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作者： Cheung, WMW Fu, WY Hui, WS Ip, NY Hong Kong Univ Sci & Technol Dept Biol Clear Water Bay Hong Kong Peoples R China Hong Kong Univ Sci & Technol Biotechnol Res Inst Hong Kong Peoples R China

When treated with retinoic acid (RA), a human embryonal carcinoma (EC) cell line, NTera2 cl.D/1 (NT2), differentiates into several morphologically distinct cell types, which include terminally differentiated postmitotic central nervous system (CNS) neurons. Accumulating evidence has demonstrated the significant potential of NT2 cells in studies related to cancer therapy and neurodegenerative diseases However, preparation of enriched NT2 neurons often requires a lengthy period (co. five,weeks) and depends largely on tedious techniques similar to those used for primary neuronal cultures. Here, we report a rapid protocol for the preparation of these human CNS neurons. Using the method of cell aggregation, enriched NT2 neurons can he obtained in approximately two weeks. We also demonstrated that cell aggregation reduced the time normally required for the induction of neuronal differentiation as revealed by the early expression of neuronal markers. The period of RA treatment could also be reduced if NT2 cells were maintained cls aggregates for a sufficient period of time. Taken together our findings demonstrated that cell aggregation promoted RA-induced neuronal differentiation of NT2 cells and provided a rapid protocol for the efficient production of NT2 neurons. The ability to produce large quantities of human CNS neurons should facilitate future rise of these neurons for basic research and applications in cell therapy.

关键词：生物技术癌胚胎性/病理学细胞集合细胞分化/药物作用中枢神经系统/细胞学中枢神经系统/药物作用中枢神经系统/代谢神经组织蛋白质类/代谢神经突/超微结构神经元/细胞学神经元/药物作用神经元/代谢时间因素维甲酸/药理学肿瘤细胞培养的人类

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Role of high-affinity receptors and membrane transporters in nonsynaptic communication and drug action in the central nervous system

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PHARMACOLOGICAL REVIEWS 2000年第1期52卷 63-89页

作者： Vizi, ES Hungarian Acad Sci Inst Expt Med Dept Pharmacol H-1450 Budapest Hungary

Neurochemical and morphological evidence has shown that some neurotransmitters or substances may be released from both synaptic and nonsynaptic sites for diffusion to target cells more distant than those observed in regular synaptic transmission. There are functional interactions between neurons without synaptic contacts, and matches between release sites and localization of receptors sensitive to the chemical signal are exceptions rather than the rule in the central nervous system. This also indicates that besides cabled information signaling (through synapses), there is a "wireless" nonsynaptic interaction between axon terminals. This would be a form of communication transitional between discrete classical neurotransmission (in Sherrington's synapse) and the relatively nonspecific neuroendocrine secretion. Recent findings indicate that in addition to monoamines (norepinephrine, dopamine, serotonin), other transmitters, such as acetylcholine and nitric oxide (NO), may also be involved in these nonsynaptic interactions. It has been shown that NO, an ideal mediator of nonsynaptic communication, can influence the function of uptake carrier systems, which may be an important factor in the regulation of extracellular concentration of different transmitters. This review will focus on the role of nonsynaptic receptors and transporters in presynaptic modulation of chemical transmission in the central nervous system. The nonsynaptic interaction between neurons mediated via receptors and transports of high affinity not localized in synapses has the potential to be an important contributor to the properties and function of neuronal networks. In addition, it will be suggested for the first time that the receptors and transporters expressed nonsynaptically and being of high affinity are the target of drugs taken by the patient.

关键词：细胞间通讯/药物作用细胞间通讯/生理学中枢神经系统/药物作用中枢神经系统/生理学膜/药物作用膜/代谢受体神经递质/药物作用受体神经递质/生理学突触传递/药物作用突触传递/生理学动物人类

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Central autonomic activation by intracisternal TRH analogue excites gastric splanchnic afferent neurons

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JOURNAL OF NEUROPHYSIOLOGY 1999年第2期81卷 682-691页

作者： Adelson, DW Wei, JY Yashar, M O-Lee, TJ Cure, YT W Los Angeles Vet Affairs Med Ctr CURE Digest Dis Res Ctr Div Digest Dis Dept Med Los Angeles CA 90073 USA Univ Calif Los Angeles Inst Brain Res Los Angeles CA 90073 USA

Intracisternal (ic) injection of thyrotropin-releasing hormone (TRH) or its stable analogue RX 77368 influences gastric function via stimulation of vagal muscarinic pathways. In rats, the increase in gastric mucosal blood flow evoked by a low ic dose of RX 77368 occurs via release of calcitonin gene-related peptide from capsaicin-sensitive afferent neurons, most probably of spinal origin. In this study, the effect of low ic doses of RX 77368 on afferent impulse activity in splanchnic single fibers was investigated. The cisterna magna of overnight-fasted, urethan-anesthetized Sprague-Dawley rats was acutely cannulated, and fine splanchnic nerve twigs containing at least one fiber responsive to mechanical probing of the stomach were isolated at a site immediately distal to the left suprarenal ganglion. Unit mechanoreceptive fields were encountered in all portions of the stomach, both superficially and in deeper layers. Splanchnic afferent unit impulse activity was recorded continuously during basal conditions and in response to consecutive ic;injections of saline and RX 77368 (15-30 min later;1.5 or 3 ng). Basal discharge rates ranged from 0 to 154 impulses/min (median = 10.2 impulses/min). A majority of splanchnic single units with ongoing activity increased their mean discharge rate by greater than or equal to 20% after ic injection of RX 77368 at either 1.5 ng (6/10 units;median increase 63%) or 3 ng (19/24 units;median increase 175%). Five units lacking impulse activity in the 5-min before ic RX 77368 (3 ng) were also excited, with the onset of discharge occurring within 1.0-5.0 min postinjection. In units excited by ic RX 77368, peak discharge occurred 15.6 +/- 1.3 min after injection and was followed by a decline to stable activity levels less than or equal to 20-40 min thereafter. Tn a few cases (4/24), ic RX 77368(3 ng) inhibited the impulse activity of initially active units, with a time course comparable to that seen in units excited by the same treatment. T

关键词：自主神经系统/药物作用自主神经系统/生理学导管插入术中枢神经系统/药物作用中枢神经系统/生理学诱发电位/药物作用诱发电位/生理学注射脑室内神经元传入/药物作用神经元传入/生理学物理刺激吡咯烷酮羧酸/类似物和衍生物大鼠 Sprague-Dawley 内脏神经/药物作用内脏神经/生理学胃/神经支配胃/生理学促甲状腺素释放激素/类似物和衍生物促甲状腺素释放激素/药理学时间因素动物男(雄)性大鼠

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Nicotine infusion modulates immobilization stress-triggered induction of gene expression of rat catecholamine biosynthetic enzymes

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第2期291卷 884-892页

作者： Serova, L Danailov, E Chamas, F Sabban, EL New York Med Coll Dept Biochem & Mol Biol Valhalla NY 10595 USA

The relationship between nicotine and stress is complex and paradoxical. Although people claim they smoke because it relaxes them, nicotine can trigger some of the effects observed with stress, including the release and synthesis of the catecholamines and their biosynthetic enzymes. This study examined one aspect of this confusing relationship between nicotine and stress. Multiple injections of nicotine bitartrate (5 mg/kg) elevated mRNA levels for the catecholamine biosynthetic enzymes, tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine N-methyltransferase, and of preproneuropeptide Y in rat adrenal medulla more than did 1 mg/kg of nicotine bitartrate. In the locus ceruleus, substantia nigra, and ventral tegmental area both doses equally induced TH mRNA levels. Nicotine infusion (15 mg/kg/day) did not affect adrenal mRNA levels for any of the genes of interest and did not increase plasma corticosterone levels. However, in rats preexposed to nicotinic infusions, the response to a single immobilization (IMO) stress was markedly attenuated with respect to changes in adrenomedullary TH, DBH, and phenylethanolamine N-methyltransferase mRNA levels and in c-Fos protein levels. In the central nervous system, the chronic infusion of nicotine prevented the induction of TH mRNA by repeated IMO stress in the ventral tegmental area (but not in substantia nigra) and of DBH mRNA by single IMO in the locus ceruleus. These findings may explain some of the complex interactions between stress and exposure to nicotine.

关键词：肾上腺髓质/药物作用肾上腺髓质/酶学儿茶酚胺类/生物合成中枢神经系统/药物作用中枢神经系统/酶学皮质酮/血液多巴胺β羟化酶/遗传学混合功能氧化酶类/遗传学神经肽类/遗传学尼古丁/投药和剂量尼古丁/药理学苯基乙醇胺N-甲基转移酶/遗传学原癌基因蛋白质c-fos/代谢 RNA 信使/代谢大鼠 Sprague-Dawley 无特异性病原体有机物应力物理酪氨酸单氧化酶/遗传学动物男(雄)性大鼠

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ELECTROENCEPHALOGRAPHIC AND BEHAVIORAL-CHANGES ASSOCIATED WITH PAPAVERINE ADMINISTRATION IN HEALTHY GERIATRIC SUBJECTS

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JOURNAL OF THE AMERICAN GERIATRICS SOCIETY 1975年第7期23卷 295-300页

作者： COLE, JO BRANCONNIER, RJ MARTIN, GF BOSTON STATE HOSP INST RES & REHABILBOSTONMA 02124 BOSTON STATE HOSP GERIATRIC PSYCHOPHARMACOL GRPBOSTONMA 02124 SALEM HOSP MENTAL HLTH CTR OUTPATIENT DEPTSALEMMA

A double‐blind cross‐over procedure was used to evaluate the effects of papaverine hydrochloride on the electroencephalogram (EEG), cognition, mood and psychologic test performance of 10 healthy geriatric volunteers. During the two‐week period, each subject was tested four times: before and after receiving papaverine and before and after receiving placebo. Period analysis revealed that a 300‐mg dose of papaverine (Pavabid Plateau Caps) increased EEG alpha activity (8–13 cycles per second) and decreased the beta2 range (27–40 cps). Baseline values for six mood factors, short‐term memory, complex problem solving, and attention were not altered significantly by either drug or placebo. Results of the Subject Paced Digit Symbol Substitution Test indicated that papaverine may produce some improvement in simple cognitive functioning. No adverse effects were associated with the use of papaverine.

关键词： α节律行为/药物作用 β节律中枢神经系统/药物作用临床试验(主题) 认知/药物作用脑电描记术情绪/药物作用罂粟碱/药理学安慰剂心理学试验韦氏智力量表女(雌)性人类男(雄)性中年人

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Pharmacological regulation of the cough reflex - from experimental models to antitussive effects in man

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PULMONARY PHARMACOLOGY & THERAPEUTICS 1999年第4期12卷 215-228页

作者： Karlsson, JA Fuller, RW Rhone Poulenc Rorer Discovery Biol Dagenham England Glaxo Wellcome Inc Respiratory Int Med Affairs Uxbridge Middx England

A wide range of pharmacological agents have been examined in experimentally-induced cough in animal models and healthy human subjects and, to a lesser extent, in patients with chronic cough. Opioids, acting at a putative cough centre in the CNS, and inhaled local anaesthetics, suppressing afferent neural activity, remain the most effective antitussive drugs available in healthy subjects and in patients. Antiasthma drugs like β-agonists, theophylline and glucocorticoids suppress coughing in patients with asthma, but have little direct effect on experimental cough in healthy subjects or in subjects with chronic, dry cough. These data suggest that antiasthma drugs act indirectly by reducing inflammatory mediators in the bronchial mucosa that trigger coughing, rather than directly on any of the physiological components of the cough reflex. The use of cell cultures (e.g. electrophysiologica1 studies of nerve cell excitation), in vitro experiments and animal models are all essential for advancing our understanding of the physiology and pharmacology of respiratory reflexes, even though the ultimate confirmation of pathophysiological mechanisms require carefully conducted clinical experimental studies. Citric acid/capsaicin-induced coughing in conscious guinea-pigs is a most useful model of the cough reflex but it is clear that the antitussive activity of pharmacological agents is most predictive when the drugs are administered systemically. This particular model seems to have a propensity for identifying 'false-positive' drug candidates when they are administered locally to the airways. Further studies are required to refine the model and characterize it relative to pathological coughing in human subjects. There is no doubt that the drugs currently available for suppression of the cough reflex, most of which are sold OTC as part of various self-medication programmes, are inadequate and that there is an unmet medical need, particularly in patients with chronic, non-producti

关键词：镇咳药/药理学中枢神经系统/药物作用咳嗽/药物疗法咳嗽/病理生理学疾病模型动物反射/生理学动物豚鼠人类

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EFFECTS OF CYCLOPHOSPHAMIDE TREATMENT OF NEWBORN MICE ON DEVELOPMENT OF SWIMMING AND REFLEX BEHAVIOR AND ON ADULT BEHAVIORAL PERFORMANCE

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DEVELOPMENTAL PSYCHOBIOLOGY 1976年第6期9卷 555-567页

作者： PREACHE, MM GIBSON, JE MICHIGAN STATE UNIV DEPT PHARMACOL E LANSING MI 48824 USA

Cyclophosphamide (CP), an antineoplastic agent, was administered s.c. to Swiss-Webster mice on the day of birth and the mice were later tested for developmental or adult behavioral abnormalities. The CP dosages of 20, 30 or 45 mg/kg of body weight retarded maturation of swimming ability and 45 mg/kg retarded maturation of the righting reflex. At 7 wk of age mice treated neonatally with 30 or 45 mg/kg of CP had reduced locomotor activity and were more emotionally reactive than controls in an open field. Mice treated with 30 but not 20 mg/kg of CP tended to avoid shock less often than controls and those treated with 20 mg/kg fell more frequently when crossing a rotating rod for food. Rotorod performance was improved by treatment with 45 but not 30 mg/kg of CP. All dosages examined decreased body weight gains but only 30 or 45 mg/kg resulted in gross body malformations. The results indicate that CP can functionally impair the development of mice and that some of these impairments are independent of gross body malformations.

关键词：畸形药物性/病因学年龄因素动物新生/生长和发育觉醒/药物作用回避学习/药物作用行为动物/药物作用体重/药物作用中枢神经系统/药物作用环磷酰胺/药理学 DNA/生物合成剂量效应关系药物死亡率运动活动/药物作用运动技能/药物作用 RNA/生物合成反射/药物作用动物女(雌)性男(雄)性小鼠

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