Cholecystokinin modulation of apomorphine- or amphetamine-induced stereotypy in rats: opposite effects.

Stereotyped behavior can be induced by the dopamine agonist apomorphine or by the releasing agent amphetamine. Cholecystokinin influence on dopamine-mediated behaviors has been extensively studied but a real controversy remains. Our purpose was to further characterize the dopamine-cholecystokinin interaction in apomorphine- and amphetamine-induced stereotyped behavior using sulphated cholecystokinin octapeptide (CCK8) and cholecystokinin tetrapeptide (CCK4) treatments. The results showed that CCK8 decreases apomorphine-induced stereotyped behavior and CCK4 has no effect. CCK4 and CCK8 increased the amphetamine-induced stereotyped behavior; CCK4 was more effective. The results confirm the opposite modulation of apomorphine or amphetamine-induced stereotyped behavior by CCK. These data suggest that this modulation is mediated by both CCK receptors on apomorphine-induced and only by CCK(2) receptors on amphetamine-induced stereotyped behavior.     

Behavioral investigation of the coexistence of substance P,corticotropin releasing factor,and acetylcholinesterase in lateral dorsal tegmental neurons projecting to the medial frontal cortex of the rat

Colocalization of substance P (SP), corticotropin releasing factor (CRF), and acetylcholinesterase (AChE) was detected by retrograde tracing and immunocytochemical staining in the nucleus tegmentalis dorsalis lateralis (ntdl) projecting to the medial frontal cortex (MFC), septum, and thalamus of the rat. The histochemical results suggest that SP and CRF coexist within a subpopulation of ntdl cholinergic neurons that project to a number of forebrain regions including the MFC. Behavioral studies of the effects of SP, CRF, and the cholinergic agonist, carbachol, employed microinjections into the MFC of rats. SP and CRF did not elicit any behavioral effects when administered alone. Carbachol (1–5 μg/side) produced a stereotyped motor behavior, consisting of rapid forepaw treading while in an upright posture, resembling “boxing.” SP (1 μg/side) increased carbachol-induced “boxing.” CRF (1–10 ng/side) decreased carbachol-induced “boxing.” One possible functional significance of the coexistence of SP, CRF, and acetylcholinesterase, in neurons projecting to the medial frontal cortex in rats, appears to be a modulatory potentiation of cholinergic response by SP, and a modulatory inhibition of the cholinergic response by CRF.     

Genetic identification of AChE as a positive modulator of addiction to the psychostimulant D-amphetamine in zebrafish

Addiction is a complex maladaptive behavior involving alterations in several neurotransmitter networks. In mammals, psychostimulants trigger elevated extracellular levels of dopamine, which can be modulated by central cholinergic transmission. Which elements of the cholinergic system might be targeted for drug addiction therapies remains unknown. The rewarding properties of drugs of abuse are central for the development of addictive behavior and are most commonly measured by means of the conditioned place preference (CPP) paradigm. We demonstrate here that adult zebrafish show robust CPP induced by the psychostimulant D-amphetamine. We further show that this behavior is dramatically reduced upon genetic impairment of acetylcholinesterase (AChE) function in ache/+ mutants, without involvement of concomitant defects in exploratory activity, learning, and visual performance. Our observations demonstrate that the cholinergic system modulates drug-induced reward in zebrafish, and identify genetically AChE as a promising target for systemic therapies against addiction to psychostimulants. More generally, they validate the zebrafish model to study the effect of developmental mutations on the molecular neurobiology of addiction in vertebrates.     

Integrative gene network analysis provides novel regulatory relationships, genetic contributions and susceptible targets in autism spectrum disorders

Autism spectrum disorders (ASDs) are a group of diseases exhibiting impairment in social drive, communication/language skills and stereotyped behaviors. Though an increased number of candidate genes and molecular interactions have been identified by various approaches, the pathogenesis remains elusive. Based on clinical observations, data from accessible GWAS and expression datasets we identified ASDs gene candidates. Integrative gene network and a novel CNV-centric Node Network (CNN) analysis method highlighted ASDs-associated key elements and biological processes. Functional analysis identified neurological functions including synaptic cholinergic receptor (CHRNA) families, dopamine receptor (DRD2), and correlations between social behavior and oxytocin related pathways. CNN analysis of genome-wide genetic and expression data identified inheritance-related clusters related to PTEN/TSC1/FMR1 and mTor/PI3K regulation. Integrative analysis identified potential regulators of networks, specifically TNF and beta-estradiol, suggesting a potential central role in ASDs. Our data provide information on potential disease mechanisms, and key regulators that may generate novel postulations, and diagnostic molecular biomarkers.     

圈养马麝刻板行为表达与粪样类固醇激素的关系

2017年7月1日-8月31日及2018年6月1日-7月31日,在甘肃兴隆山保护区马麝繁育中心,采用焦点取样法和连续记录法进行了圈养马麝的刻板行为取样,采集同期粪样,并用放射免疫分析法（RIA）检测粪样中肾上腺皮质醇、睾酮及雌二醇激素的水平,分析了圈养马麝刻板行为表达与上述3种激素水平的关系。结果显示,展现刻板行为的圈养马麝的皮质醇水平（111.099 ± 16.231）ng/g略高于无刻板行为表达的马麝（95.640± 9.738） ng/g,差异未达显著（P> 0.05）;展现刻板行为雄麝的睾酮水平（135.900± 21.582）ng/g略高于无刻板行为的雄麝（108.182 ± 9.689） ng/g,差异也不显著（P> 0.05）;展现刻板行为雌麝的雌二醇水平（0.445 ± 0.116）ng/g显著低于无刻板行为雌麝（10.843 ± 1.142）ng/g（P< 0.05）。研究结果表明,圈养雄性马麝的刻板行为表达与其类固醇激素水平不相关;而雌麝的刻板行为表达与雌二醇分泌显著负相关,这与其繁殖及健康状况有关。在麝类驯养实践中,可将粪样类固醇激素水平（尤其是雌二醇）作为其受胁迫水平及行为健康的监测指标。     

Genetic identification of AChE as a positive modulator of addiction to the psychostimulant D‐amphetamine in zebrafish

Addiction is a complex maladaptive behavior involving alterations in several neurotransmitter networks. In mammals, psychostimulants trigger elevated extracellular levels of dopamine, which can be modulated by central cholinergic transmission. Which elements of the cholinergic system might be targeted for drug addiction therapies remains unknown. The rewarding properties of drugs of abuse are central for the development of addictive behavior and are most commonly measured by means of the conditioned place preference (CPP) paradigm. We demonstrate here that adult zebrafish show robust CPP induced by the psychostimulant D‐amphetamine. We further show that this behavior is dramatically reduced upon genetic impairment of acetylcholinesterase (AChE) function in ache/+ mutants, without involvement of concomitant defects in exploratory activity, learning, and visual performance. Our observations demonstrate that the cholinergic system modulates drug‐induced reward in zebrafish, and identify genetically AChE as a promising target for systemic therapies against addiction to psychostimulants. More generally, they validate the zebrafish model to study the effect of developmental mutations on the molecular neurobiology of addiction in vertebrates. © 2006 Wiley Periodicals, Inc. J Neurobiol, 2006     

The effect of housing and environmental enrichment on stereotyped behavior of adult vervet monkeys (Chlorocebus aethiops)

Little information is available on the response of vervet monkeys to different housing conditions or on the suitability of enrichment devices or methods for vervet monkeys. In this study, the authors evaluated the occurrence of stereotyped behavior in adult vervet monkeys under various conditions of housing and enrichment. The variables included cage size, cage level (upper or lower), enrichment with a foraging log, enrichment with an exercise cage and presence of a mate. The authors first determined the incidence of stereotyped behavior in captive-bred, singly housed adult female and male vervet monkeys. They then exposed monkeys to different housing and enrichment situations and compared the incidence of stereotyped behavior among the monkeys. The authors found that more females than males engaged in stereotyped behavior and that females, on average, engaged in such behavior for longer periods of time than males. Stereotyped behavior was most often associated with a small, single cage. The average amount of observed stereotyped activity in monkeys housed in a small cage was significantly lower when the monkeys had access to either a foraging log or an exercise cage. Stereotyped behavior was also lower in female monkeys that were housed (either with a male or without a male) in a larger cage. The least amount of abnormal behavior was associated with the largest, most complex and enriched housing situation. Males and females housed in cages on the lower level of two-level housing engaged in more stereotyped behavior than did monkeys housed in the upper level, regardless of the presence or type of enrichment provided.     

Phencyclidine-induced stereotyped behavior in rats: dose response effects and antagonism by neuroleptics.

Phencyclidine hydrochloride produced a very characteristic and reproducible stereotyped behavioral syndrome in rats. Both the intensity and the duration of the phencyclidine-induced stereotyped behavior are elicited in a dose-dependent manner in the 2–16 mg/kg dose range. The predominant behavior elicited by low doses was repetitive lateral head swaying, while with higher doses circling and backward walking were observed in addition to the head swaying. This behavior was antagonized by the neuroleptic agents chlorpromazine, haloperidol, and pimozide, but not by α- or β-adrenergic blockers. These results indicate that the phencyclidine-induced stereotyped behavior may be mediated by central dopaminergic mechanisms.     

Cholinergic induction of seizures in the rat prefrontal cortex

Intracerebral microinjection of the cholinergic agonist, carbachol, into the medial prefrontal cortex of the rat, induced a profound behavioral syndrome consisting of repetitive, stereotyped forepaw treading in an upright posture. Electroencephalographic analysis revealed multiple bursts of sharp waves, 200-300 microV, accompanying the carbachol-elicited motor behavior. Pretreatment with intraperitoneal doses of three anticonvulsant drugs, clonazepam, diazepam, and pentobarbital, blocked the manifestation of the motor behavior. These observations suggest that activation of cholinergically innervated regions of the rat medial prefrontal cortex induces an atypical form of seizures.     

The influence of caffeine on d-amphetamine- and apomorphine-induced stereotyped behavior

The influence of caffeine on amphetamine- and apomorphine-induced stereotyped behavior in guinea pigs has been investigated. Caffeine potentiated amphetamine- and apomorphine-induced stereotyped behavior. These observations support the concept that a dopamine sensitive adenyl cyclase and cAMP play a role in mediating the effect of dopamine at specific striatal dopamine receptors.     

Estimation of deanol and choline by gas chromatography mass spectrometry

Analytical methods are described which permit the measurement of both deanol and choline in the same sample by gas chromatography mass spectrometry when either compound may be present in large excess (100:1). Deuterium labelling is employed for internal standards, to distinguish endogenous from tracer variants and to distinguish deanol in the sample from deanol formed by derivatization of choline. The limit of detection of both compounds is about 50 pmol.     

Pupariation in flies: A tool for monitoring effects of drugs,venoms, and other neurotoxic compounds

A complex Sarcophaga bullata pupariation assay was used to evaluate the neurotropic effects of several drugs, venoms, and insecticides. The assay consists of tests for (1) immediate effects on the intact larva, (2) effects on ligated (ie, isolated from the central nervous system) larval abdomens, (3) morphogenetic effects on the puparium, and (4) effects on stereotyped pupariation behavior. The latter are monitored barographically by recording changes in hemocoelic pressure. Of 62 compounds screened, 18 showed morphogenetic activity at a threshold dose of 5 μg or less, 11 at a dose of 50 μg, four at a dose of 100 μg, and 29 showed no morphogenetic activity. From a comparison of the putative pharmacological actions of the tested compounds with their morphogenetic effects, certain generalizations can be made: Agents that paralyze neuromuscular systems at the peripheral level (eg, tetrodotoxin), or suppress or modify basic motor patterns centrally (eg, veratrine sulphate), cause retention of larval morphological characters in the puparium. Compounds that stimulate convulsive contractions of segmental musculature (mostly cholinergic drugs like eserine sulphate, nicotine, organophosphate insecticides) cause retention of larval segmentation on longitudinally contracted puparia. Five compounds (venom of the scorpion, Leirus quinquestriatus, pyrethrins, protoveratrine A, and kainic and quisqualic acids) stimulate musculature of the denervated abdomen. Barographic monitoring of changes in pupariation behavior appears to be a most sensitive and informative test. It reveals great differences in the ways in which compounds producing seemingly identical morphogenetic effects affect and modify behavior, thus making pharmacological classification more accurate.     

The bombesin/gastrin releasing peptide receptor antagonist RC-3095 blocks apomorphine but not MK-801-induced stereotypy in mice

Bombesin (BN)-like peptides might be involved in the pathogenesis of neuropsychiatric disorders such as schizophrenia. Stereotyped behaviors induced by the dopamine receptor agonist apomorphine or the N-methyl-D-aspartate glutamate receptor antagonist dizocilpine (MK-801) in rodents have been proposed as animal models of schizophrenic psychosis. In the present study we evaluated the effects of the BN/gastrin-releasing peptide receptor (GRP) antagonist (D-Tpi6, Leu13 psi[CH2NH]-Leu14) bombesin (6-14) (RC-3095) on apomorphine and MK-801-induced stereotyped behavior in mice. An intraperitoneal (i.p.) injection of RC-3095 (1.0, 10.0 or 100.0 mg/kg) blocked apomorphine-induced stereotypy. The inhibitory effect of RC-3095 on apomorhine-induced stereotypy was similar to that induced by haloperidol (0.5 mg/kg). RC-3095 did not affect stereotyped behavior induced by MK-801 (0.5 mg/kg). The results provide the first evidence that BN/GRP receptor antagonism blocks stereotyped behavior induced by a dopamine agonist. Together with previous evidence, the present study indicates that the BN/GRP receptor can be considered a drug target in the investigation of potential new agents for treating neuropsychiatric disorders.     

Effect of feeding boxes on the behavior of stereotyping amur tigers (Panthera tigris altaica) in the Zurich Zoo,Zurich, Switzerland

The stereotyped pacing shown by the two Amur tigers in the Zurich Zoo was hypothesized as being caused by permanently frustrated appetitive foraging behavior. Several electrically controlled feeding boxes were installed and access to each box was possible only twice a day for 15 min at semi‐random times. The boxes had to be opened actively by the tigers. Two trials were carried out: one with solitary confinement, and one with paired confinement. During box feeding, the female's stereotyped pacing was significantly reduced from 16% (solitary confinement, conventional feeding) and 7% (paired confinement, conventional feeding) to 1% (solitary confinement) and less than 0.01% (paired confinement) of the daily observed time. The female's sleeping increased significantly in both solitary and paired confinement. The male only showed a significant reduction in stereotyped pacing behavior when kept with the female (conventional feeding: 10%; box feeding: <0.01% of the daily observed time). On days with a box‐feeding regime in paired confinement, the male spent 25% (83 min) of the observed time with active behavior at the feeding boxes. The results support the hypothesis that permanently frustrated appetitive foraging behavior causes stereotyped pacing in adult tigers. Zoo Biol 21:573–584, 2002. © 2002 Wiley‐Liss, Inc.     

Muscarinic cholinergic receptor blockade impairs free fatty acid mobilization during fasting in pigeons (<Emphasis Type="Italic">Columba livia</Emphasis>)

The effects of intracerebroventricular pretreatment with muscarinic (scopolamine or methylscopolamine; 2.7 nmol or 5.4 nmol) or nicotinic (mecamylamine, 2.7 nmol or 5.4 nmol) cholinergic receptor antagonists on plasma free fatty acid increases induced by intracerebroventricular injections of carbachol in conscious resting pigeons (Columba livia) were examined. Plasma glucose levels were also measured throughout the experiments. Pretreatment with methylscopolamine suppressed the lipolytic effect of carbachol injections, while mecamylamine left this response unchanged. Neither carbachol treatment alone, nor the pretreatments with cholinergic agents affected glucose levels. Subsequently, the effects of intracerebroventricular injections of methylscopolamine were investigated in 24-h food-deprived pigeons. The increase in free fatty acid levels after fasting was of a magnitude similar to that observed after carbachol treatment; intracerebroventricular injections of methylscopolamine (5.4 nmol) transiently but powerfully decreased plasma free fatty acids in 24-h food-deprived pigeons to levels comparable to those of free-feeding animals. The fasting-induced decrease in glucose levels was not affected by this treatment. These data indicate that the lipolytic response induced by carbachol may be mediated by central muscarinic cholinergic receptors and that this central cholinergic mechanism partially contributes to plasma free fatty acid increases observed during fasting. Furthermore, the absence of effects on glucose levels suggests that these cholinergic mechanisms participate selectively in the lipolytic component of the metabolic response to fasting.     

Evidence of serotonin-dopamine involvement in the inhibition of d-amphetamine stereotypy and appearance of stereotyped movements found respectively after acute and long-term treatment with morphine and methadone in rats

P-chlorophenylalanine, an inhibitor of serotonin synthesis, was found to completely prevent the inhibitory effect of morphine and methadone on the stereotypy caused by d-amphetamine and methyl-phenidate in rats. d-Fenfluramine and m-chlorophenylpiperazine, two drugs supposed to increase serotonin transmission, and halo-peridol, an antagonist of dopamine at central receptors, blocked the stereotyped movements induced by repeated treatment with morphine and methadone. The results suggest that a) brain serotonin mediates the effect of morphine and methadone on amphetamine and methylphenidate stereotypy b) serotonin and dopamine are involved in the stereotyped movements caused by long-term treatment with these narcotics in the rat.     

Centrally administered vasopressin antagonizes pentobarbital-induced narcosis and depression of hippocampal cholinergic activity

Intracerebroventricular (ICV) microinjection of arginine vasopressin (AVP) to pentobarbital-anesthetized rats produced shortening of the duration of narcosis. This analeptic effect was blocked by atropine, indicating the central cholinergic nature of the response. AVP also increased hippocampal sodium-dependent high affinity choline uptake activity that had been depressed by the barbiturate. The AVP analeptic effect was blocked by pretreatment with a V-1 (vasopressor), but not a V-2 (antidiuretic), vasopressin receptor antagonist. These results suggest that ICV AVP produces its analeptic effect by interacting with central V-1 receptors to activate a hippocampal cholinergic arousal system. The cholinergic arousal effect may be a factor in the memory enhancing property of AVP.     

Osmoprotective activity, urea protection, and accumulation of hydrophilic betaines in Escherichia coli and Staphylococcus aureus

The hydrophilic betaines, deanol betaine, triethanol betaine, diethanolthetin and methylethanolthetin, and also thioxanium betaine and citrulline betaine, were accumulated by Escherichia coli. All betaines tested had significant osmoprotective activity for E. coli and, with the exception of citrulline betaine and diethanolthetin, also demonstrated urea protection. Staphylococcus aureus accumulated only methylethanolthetin, deanol betaine and thioxanium betaine: the first two had an osmoprotective effect but conferred no urea protection. Diethanolthetin and thioxanium betaine significantly decreased urea tolerance for S. aureus.     

Effect of amizil and the ACTH4-10 fragment on the ability of mice to extrapolate the direction of movement

Mice possessing the ability to extrapolate the direction of movement (100% of correct choices) were injected i.p. with different doses of m-cholinolytic amizil 2 hours before experiment. Doses of 2-5 mg/kg reduced the percentage of correct choices, the adequate solving strategy being replaced by stereotyped unidirectional reactions or stereotyped alternating responses. Doses of 8-12 mg/kg induced "refusals" to solve extrapolation problem. When amizil treated mice were intraventricularly injected with 1 mg/kg of ACTH4-10, their extrapolation ability was restored. This compensatory action of peptide could be mediated by its influence on cholinergic as well as on other neurotransmitter systems.     

Octreotide-induced drinking, vasopressin, and pressure responses: role of central angiotensin and ACh

The involvement of central angiotensinergic and cholinergic mechanisms in the effects of the intracerebroventricularly injected somatostatin analog octreotide (Oct) on drinking, blood pressure, and vasopressin secretion in the rat was investigated. Intracerebroventricular Oct elicited prompt drinking lasting for 10 min. Water consumption depended on the dose of Oct (0.01, 0.1, and 0. 4 microgram). The drinking response to Oct was inhibited by pretreatments with the intracerebroventricularly injected angiotensin-converting enzyme inhibitor captopril, the AT(1)/AT(2) angiotensin receptor antagonist saralasin, the selective AT(1) receptor antagonist losartan, or the muscarinic cholinergic receptor antagonist atropine. The dipsogenic effect of Oct was not altered by prior subcutaneous injection of naloxone. Oct stimulated vasopressin secretion and enhanced blood pressure. These responses were also blocked by pretreatments with captopril or atropine. Previous reports indicate that the central angiotensinergic and cholinergic mechanisms stimulate drinking and vasopressin secretion independently. We suggest that somatostatin acting on sst2 or sst5 receptors modulates central angiotensinergic and cholinergic mechanisms involved in the regulation of fluid balance.