observed in rodents with basal forebrain cholinergic deficits. Cholinergic stimulation in the prefrontal cortex, thalamus, and basal forebrain hastens recovery from general anesthesia. Anticholinesterase accelerates emergence from general anesthesia, but with mixed success, in part depending on the anesthetic used. Cholinergic deficits may contribute to cognitive impairments after anesthesia and operations, which are severe in aged subjects. We propose a cholinergic hypothesis for postoperative cognitive disorder, in line with the cholinergic deficits and cognitive decline in aging and Alzheimer’s disease. The current animal literature suggests that brain cholinergic neurons can regulate the immune and inflammatory response after surgical operation and anesthetic exposure, and anticholinesterase and α7-nicotinic cholinergic agonists can alleviate postoperative inflammatory response and cognitive deficits.

(-)-o-[11 C]methyl-trans-decalinvesamicol ((-)-[11 C]OMDV) as a PET ligand for the vesicular acetylcholine transporter.

To develop a PET imaging agent to visualize brain cholinergic neurons and synaptic changes caused by Alzheimer's disease, (-)- and (+)-o-[11 C]methyl-trans-decalinvesamicol ([11 C]OMDV) were isolated and investigated for differences in not only their binding affinity and selectivity to vesicular acetylcholine transporter (VAChT), but also their in vivo activities. [11 C]OMDV has a high binding affinity for VAChT both in vitro and in vivo. Racemic OMDV and o-trimethylstannyl-trans-decalinvesamicol (OTDV), which are precursors for synthesis of [11 C]OMDV, were separated into (-)-optical isomers ((-)-OMDV and (-)-OTDV) and (+)-optical isomers ((+)-OMDV and (+)-OTDV) by HPLC. In the in vitro binding assay, (-)-OMDV(7.2nM) showed eight times higher binding affinity (Ki) to VAChT than that of (+)-OMDV(57.5nM). In the biodistribution study, the blood-brain barrier permeability of both enantiomers ((-)-[11 C]OMDV and (+)-[11 C]OMDV) was similarly high (about 1.0%ID/g) at 2min post-injection. However, (+)-[11 C]OMDV clearance from the brain was faster than (-)-[11 C]OMDV. In the in vivo blocking study, accumulation of (-)-[11 C]OMDV in the cortex was markedly decreased (approximately 30% of control) by coadministration of vesamicol, and brain uptake of (-)-[11 C]OMDV was not significantly altered by coadministration of (+)-pentazocine or (+)-3-(3-hydroxyphenyl)-N-propylpiperidine ((+)-3-PPP). PET-CT imaging revealed inhibition of the rat brain uptake of (-)-[11 C]OMDV by coadministration of vesamicol. In conclusion, (-)-[11 C]OMDV, which is an enantiomer of OMDV, selectively binds to VAChT with high affinity in the rat brain in vivo. (-)-[11 C]OMDV may be utilized as a potential PET ligand for studying presynaptic cholinergic neurons in the brain.

Aggravated effects of coexisting marginal thiamine deficits and zinc excess on SN56 neuronal cells

ABSTRACT Objectives: Zinc excitotoxicity and thiamine pyrophosphate deficiency (TD) are known pathogenic signals contributing to mechanism of different encephalopathies through inhibition of enzymes responsible for energy metabolism such as pyruvate dehydrogenase, aconitase or ketoglutarate dehydrogenase. The aim of this work was to investigate whether subclinical Zn excess and TD, frequent in aging brain, may combine yielding overt neuronal impairment. Results: Clonal SN56 cholinergic neuronal cells of septal origin were used as the model of brain cholinergic neurons, which are particularly susceptible to neurodegeneration in the course of Alzheimer’s disease, hypoxia and other dementia-linked brain pathologies. Neither subtoxic concentration of Zn (0.10 mM) nor mild 20–25% TD deficits alone caused significant negative changes in cultured cholinergic neurons viability and their acetyl-CoA/acetylcholine metabolism. However, cells with mild TD accumulated Zn in excess, which impaired their energy metabolism causing a loss of neurons viability and their function as neurotransmitters. These negative effects of Zn were aggravated by amprolium which is an inhibitor of thiamine intracellular transport. Conclusion: Our data indicate that TD may amplify otherwise non-harmful border-line Zn excitotoxic signals yielding progress of neurodegeneration.

Scopolamine-induced amnesia model: A possible anticholinergic mechanism with reversibility with statins and nootropic agents

Background: High cholesterol turnover is implicated in the pathogenesis of Alzheimer’s disease by causing β amyloid plaque deposition and selective degeneration of cholinergic neurons in brain whereas drugs currently used for the treatment of the same are based on mechanisms which are cholesterol independent. Aims and Objectives: The aim of the study is to find any cholesterol independent mechanisms such as antioxidant and cholinomimetic action of statins in a reversal of memory deficit in scopolamine-induced amnesia. Materials and Methods: Sixty young Swiss albino mice were divided equally into 10 groups, i.e., Group I to Group X. Group I–V and Group VI–X were subjected two exteroceptive behavioral models, i.e., passive avoidance paradigm and Morris water maze, (MWM) respectively. Group I and Group VI served as negative control. Carboxymethyl cellulose (CMC), atorvastatin, simvastatin (test drugs), and piracetam (standard drug), respectively, were given orally to Group II and VII, Group III and VIII, Group IV and IX, and Group V and X for 14 days before inducing amnesia using scopolamine (0.4 mg/kg body weight) injection i.p. Results: Step down latency of disease control group treated with CMC was significantly lower than that of the negative control group (42.5 ± 9.07 s vs. 147.5 ± 10.78 s) as well as that of statin and piracetam treated groups. Learning was good among all groups in MWM with a gradual decrease in escape latency from day 1 to day 4 but during memory retrieval test on day 5, CMC treated group performed poorly in comparison to others. There was no significant difference in plasma cholesterol levels among different groups whereas malondialdehyde was significantly lower among groups treated with statins or piracetam. Conclusion: Statins reversed the scopolamine-induced amnesia by some cholesterol independent mechanisms.

Nerve growth factor-promotes primary cilium assembly in cholinergic neurons from the human basal forebrain

The primary cilium is a non-motile sensory antenna protruding from the surface of nearly all cells of the body, able to mediate the cellular response to extracellular signals. Although many of its functions remain to be clarified, it has been recently shown a role in neurogenesis [1]. In this study we evaluated the presence of the primary cilium in neurons isolated from the human fetal nucleus basalis of Meynert (hfnbM), a basal forebrain region crucially involved in the cholinergic transmission required for learning and memory. The hfnbM cells are characterized by the expression of cholinergic markers, such as choline acetyl transferase (ChAT) and also express the primary cilium, which, in basal conditions, was detected in the 17% of cells. It is known that nerve growth factor (NGF) supports survival, maintenance, connectivity and function of the brain cholinergic neurons. Indeed, we demonstrated that hfnbM cells respond to NGF in terms of proliferation, neurite formation and ChAT expression. Interestingly, NGF exposure significantly increased the percentage of ciliated cells (34.9%±1.8%). Given the known adverse effect of systemic chronic inflammation in the pathogenesis of neurodegenerative diseases, such as Alzheimer’s disease, characterized by the loss of nbM neurons, we exposed our cells to tumor necrosis factor-α (TNF-α). We observed that TNF-α significantly reduced the number of ciliated cells (4.3%±2%). Our results strongly suggest for the first time that primary cilia may be involved in the NGF-driven maturation of human nbM cholinergic neurons and suggest that the deleterious effects of neuroinflammation may be linked to an altered formation of the primary cilium.

Heterozygous disruption of activin receptor-like kinase 1 is associated with increased arterial pressure in mice.

ABSTRACTThe activin receptor-like kinase 1 (ALK-1) is a type I cell-surface receptor for the transforming growth factor-β (TGF-β) family of proteins. Hypertension is related to TGF-β1, because increased TGF-β1 expression is correlated with an elevation in arterial pressure (AP) and TGF-β expression is upregulated by the renin-angiotensin-aldosterone system. The purpose of this study was to assess the role of ALK-1 in regulation of AP using Alk1 haploinsufficient mice (Alk1+/−). We observed that systolic and diastolic AP were significantly higher in Alk1+/− than in Alk1+/+ mice, and all functional and structural cardiac parameters (echocardiography and electrocardiography) were similar in both groups. Alk1+/− mice showed alterations in the circadian rhythm of AP, with higher AP than Alk1+/+ mice during most of the light period. Higher AP in Alk1+/− mice is not a result of a reduction in the NO-dependent vasodilator response or of overactivation of the peripheral renin-angiotensin system. However, intracerebroventricular administration of losartan had a hypotensive effect in Alk1+/− and not in Alk1+/+ mice. Alk1+/− mice showed a greater hypotensive response to the β-adrenergic antagonist atenolol and higher concentrations of epinephrine and norepinephrine in plasma than Alk1+/+ mice. The number of brain cholinergic neurons in the anterior basal forebrain was reduced in Alk1+/− mice. Thus, we concluded that the ALK-1 receptor is involved in the control of AP, and the high AP of Alk1+/− mice is explained mainly by the sympathetic overactivation shown by these animals, which is probably related to the decreased number of cholinergic neurons.

Aging elevates metabolic gene expression in brain cholinergic neurons

The basal forebrain (BF) cholinergic system is selectively vulnerable in human brain diseases, while the cholinergic groups in the upper pons of the brainstem (BS) resist neurodegeneration. Cholinergic neurons (200 per region per animal) were laser-microdissected from five young (8 months) and five aged (24 months) F344 rats from the BF and the BS pontine lateral dorsal tegmental/pedunculopontine nuclei (LDTN/PPN) and their expression profiles were obtained. The bioinformatics program SigPathway was used to identify gene groups and pathways that were selectively affected by aging. In the BF cholinergic system, aging most significantly altered genes involved with a variety of metabolic functions. In contrast, BS cholinergic neuronal age effects included gene groupings related to neuronal plasticity and a broad range of normal cellular functions. Transcription factor GA-binding protein alpha (GABPα), which controls expression of nuclear genes encoding mitochondrial proteins, was more strongly upregulated in the BF cholinergic neurons (+107%) than in the BS cholinergic population (+40%). The results suggest that aging elicits elevates metabolic activity in cholinergic populations and that this occurs to a much greater degree in the BF group than in the BS group.

Streptozotocin-induced diabetes is associated with changes in NGF levels in pancreas and brain.

Type 1 diabetes mellitus (DM), a "classical" result of a pancreatic-beta cell damage, is associated with various metabolic, neuronal, endocrine and immune alterations at cellular, tissue and organ levels. Nerve growth factor (NGF) is one of the most extensively studied neurotrophic factors, which is produced and released by numerous cells including the pancreatic beta cells. NGF plays an important role during brain development and may be able to delay or even reverse damaged forebrain cholinergic neurons that undergo degeneration in aged animals and in Alzheimer's disease (AD). Recent reports indicate that experimentally induced DM in rodents can cause brain biochemical and molecular alterations similar to those observed in sporadic AD. Given the importance of NGF in the pathophysiology of brain cholinergic neurons, we looked for NGF changes in the pancreas and brain of diabetic rats. The aim of this study was, therefore, to investigate the effect of streptozotocin-induced DM on NGF and NGF receptor expression in pancreas and brain. The results showed that DM is associated with altered NGF, NGF-receptor expression in both pancreas and brain.

Effects of acidosis on brain capillary endothelial cells and cholinergic neurons: relevance to vascular dementia and Alzheimer's disease

Alzheimer's disease is a progressive brain disorder which is neuropathologically characterized by an increased number of beta-amyloid plaques, tau pathology and synapse loss. Recent research suggests that vascular pathology may be also important for the development and progression of Alzheimer's disease. It is still unknown whether there is a relation between damage of brain capillary endothelial cells (BCEC) and subsequent cholinergic cell death. The aim of this study was to examine the effects of acidosis on cell death of BCEC and cholinergic neurons in an organotypic brain slice model. We show that BCEC were heavily damaged in medium at pH<6.6. Cholinergic neurons incubated in medium pH 6.0 degenerated within 2–3 days and were not rescued by nerve growth factor (NGF). Lactate did not affect the survival of BCEC or cholinergic neurons. Both BCEC and cholinergic cells were not affected at pH 7.4, 7.0 or 6.6. It is concluded that both endothelial cells and cholinergic neurons have a high capacity to compensate for pH changes. At a certain pH, however, the vascular and neuronal cells show the same vulnerability, indicating that a low pH is deleterious for the cerebral microenvironment. Future studies are necessary to explore whether temporary pH changes could be responsible for cerebrovascular damage and cholinergic cell death. Acidosis may play an important role in the development of vascular dementia and Alzheimer's disease.

82-kDa choline acetyltransferase is in nuclei of cholinergic neurons in human CNS and altered in aging and Alzheimer disease

Cholinergic neurons express choline acetyltransferase (ChAT) which synthesizes acetylcholine. We show here for the first time that primate-specific 82-kDa ChAT is expressed in nuclei of cholinergic neurons in human brain and spinal cord; isoform-specific antibodies were used to compare localization patterns and temporal expression of the more abundant 69-kDa ChAT and primate-specific 82-kDa ChAT in necropsy tissues. The 82-kDa ChAT co-localizes with 69-kDa ChAT in well-characterized cholinergic areas, but is also found in the claustrum which does not contain 69-kDa ChAT. Cholinergic neuron function changes with increasing age and are targeted in neurodegenerative diseases such as AD, thus we compared expression and subcellular localization of 69- and 82-kDa ChAT in necropsy brain samples from control subjects of varying ages and from Alzheimer disease (AD) subjects. The 82-kDa ChAT protein was expressed in cholinergic neurons in brain from birth until the eighth decade of life and in AD, but the subcellular staining pattern and proportion of neurons that were immunopositive changed with increasing age and in AD.

Phenotype-dependent susceptibility of cholinergic neuroblastoma cells to neurotoxic inputs

A preferential loss of brain cholinergic neurons in the course of Alzheimer's disease and other encephalopathies is accompanied by a proportional impairment of acetyl-CoA synthesizing capacity in affected brains. Particular susceptibility of cholinergic neurons to neurodegeneration might results from insufficient supply of acetyl-CoA for energy production and acetylcholine synthesis in these conditions. Exposure of SN56 cholinergic neuroblastoma cells to dibutyryl cAMP and retinoic acid for 3 days caused their morphologic differentiation along with the increase in choline acetyltransferase activity, acetylcholine content and release, calcium content, and the expression of p75 neurotrophin receptors. Acetyl-CoA content correlated inversely with choline acetyltransferase activity in different lines of SN56 cells. In differentiated cells, aluminum (1 mM), amyloid beta(25-35) (0.001 mM), and sodium nitroprusside (1 mM), caused much greater decrease of pyruvate dehydrogenase and choline acetyltransferase activities and cell viability than in nondifferentiated ones. Aluminum (1 mM) aggravated suppressory effects of amyloid beta on choline acetyltransferase and pyruvate dehydrogenase activities and viability of differentiated cells. Similar additive inhibitory effects were observed upon combined exposure of differentiated cells to sodium nitroprusside and amyloid beta(25-35). None or much smaller suppressory effects of these neurotoxins were observed in nondifferentiated cells. Increase in the fraction of nonviable differentiated cells positively correlated with losses of choline acetyltransferase, pyruvate dehydrogenase activities, and cytoplasmic cytochrome c content in different neurotoxic conditions. These data indicate that highly differentiated cholinergic neurons may be more susceptible to aluminum and other neurotoxins than the nondifferentiated ones due to relative shortage of acetyl-CoA, increased content of Ca(2+), and expression of p75 receptors, yielding increase in cytoplasmic cytochrome c and subsequently grater rate of death of the former ones.

A model for dynamic regulation of choline acetyltransferase by phosphorylation

Choline acetyltransferase (ChAT) synthesizes the neurotransmitter acetylcholine (ACh) and is a phenotypic marker for cholinergic neurons. Cholinergic neurons in brain are involved in cognitive function, attentional processing and motor control, and decreased ChAT activity is found in several neurological disorders including Alzheimer's disease. Dysregulation of ChAT and cholinergic communication is also associated with some spontaneous point-mutations in ChAT that alter its substrate binding kinetics, or by disruption of signaling pathways that could regulate protein kinases for which ChAT is a substrate. It has been identified recently that the catalytic activity and subcellular distribution of ChAT, and its interaction with other cellular proteins, can be modified by phosphorylation of the enzyme by protein kinase-C and Ca2+/calmodulin-dependent protein kinase II; these kinases appear also to mediate some of the effects of beta-amyloid peptides on cholinergic neuron functions, including the effects on ChAT. This review outlines a new model for the regulation of cholinergic transmission at the level of the presynaptic terminal that is mediated by hierarchically-regulated, multi-site phosphorylation of ChAT.

The effect of aging on the subcellular distribution of estrogen receptor‐alpha in the cholinergic neurons of transgenic and wild‐type mice

The degeneration of the basal forebrain cholinergic system plays an important role in cognitive deterioration in aging and Alzheimer's disease. Brain cholinergic neurons and their projections are affected by changes in the circulating levels of estrogens, which exert their effects mainly through the estrogen receptors. In this study, we investigated the effect of aging, estrogen status and transgenic genotype on the number of cholinergic neurons and the estrogen receptor alpha (ERalpha) content in the medial septum-vertical limb of the diagonal band of Broca. We used 6- and 12-month-old female double transgenic mice carrying mutated human amyloid precursor protein (APPswe) and presenilin-1 (PS1-A246E), and their nontransgenic littermate controls, which had been sham-operated or ovariectomized at the age of 3 months. Brain sections were double immunostained for choline acetyltransferase (ChAT) and ERalpha and used for stereological cell counting. We found that the number of ChAT-immunoreactive (ir) neurons containing nuclear ERalpha-ir was significantly lower in 12- than in 6-month-old mice. However, the age of the mice, the transgenic genotype or ovariectomy had no effect on the total number of ChAT-ir neurons, or on the number and percentage of all ChAT-ir neurons that contained ERalpha. These results indicate that aging is associated with translocation of ERalphas from the nucleus to the cytoplasm. We propose that this phenomenon is linked to those age-related processes known to be involved in inhibiting ERalpha binding to nuclei.

Nerve growth factor and acetyl‐L‐carnitine evoked shifts in acetyl‐CoA and cholinergic SN56 cell vulnerability to neurotoxic inputs

Different groups of brain cholinergic neurons display variable susceptibility to similar neurotoxic inputs. The aim of this work was to find out whether changes in cholinergic phenotype may alter the availability of acetyl-CoA in mitochondrial compartment and thereby the viability of cholinergic neurons. Cyclic AMP (cAMP) and retinoic acid caused differentiation (DC) of T17 TrkA(+) cholinergic neuroblastoma cells. In addition, it increased the choline acetyltransferase (ChAT) activity, Ca(2+) accumulation and cytoplasmic acetyl-CoA level, but decreased mitochondrial acetyl-CoA and cell resistance to amyloid-beta(25-35) (Abeta) toxicity. Nerve growth factor (NGF) caused similar alterations in the nondifferentiated cells (NC). On the other hand, in DC NGF suppressed ChAT activity and elevated mitochondrial level of acetyl-CoA but also caused a further increase of Ca(2+) content and cell susceptibility to Abeta. The significant inverse correlation was found between ChAT activity and mitochondrial levels of acetyl-CoA. Abeta markedly reduced the expression of cholinergic phenotype, acetyl-CoA content, and viability of DC. These effects were absent or much less pronounced in NC. Acetyl-L-carnitine reversed suppressing effects of Abeta on acetyl-CoA levels and ChAT activity but did not reverse increased mortality in DC. Presented data indicate that increased transmitter activity in highly differentiated cholinergic neurons, decreased acetyl-CoA level in their mitochondrial compartment, and increased Ca(2+) accumulation can make them more prone to neurotoxic conditions. Phenotype-dependent changes in intracellular distribution of acetyl-CoA thus play an important role in regulation of viability and transmitter function in brain cholinergic neurons.

Memory consolidation and reconsolidation of an inhibitory avoidance response in mice: effects of i.c.v. injections of hemicholinium-3

The immediate post-training i.c.v. administration of hemicholinium-3 (HC-3) (1 μg), a specific inhibitor of the high-affinity choline uptake (HACU) in brain cholinergic neurons, impaired retention test performance of a one-trial step-through inhibitory avoidance response in adult male CF-1 mice. The effect was observed in mice that received a footshock (0.8 mA, 50 Hz, 1 s) on the learning trial, and not only 48 h after training, but also 7 days after it. After the completion of the retention test at each of the training-test interval that were studied, the HACU in the hippocampus of HC-3-treated mice was not significantly different from that of saline-injected (1 μl) control groups. Mice that were over-reinforced (1.2 mA, 50 Hz, 1 s) on the learning trial, exhibited a high retention performance 48 h after training. The immediate i.c.v. injection of HC-3 (1 μg) after the retention test, that is, after memory reactivation, significantly impaired retention performance over 4 consecutive days, whereas the saline-injected control group shown a slight, but significant performance decrease only at the last retention test. Retention performance was unchanged in HC-3-treated mice not undergoing memory reactivation session. These results, taken together, indicate that HC-3, not only impaired consolidation, but also reconsolidation of an inhibitory avoidance task in mice, suggesting a critical participation of central cholinergic mechanisms in both memory processes.

Brain microdialysis study of the effects of hazardous chemicals on the central nervous system 2. Toluene exposure and cerebral acetylcholine.

The microdialysis technique was applied to detect the changes in the activity of acetylcholine (ACh) neurons in the rat brain. The effects of intraperitoneal (i.p.) injection of toluene on the amount of ACh release from the nerve terminals of the brain cholinergic neurons were investigated in freely moving rats. In the striatum, injection of toluene decreased the extracellular concentration of ACh in a dose dependent manner in the range 200 to 2,000 mg/kg. Similar effects of toluene on ACh release were observed in the hippocampus after i.p. administration. The increases in ACh content in brain homogenate after i.p. injection of toluene seemed to be caused by the decreased release of ACh from cholinergic nerve endings. Injection of toluene at doses higher than 200 mg/kg decreased ACh release and a similar decrease was suggested to occur in 8-h inhalation exposure to toluene at 1,000 ppm or higher concentrations.

Co-expression of α7 and β2 nicotinic acetylcholine receptor subunit mRNAs within rat brain cholinergic neurons

Nicotine enhances cognitive and attentional processes through stimulation of the basal forebrain cholinergic system. Although muscarinic cholinergic autoreceptors have been well characterized, pharmacological characterization of nicotinic autoreceptors has proven more difficult. The present study used double-labeling in situ hybridization to determine expression of nicotinic acetylcholine receptor (nAChR) subunit mRNAs within basal forebrain cholinergic neurons in order to gain information about possible nAChR autoreceptor properties. Cholinergic cells of the mesopontine tegmentum and striatal interneurons were also examined, as were septohippocampal GABAergic neurons that interact with cholinergic neurons to regulate hippocampal activity. α7 and β2 nAChR mRNAs were found to be co-expressed in almost all cholinergic cells and in the majority of GABAergic neurons examined. α4 nAChR mRNA expression was restricted to cholinergic cells of the nucleus basalis magnocellularis, and to non-cholinergic cells of the medial septum and mesopontine tegmentum. These data suggest possible regional differences in the pharmacological properties of nicotinic autoreceptors on cholinergic cells. Whereas most cholinergic cells express rapidly desensitizing α7 homomers or α7β2 heteromers, cortical projection neurons may also express a pharmacologically distinct α4β2 nAChR subtype. There may also be differential nAChR regulation of cholinergic and non-cholinergic cells within the mesopontine tegmentum that are implicated in acquisition of nicotine self-administration.

Differential toxicity of nitric oxide, aluminum, and amyloid β-peptide in SN56 cholinergic cells from mouse septum

A characteristic feature of several encephalopathies is preferential impairment of cholinergic neurons. Their particular susceptibility to cytotoxic insults may result from the fact that they utilise acetyl-CoA both for energy production and acetylcholine synthesis. In addition, phenotypic modifications of cholinergic neurons are likely to influence their susceptibility to specific harmful conditions. SN56 cholinergic cells were differentiated by the combination of dibutyryl cAMP and retinoic acid. Al and sodium nitroprusside (SNP, NO donor) exerted direct additive inhibitory effects on mitochondrial aconitase activity. However, NO, Al, or amyloid β (Aβ)(25–35) caused none or only slight changes of choline O-acetyl transferase (ChAT) and pyruvate dehydrogenase (PDH) activity and relatively small loss of non-differentiated cells (NCs). On the other hand, in differentiated cells (DCs) these neurotoxins brought about marked decreases of these enzyme activities along with greater than in non-differentiated ones increase of cell-death rate. Aβ(35–25) had no effect on these cell parameters. NO and other compounds aggravated detrimental effect of each other particularly in differentiated cells. Thus, differential vulnerability of brain cholinergic neurons to various degenerative signals may result from their phenotype-dependent ratios of acetylcholine to acetyl-CoA synthesising capacities.

Aluminum, NO, and nerve growth factor neurotoxicity in cholinergic neurons.

Several neurotoxic compounds, including Al, NO, and beta-amyloid may contribute to the impairment or loss of brain cholinergic neurons in the course of various neurodegenerative diseases. Genotype and phenotypic modifications of cholinergic neurons may determine their variable functional competency and susceptibility to reported neurotoxic insults. Hybrid, immortalized SN56 cholinergic cells from mouse septum may serve as a model for in vitro cholinotoxicity studies. Differentiation by various combinations of cAMP, retinoic acid, and nerve growth factor may provide cells of different morphologic maturity as well as activities of acetylcholine and acetyl-CoA metabolism. In general, differentiated cells appear to be more susceptible to neurotoxic signals than the non-differentiated ones, as evidenced by loss of sprouting and connectivity, decreases in choline acetyltransferase and pyruvate dehydrogenase activities, disturbances in acetyl-CoA compartmentation and metabolism, insufficient or excessive acetylcholine release, as well as increased expression of apoptosis markers. Each neurotoxin impaired both acetylcholine and acetyl-CoA metabolism of these cells. Activation of p75 or trkA receptors made either acetyl-CoA or cholinergic metabolism more susceptible to neurotoxic influences, respectively. Neurotoxins aggravated detrimental effects of each other, particularly in differentiated cells. Thus brain cholinergic neurons might display a differential susceptibility to Al and other neurotoxins depending on their genotype or phenotype-dependent variability of the cholinergic and acetyl-CoA metabolism.

Beta -Amyloid peptide blocks the response of alpha 7-containing nicotinic receptors on hippocampal neurons.

Alzheimer's disease produces a devastating decline in mental function, with profound effects on learning and memory. Early consequences of the disease include the specific loss of cholinergic neurons in brain, diminished cholinergic signaling, and the accumulation of beta-amyloid peptide in neuritic plaques. Of the nicotinic acetylcholine receptors at risk, the most critical may be those containing the alpha7 gene product (alpha7-nAChRs), because they are widespread, have a high relative permeability to calcium, and regulate numerous cellular events in the nervous system. With the use of whole-cell patch-clamp recording we show here that nanomolar concentrations of beta-amyloid peptides specifically and reversibly block alpha7-nAChRs on rat hippocampal neurons in culture. The block is noncompetitive, voltage-independent, and use-independent and is mediated through the N-terminal extracellular domain of the receptor. It does not appear to require either calcium influx or G protein activation. beta-Amyloid blockade is likely to be a common feature of alpha7-nAChRs because it applies to the receptors at both somato-dendritic and presynaptic locations on rat hippocampal neurons and extends to homologous receptors on chick ciliary ganglion neurons as well. Because alpha7-nAChRs in the central nervous system are thought to have numerous functions and recently have been implicated in learning and memory, impaired receptor function in this case may contribute to cognitive deficits associated with Alzheimer's disease.