Choline Acetyltransferase Concentrations Research Articles

Cognitive impairment induced by sevoflurane anesthesia is mediated by the cholinergic system after gastrointestinal surgery in older patients: A randomized, controlled trial.

Delayed neurocognitive recovery after surgery is associated with increased morbidity and mortality. However, its mechanism of action remains controversial and complex.A prospective, double-blind, randomized controlled trial was performed at the Affiliated Hospital of Zunyi Medical University. Older patients (aged 65 years and older) who underwent gastrointestinal surgery were randomly divided into sevoflurane-based or propofol-based anesthesia groups. The Mini-Mental State Examination was performed to evaluate cognitive function. Peripheral venous blood was collected to test the levels of choline acetyltransferase and acetylcholinesterase. A total of 75 patients were enrolled and 30 patients in each group completed the study. On Day 1 postoperation, patients in the sevoflurane group showed worse performance on the Mini-Mental State Examination than patients in the propofol group. Lower blood choline acetyltransferase concentrations and higher acetylcholinesterase concentrations were observed in patients who had sevoflurane anesthesia than in patients who had propofol anesthesia 1 day postoperative. At 3 days postoperation, patients with sevoflurane- or propofol-based general anesthesia did not differ regardless of Mini-Mental State Examination score or choline acetyltransferase and acetylcholinesterase levels. Sevoflurane-based anesthesia has short-term delayed neurocognitive recovery in older surgical patients, which may be related to central cholinergic system degeneration.

Efficiency of donepezil in elderly patients undergoing orthopaedic surgery due to underlying post-operative cognitive dysfunction: study protocol for a multicentre randomised controlled trial

BackgroundPost-operative cognitive dysfunction (POCD) is an overarching term used to describe cognitive impairment identified in the preoperative or post-operative period. After surgical operations, older patients are particularly vulnerable to memory disturbances and other types of cognitive impairment. However, the pathogenesis of POCD remains unclear with no confirmed preventable or treatable strategy available. Our previous study demonstrated that the concentration of choline acetyl transferase in the cerebral spinal fluid was a predictive factor of POCD and that donepezil, which is an acetylcholinesterase inhibitor used in clinical settings for the treatment of Alzheimer’s disease, can prevent learning and memory impairment after anaesthesia/surgery in aged mice. This study aimed to determine the critical role of donepezil in preventing cognitive impairment in elderly patients undergoing orthopaedic surgery.MethodsA multicentre, double-blind, placebo-controlled, crossover clinical trial will be performed to assess the efficacy of donepezil in elderly patients undergoing orthopaedic surgery. Participants (n = 360) will receive donepezil (5 mg once daily) or placebo from 1 day prior to surgery until 5 days after surgery. Neuropsychological tests will be measured at 1 day before the operation and 1 week, 1 month, 6 months and 1 year after the operation.DiscussionThis research project mainly aimed to study the effects of donepezil in elderly patients undergoing orthopaedic surgery due to underlying POCD and to investigate the underlying physiological and neurobiological mechanisms of these effects. The results may provide important implications for the development of effective interfering strategies, specifically regarding cognitive dysfunction therapy using drugs.Trial registrationClinicalTrials.govNCT04423276. Registered on 14 June 2020

Protective roles of isoastilbin against Alzheimer's disease via Nrf2‑mediated antioxidation and anti‑apoptosis.

By analyzing the L-glutamic acid (L-Glu)-induced apoptosis of PC12 cells and an AlCl3 combined with D-galactose (D-gal)-developed Alzheimer's disease (AD) mouse model, the protective effects of isoastilbin (IAB) against AD were systematically investigated in the present study. Pre-incubation with IAB for 3 h prior to treatment with 25 mM L-Glu decreased cell viability and inhibited apoptosis, suppressed the accumulation of intracellular reactive oxygen species, and restored mitochondrial membrane potential in PC12 cells induced by L-Glu. In mice with AD, the reduced escape latency time in the water maze test, suppressed chronic movement in the center area of an open field test and enhanced ability to seek hidden food in a Y maze test indicated that abnormal behaviors had improved after 28 days of treatment with IAB. Furthermore, IAB reduced the deposition of amyloid β (Aβ) and the expression of phosphorylated-Tau in the mouse brain and enhanced the serum levels of Aβ. IAB ameliorated the oxidative stress via modulating the levels of associated enzymes and improved the functioning of the central cholinergic system, as indicated by an increase in acetylcholine and choline acetyltransferase concentrations. The expression levels of acetylcholine esterase were reduced in the mouse brain in response to IAB pre-treatment. In cells and brain tissue, IAB regulated the expression levels of pro- and anti-apoptotic proteins and enhanced the nuclear levels of NF-E2p45-related factor 2 (Nrf2); subsequently, IAB further enhanced the expression of superoxide dismutase 1, catalase, and heme oxygenase-1 and -2. The findings of the present study indicated that the protection of IAB against AD is at least partially associated with its antioxidation and anti-apoptotic properties.

Protective roles of Amanita caesarea polysaccharides against Alzheimer's disease via Nrf2 pathway

This study explores the neuro-protective effects of Amanita caesarea polysaccharides (ACPS), obtained by 80% alcohol precipitation of water extract and purified using a DEAE-52 cellulose anion exchange column, related to antioxidant activity. A 3-h pre-treatment of ACPS prior to l‑glutamic acid (l‑Glu) co-exposure reversed the decreased cell viability, inhibited apoptosis, suppressed the accumulation of intracellular reactive oxygen species and restored mitochondrial membrane potential in HT22 cells. Compared to l‑Glu-exposed cells, ACPS enhanced the nuclear levels of NF-E2p45-related factor 2 (Nrf2), reduced the cytoplasmic levels of Nrf2 and cytochrome C, suppressed the expression of Kelch-like ECH-associated protein 1, and enhanced the expression of heme oxygenase‑1, superoxide dismutase 1 and cysteine ligase catalytic subunit. In a d‑galactose and aluminum trichloride Alzheimer's disease (AD) mouse model, 42-day administration of ACPS improved the abnormal behaviors. ACPS suppressed the deposition of β‑amyloid peptide in the brain and ameliorated oxidative stress via modulating the levels of related enzymes. ACPS improved the functioning of the central cholinergic system, as indicated by an increase in acetylcholine and choline acetyltransferase concentrations, and reduced acetylcholine esterase levels in the serum, hypothalamus and cerebral cortex. Our data suggest that ACPS may be a promising candidate for the treatment of AD.

Formation of cholinergic synapse-like specializations at developing murine muscle spindles

Muscle spindles are complex stretch-sensitive mechanoreceptors. They consist of specialized skeletal muscle fibers, called intrafusal fibers, which are innervated in the central (equatorial) region by afferent sensory axons and in both polar regions by efferent γ-motoneurons. We show that AChRs are concentrated at the γ-motoneuron endplate as well as in the equatorial region where they colocalize with the sensory nerve ending. In addition to the AChRs, the contact site between sensory nerve ending and intrafusal muscle fiber contains a high concentration of choline acetyltransferase, vesicular acetylcholine transporter and the AChR-associated protein rapsyn. Moreover, bassoon, a component of the presynaptic cytomatrix involved in synaptic vesicle exocytosis, is present in γ-motoneuron endplates but also in the sensory nerve terminal. Finally, we demonstrate that during postnatal development of the γ-motoneuron endplate, the AChR subunit stoichiometry changes from the γ-subunit-containing fetal AChRs to the ε-subunit-containing adult AChRs, similar and approximately in parallel to the postnatal subunit maturation at the neuromuscular junction. In contrast, despite the onset of ε-subunit expression during postnatal development the γ-subunit remains detectable in the equatorial region by subunit-specific antibodies as well as by analysis of muscle spindles from mice with genetically-labeled AChR γ-subunits. These results demonstrate an unusual maturation of the AChR subunit composition at the annulospiral endings and suggest that in addition to the recently described glutamatergic secretory system, the sensory nerve terminals are also specialized for cholinergic synaptic transmission, synaptic vesicle storage and exocytosis.

Reappraising neurotransmitter-based strategies

A number of observations support the hypothesis that a central deficit in acetylcholine (ACh) may be responsible for the initiation of Alzheimer's disease (AD). For example, cholinergic innervation in AD is reduced in areas of the brain important for processing information. Further, reduced concentrations of choline acetyltransferase (ChAT), the enzyme responsible for the synthesis of ACh, correlate with the number of β-amyloid senile plaques and cognitive dysfunction in AD patients. Consequently, several strategies to increase cholinergic neurotransmission have been developed, including ACh precursors, ACh release enhancers, cholinesterase (ChE) inhibitors and receptor agonists. Although ChE inhibitors appear to be the most promising, tacrine, the first ChE inhibitor to be registered and approved for the treatment of AD, has significant tolerability problems. Thus, ChE inhibitors with improved side-effect profiles have been developed and subsequently awarded marketing approval. However, in addition to the cholinergic system that is the most severely affected neurotransmitter system in AD, other neurotransmitter systems may be involved (e.g. serotonergic, noradrenergic and glutamatergic). Therefore, bifunctional compounds or combinations of drugs may provide additional therapeutic value.

Inhibition of dopamine and choline acetyltransferase concentrations in rat CNS neurons by rat α1‐ and α2‐macroglobulins

Inhibition of dopamine and choline acetyltransferase concentrations in rat CNS neurons by rat α1‐ and α2‐macroglobulins

Inhibition of dopamine and choline acetyltransferase concentrations in rat CNS neurons by rat alpha 1- and alpha 2-macroglobulins.

Previous studies have implicated human alpha-2-macroglobulin (alpha2M) as a potential regulator of neuronal development and function. Rat alpha-1-macroglobulin (alpha1M) and acute-phase alpha-2-macroglobulin (alpha2M) are murine homologues of human alpha2M. In this report, we tested the effect of intracranially infused serotonin-activated rat alpha1M (5HT-alpha1M) on the concentration of dopamine (DA) in the corpus striatum in vivo and the effect of 5HT-activated rat alpha1M and alpha2M on the choline acetyltransferase (ChAT) activity upon embryonic basal forebrain neurons in culture. The results show that direct infusion of 0.65 nmole rat 5HT-alpha1M into the adult rat corpus striatum produced a consistent attenuation upon striatal DA concentrations. This decrease was particularly prominent at 5-7 days post-infusion. In addition, rat 5HT-alpha1M and rat 5HT-alpha2M, like human 5HT-alpha2M, all significantly inhibited ChAT activity of embryonic rat cerebral cortex neurons. Although normal human alpha2M and rat alpha2M were either marginally or insignificantly inhibitory in this preparation, normal rat alpha1M dose-dependently inhibited ChAT activity. These results demonstrate that monoamine-activated alpha-macroglobulins from rat depress dopaminergic and cholinergic neurotransmitter systems in the CNS, and this suggests a potential regulatory role of these alpha-macroglobulins in neurotransmitter metabolism.

Neurochemical Effects of Consumption of Great Lakes Salmon by Rats

This study, part of a larger project to determine the health consequences of both perinatal and adult exposure to contaminated salmon from the Great Lakes, determined the neurochemical effects of exposure of rats to chow adulterated with lyophilized salmon fillets. Concentrations of biogenic amines, their metabolites, and choline acetyltransferase (ChAT) were determined in the frontal cortex (FC), nucleus accumbens, caudate nucleus (CN), hippocampus (HC), and substantia nigra (SN) of adult rats who had been exposed, both perinatally and as adults, to standard rat chow adulterated with either 5 or 20% (w/w) lyophilized fillets from either Lake Huron (LH) or Lake Ontario (LO) salmon. Dopamine (DA) concentrations in the FC were significantly decreased following exposure to both 20% fish diets. CN DA concentrations were significantly reduced in rats exposed to all diets, while SN DA was decreased only in the LO20-fed animals. SN norepinephrine concentrations were reduced in all groups except for the LO5-fed rats. 3,4-Dihydroxyphenylacetic acid (DOPAC) concentrations in the FC were significantly increased in the LH20 and LO5 groups, while CN DOPAC concentrations were reduced in LH20, LO5, and LO20 animals. 5-Hydroxyindoleacetic acid concentrations were reduced in the FC and CN of all animals exposed to diets adulterated with Great Lakes salmon. ChAT concentrations were unaffected in rats exposed to any of the adulterated diets. The significant reductions in DA, particularly in the FC and CN, suggest that either fish-borne contaminants or consumption of fish, per se, may affect behaviors that require inhibition of normal responding. We conclude that consumption of contaminated fish from the Great Lakes may result in sufficient reductions in biogenic amine function to result in significant deficits in important behavioral functions in the rat and, by inference, in the perinatally exposed human.

Decreased mesopontine choline acetyltransferase levels in schizophrenia. Correlations with cognitive functions.

The objective was to replicate a reported decrease of choline acetyltransferase (ChAT) in the mesopontine tegmentum of deceased schizophrenics and to see if such a decrease is related to their cognitive status as measured during life. Rigorous antemortem psychiatric evaluations were performed on our large population of schizophrenic patients. Mesopontine tissue was collected promptly following death from eight of these patients, from an additional five schizophrenics without systematic premortem psychiatric evaluation, and from control subjects. ChAT content of this brain tissue was determined using Western immunoblot analysis. There were 13 schizophrenic patients and 8 control subjects. The mean age of subjects in the two groups was similar (64 +/- 9 yr vs 63 +/- 10 yr). Even in the face of reduced post mortem intervals in the patients with schizophrenia, mesopontine tegmental ChAT concentrations were depressed by 70% in schizophrenic patients (1.28 +/- 1.74 vs 4.39 +/- 3.20 ng ChAT/micrograms tissue protein, P < 0.01), and correlated with orientation and reasoning (rs = 0.90 and 0.98, respectively) in those subjects assessed antemortem. Mesopontine ChAT concentrations are depressed in schizophrenia and correlate significantly with measures of cognitive performance in patients with this disorder.

Choline acetyltransferase in schizophrenia

To test the hypothesis that schizophrenia involves altered cholinergic tone in the pons, the authors studied post-mortem brain tissue from subjects with schizophrenia. The authors used Western immunoblot to measure the concentration of choline acetyltransferase, an acetylcholine synthesizing enzyme, in the post-mortem brain tissue of 25 schizophrenic subjects and 28 nonschizophrenic comparison subjects. They also measured the concentration of glial fibrillary acidic protein, a protein from astrocytes, to examine the question of neurodegeneration. The pontine choline acetyltransferase concentrations of the schizophrenic subjects were 46% lower than those of comparison subjects, a significant difference. Glial fibrillary acidic protein concentrations did not differ between the two groups. The lower concentration of choline acetyltransferase in the pontine tegmentum of schizophrenic subjects compared with comparison subjects suggests involvement of pontine cholinergic neurons in schizophrenia.

Preservation of Somatostatin Receptors Coupled to the Inhibition of Adenylate Cyclase in the Cortex and Hippocampus in Senile Dementia of the Alzheimer Type

The distribution and the density of &lt;sup&gt;125&lt;/sup&gt;I-Tyr0-DTrp8-somatostatin (&lt;sup&gt;125&lt;/sup&gt;I-SRIF) binding sites were compared by quantitative radioautography in the hippocampi and entorhinal cortices from normal humans and patients with senile dementia of the Alzheimer type (SDAT). In SDAT, entorhinal cortex concentrations of choline acetyl transferase and SRIF were significantly decreased as compared to controls. The neuroanatomical distribution of &lt;sup&gt;I25&lt;/sup&gt;I-SRIF binding sites was similar in control and SDAT hippocampus and entorhinal cortex, being high in molecular layers of the dentate gyrus and stratum oriens of the CA1 and deeper cortical layers (V-VI), intermediate in superficial (II—III) cortical layers and low in subiculum, hilus of the dentate gyrus and cortical layer I and IV. Competition curves with SMS 201995 to displace &lt;sup&gt;125&lt;/sup&gt;I-SRIF binding allowed to differentiate between high (SS-A)-and low (SS-B)-affinity somatostatin binding sites. In control brains, SS A (K&lt;sub&gt;d&lt;/sub&gt; A = 0.06 ± 0.02 n&lt;i&gt;M&lt;/i&gt;) represented 23–33% of the binding while SS B &lt;i&gt;(&lt;/i&gt;K&lt;sub&gt;d&lt;/sub&gt;B = 23.5 ± 2.8 n&lt;i&gt;M&lt;/i&gt;) accounted for 67–77% of total specific binding, depending on the regions. In dentate gyrus and CA1, the proportion of SS-A was significantly reduced in SDAT as compared to controls. The reduction did not reach significance in the entorhinal cortex, and inhibition of adenylate cyclase activity by SRIF was equivalent in SDAT and controls. It is concluded from these data that the number of SS-A decreases in SDAT hippocampus but that SS-B and negative coupling to adenylate cyclase are preserved.

Transient supersensitivity in the hypertrophied rat urinary bladder

The presence of a paraffin bolus intravesically for 1 or 4 weeks was accompanied by an approximate three-fold increase in bladder weight. A supersensitivity to methacholine was demonstrated in these hypertrophied bladders at 1 week but not at 4 weeks. At the earlier time of observation, the contractile responses in vitro to nerve stimulation were reduced, while at the later time of observation the responses were the same as those of controls; the atropine-sensitive fraction of the response was entirely responsible for the recovery. The concentration of choline acetyltransferase, indicating cholinergic nerve density and initially reduced by about 50%, rose in course of time. It is suggested that in the beginning of the experimental period, the muscle cells of the hypertrophied bladders in situ are exposed to 'subnormal' transmitter concentration. An extension of the field of innervation of the cholinergic nerves may explain the disappearance of the supersensitivity.

Regional tyrosine hydroxylase and choline acetyltransferase concentrations in the brains of lymphocytic choriomeningitis-infected mice

The neurotransmitter biosynthesis enzymes tyrosine hydroxylase and choline acetyltransferase were investigated in selected brain areas of Nya: NYLAR mice infected with lymphocytic choriomeningitis (LCM) virus. Statistically significant alterations in the concentrations of both enzymes occurred in the olfactory, caudate, and neocortical regions at 5 days postinfection. No such alterations occurred in mice given cytoxan (150 mg/kg) 3 days postinfection and examined 5 days postinfection. At 10 days postinfection, however, the cytoxan-treated animals had significantly altered enzyme concentrations in the olfactory region, though not in the caudate or neocortex. This alteration appeared to be transitory, since it was not found in cytoxan-treated animals 60 days postinfection. A possible explanation is that virus production or interference in a brain region cycles over a period of hours or days. Still undetermined is whether these neurochemical changes are a primary effect of the virus or a secondary effect due to the immune response. It is noteworthy that cytoxan caused a marked increase in the enzyme activities studied in most of the brain areas.

Effect of surgical deafferentation of the supraoptic nucleus on its choline acetyltransferase content

The concentration of choline acetyltransferase, a specific marker for cholinergic neurons, was determined in the supraoptic nucleus after a variety of lesions. Surgical lesions immediately rostral as well as medial and lateral to the nucleus did not affect the concentration of the enzyme. Only lesions which separated the nucleus from the posterior part of the lateral hypothalamus slightly decreased its concentration in choline acetyltransferase. It is concluded that the bulk of the cholinergic neurons is in the supraoptic nucleus or its immediate vicinity.

Immunochemical demonstration of increased choline acetyltransferase concentration in rat preoptic area after estradiol administration

Immunochemical demonstration of increased choline acetyltransferase concentration in rat preoptic area after estradiol administration

Aspects of the chemical embryology of the electromotor system of Torpedo marmorata with special reference to synaptogenesis

The functional development of the cholinergic electromotor system of Torpedo marmorata has been followed by measuring four proteins, choline acetyltransferase, acetylcholinesterase, myosin and nicotinic acetylcholine receptor, and the discharge of the electric organ in response to presynaptic stimulation in embryos from 28mm in length to new-born (~ 130 mm). The first two, as indicators of cholinergic function, were followed, both in the electric lobes, sites of the perikarya of the electromotor neurones, and in the electric organ; the last three, indicative respectively of the myofibrillar structures of the myotubes which are the precursors of the electrocytes, the cholinoceptive function of the differentiating postsynaptic membrane, and the existence of functional synapses, were followed in the electric organ. A fall in the tissue concentration of myosin heavy chain protein in the electric organ between 37 ± 2 and 57 ± 2 mm of embryo length correlated well with the transition from vertically orientated myotubes to horizontally flattened electrocytes, with loss in myofibrillar structure. The finding that receptor protein had reached almost 100% of the adult value at an embryo length at which discharges of the electric organ were barely detectable showed that synaptogenesis, seen morphologically as a continuous process commencing with the ingrowth of axons at 55 mm, occurs functionally in two stages in which the formation of a receptor- (and esterase-) rich postsynaptic membrane precedes that of transmitting synapses. Choline acetyltransferase activity was first detected in the lobe at about 40mm embryo length, 4–6 weeks after the electromotor perikarya had stopped dividing. It rose steadily to 125% of the adult value at birth. By contrast the rise in the electric organ was much slower; it reached only 50% of the adult value at birth and showed evidence of two phases, an initial small rise coincident with and perhaps due to the invasion of the electric organ by cholinergic electromotor axons and a more pronounced one from 90 mm accompanied by the formation of transmitting synapses. Significant (1 V) discharges of electric organs in response to single stimuli were first detected at 90mm and rose exponentially to reach adult values at birth. However, the organ remained more fatiguable than that of the adult perhaps due to limited transmitter synthesis by the sub-adult concentrations of choline acetyltransferase at the nerve terminals.

Distribution of putative amino acid transmitters, choline acetyltransferase and glutamate decarboxylase in the inferior colliculus

Levels of glutamic acid, aspartic acid, glycine, taurine, alanine, choline acetyltransferase, and glutamate decarboxylase were measured in ten regions of the inferior colliculus in the cat. None of these substances were uniformity distributed in the colliculus. The highest concentrations of asparate occurred in central regions, whereas those of glycine were in ventral central regions. The distributions of glutamic acid and glutamate decarboxylase were highly correlated, with both being generally high in dorsal regions. There was also a high correlation in the distributions of taurine and alanine, with these substances having their highest concentrations in the posterior region and lowest concentrations in the anterior region. The concentration of choline acetyltransferase was highest in the anterior region. No consistent changes in any of the substances were seen following elimination of inputs ascending to the colliculus via the dorsal and intermediate acoustic striae. Small decreases in glutamic acid were seen in two extracentral regions following removal of the auditory cortex. The nonuniform distribution of the substances that were analyzed suggest that there may be previously undescribed differences in the structure and function of the inferior colliculus. The lack of change in the content of choline acetyltransferase and glutamate decarboxylase after lesion of inputs from the cortex or cochlear nucleus makes it unlikely that acetylcholine or α-aminobutyrate are transmitters in these pathways.

Immunochemical demonstration of reversible reduction in choline acetyltransferase concentration in rat hypoglossal nucleus after hypoglossal nerve transection.

STUDIES of the cellular reaction of peripheral and central catecholaminergic neurones to axonal injury (the retrograde reaction and axon response) have demonstrated that such lesions elicit a prolonged reduction within the parent cell body in the activities and amounts of the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH) and, in noradrenergic neurones, dopamine-β-hydroxylase (DBH)1–3. The reduction in enzyme protein, usually reversible, lasts for several weeks and is apparently due to a decrease in the rate of enzyme biosynthesis4. It has therefore been proposed that a reduction in amount and activity of neurotransmitter biosynthetic enzymes may be a biochemical indicator of the retrograde reaction3. However, it remains to be established if a reversible diminution of neurotransmitter biosynthetic enzyme occurs during the retrograde reaction in neurones which synthesise neurotransmitters other than catecholamines. This is suggested from studies of the retrograde reaction in hypoglossal neurones, cholinergic motor neurones which innervate the tongue through the hypoglossal nerve. During the retrograde reaction elicited by lesions of the hypoglossal nerve, the activities of choline acetyhransferase (CAT), the enzyme synthesising acetylcholine, and acetyl-cholinesterase (AChE), the enzyme degrading it, are markedly reduced5–7. If the proximal stump of the lesioned nerve is allowed to regenerate and reestablish synaptic contact, AChE activity within the hypoglossal nucleus rapidly reappears7. It is, however, unknown whether the reduction in CAT activity is reversible and, because of a lack of specific, inhibiting antibody to CAT, whether the reduction is due to a decrease of enzyme molecules. We report here that the reduction in CAT in hypoglossal neurones elicited by transection of the hypoglossal nerve is reversible and also, by use of a specific antibody, that it is due to a decrease in the amount of CAT enzyme protein.

Alterations of acetylcholine enzymes in neuroblastoma cells persistently infected with lymphocytic choriomeningitis virus

AbstractPersistent infection of murine neuroblastoma cells with a relatively non‐cytopathic virus, lymphocytic choriomeningitis virus (LCMV), significantly lowered the cells' concentrations of choline acetyl transferase (CAT) and acetylcholine esterase (ACHE), enzymes which make or degrade acetylcholine. Quantities of acetylcholine enzymes remained depressed during the observation period of more than two years. This cellular luxury function was turned off without observable alterations in the cells' vital functions – growth rates, protein and RNA synthesis. Cloning experiments showed that CAT and ACHE levels were altered in the majority of LCMV infected neuroblastoma cells in culture and not limited to a specific subpopulation. Cells persistently infected with virus also contained receptors for neurotoxin A and α bungarotoxin. Six months after becoming infected, neuroblastoma cells having significant alterations in luxury functions stopped making infectious virus. Instead these cells now produced a defective interfering virus component.Similar events to those seen in vitro with neuroblastoma cells also occurred in vivo. Mice inoculated with LCMV at birth carried high titers of LCMV in brain tissues and viral antigens in neuronal cells as adults. Some of these mice also showed significant alterations in their ability to make and degrade acetylcholine when compared to age and sex matched controls.