Nucleus A10 dopaminergic neurons in inbred mouse strains: Firing rate and autoreceptor sensitivity are independent of the number of cells in the nucleus

Inbred mouse strains have different numbers of midbrain dopaminergic neurons; for example, BALB/cJ mice have 20–25% more neurons than CBA/J mice. As the number of cells decrease, for example in Parkinson's disease and in animals with midbrain dopaminergic cell lesions, the activity of their remaining cells increases. The purpose of the present experiment was to determine whether the functional properties of dopaminergic neurons in the ventral tegmental area (nucleus A10) differ in inbred mouse strains which possess different numbers of cells. The firing rate and autoreceptor sensitivity of A10 dopaminergic cells were examined in the in vitro slice preparation in BALB/cJ, C3H/HeJ, CBA/J, and DBA/2J mouse strains. It was observed that the autoreceptors on mouse dopaminergic neurons exhibit pharmacological properties of dopamine autoreceptors; activation of the autoreceptor produced a marked inhibition (50–70%) in cell firing rate by quinpirole (10⁻⁸ M), LY-141865 (10⁻⁷ M), (+)-3-(3-hydroxyphenyl)-N-n-propyl-piperidine (10⁻⁶ M), propyl-norapomorphine (10⁻⁵ M) and dopamine (10⁻⁴ M), and this inhibition was blocked or reversed by specific dopamine D2 receptor antagonists [(−) sulphide and spiroperidol, 10⁻⁶ M]. The baseline firing rates of the A10 cells did not differ among the four inbred strains [range 2.5 ± 0.2 (C3H/HeJ)−3.4 ± 0.3 (CBA/J) spikes/s ±SEM], and there was no significant difference in autoreceptor sensitivity among the mouse strains as assessed either by superfused dopamine (inhibitory dose 50% ≈150 μM), or by superfused quinpirole (inhibitory dose 50% ≈10 nM). These data indicate that differences in A10 cell numbers of 30% do not significantly influence the baseline firing rate or autoreceptor sensitivity of the cells.     

Phenytoin desensitization monitored by antigen specific T cell response using carboxyfluorescein succinimidyl ester dilution assay

We evaluated drug-specific T cell responses in a patient with refractory partial seizures and paroxysmal kinesigenic choreoathetosis successfully treated with clinical desensitization to phenytoin. Drug-induced lymphocyte transformation test before desensitization was negative with a stimulation index of 130%. The frequencies and cytokine-producing phenotypes of phenytoin-specific T cells were examined simultaneously by using a carboxyfluorescein succinimidyl ester (CFSE) dilution assay. Before desensitization, the proportion of CFSElow CD4+ cells in whole CD4+ was 3.09%; 13.6% of CFSElow CD4+ cells were stained with anti-interferon gamma antibody. After desensitization, phenytoin-specific CFSElow CD4+ cells decreased to background level. These results indicate that CFSE dilution assay will be useful for the diagnosis and monitoring of drug hypersensitivity.     

Long-lasting effect of ceruletide on dyskinesia and monoaminergic neuronal pathways in rats treated with iminodipropionitrile

In a model of dyskinesia induced by the administration of iminodipropionitrile (IDPN) in the rat, we evaluated the effects of ceruletide, an analogue of cholecystokinin, on behavioral abnormalities and monoaminergic neuronal function. Vertical head twitching in the IDPN-treated animals was inhibited for over 5 h following a single subcutaneous dose of 160 micrograms/kg ceruletide. In animals dosed daily for 2 or 3 days, the number of head twitches at 24 h after the last dose was about one-third of the number before treatment. After repeated daily doses of ceruletide for 6 days, the number of head twitches was reduced to low levels and remained significantly below pretreatment levels until the 4th posttreatment day. These results indicate that the inhibition of dyskinesia by ceruletide was long-lasting. Assays of monoaminergic neurotransmitters and their metabolites in various brain regions indicate that an imbalance between dopaminergic and serotonergic neuronal systems plays a major role in the pathogenesis of the IDPN-induced dyskinesia, i.e. the ratio of (DOPAC+HVA)/5-HIAA was significantly greater in the striatum but significantly smaller in the hippocampus of the IDPN-treated vs normal animals. This initially abnormal ratio of (DOPAC+HVA)/5-HIAA in the striatum and hippocampus of IDPN-treated animals returned to normal following treatment with ceruletide, corresponding with the reduction of the head twitching. The alterations in monoaminergic neuronal function induced by repeated administration of ceruletide persisted for at least 3 days, even though its plasma half-life is several minutes. Ceruletide also exerted a marked effect on monoaminergic neuronal function in the IDPN-treated rats, in contrast to only a slight effect in normal animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

An animal model of chronic inflammatory pain: pharmacological and temporal differentiation from acute models.

Clinically, inflammatory pain is far more persistent than that typically modelled pre-clinically, with the majority of animal models focussing on short-term effects of the inflammatory pain response. The large attrition rate of compounds in the clinic which show pre-clinical efficacy suggests the need for novel models of, or approaches to, chronic inflammatory pain if novel mechanisms are to make it to the market. A model in which a more chronic inflammatory hypersensitivity phenotype is profiled may allow for a more clinically predictive tool. The aims of these studies were to characterise and validate a chronic model of inflammatory pain. We have shown that injection of a large volume of adjuvant to the intra-articular space of the rat knee results in a prolonged inflammatory pain response, compared to the response in an acute adjuvant model. Additionally, this model also results in a hypersensitive state in the presence and absence of inflammation. A range of clinically effective analgesics demonstrate activity in this chronic model, including morphine (3mg/kg, t.i.d.), dexamethasone (1mg/kg, b.i.d.), ibuprofen (30mg/kg, t.i.d.), etoricoxib (5mg/kg, b.i.d.) and rofecoxib (0.3-10mg/kg, b.i.d.). A further aim was to exemplify the utility of this chronic model over the more acute intra-plantar adjuvant model using two novel therapeutic approaches; NR2B selective NMDA receptor antagonism and iNOS inhibition. Our data shows that different effects were observed with these therapies when comparing the acute model with the model of chronic inflammatory joint pain. These data suggest that the chronic model may be more relevant to identifying mechanisms for the treatment of chronic inflammatory pain states in the clinic.     

Nurr1基因转染人脐血间充质干细胞诱导分化为多巴胺能神经元的实验研究

目的探讨外源性Nurrl基因修饰的人脐血间充质干细胞在基因重组成纤维细胞生长因子-8(FGF8)和音猬因子(Shh)诱导下体外分化为多巴胺能神经元的情况。方法间充质干细胞被随机分为A组(对照组)、B组(Nurrl组)、C组(FGF8+Shh组)及D组(Nurrl+FGF8+Shh),采用脂质体法将pcDNA3.1-Ntrr1转染B、D组间充质干细胞,Western blot观察Nurr1基因表达情况,并在神经元条件培养液中进行增殖培养和诱导分化,间接免疫荧光染色法鉴定细胞性质,高效液相色谱法测定多巴胺含量。结果Western blot结果显示转染后Nurr1蛋白表达明显增高。A组和B组未发现酪氨酸羟化酶(TH)阳性细胞,而C组和D组TH阳性细胞分别为10．12％±2．65％及25．36％±3．13％,多巴胺含量分别为(43．6±2．1)nmol／L及(83．2±3．5)nmol／L,差异均有显著性意义(P< 0．01)。结论人脐血间充质干细胞在FGF8和Shh诱导下可以分化为多巴胺能神经元,外源性Nurr1基因修饰后,分化为多巴胺能神经元的数量增加。     

Generation and survival of midbrain dopaminergic neurons in weaver mice

Generation and survival of midbrain dopaminergic (DA) neurons were investigated using tyrosine hydroxylase (TH) immunocytochemistry combined with tritiated thymidine autoradiography at appropriate anatomical levels throughout the anteroposterior (A/P) axes of the substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA). The wild-type (+/+) and homozygous weaver (wv/wv) mice used here were the offspring of pregnant dams injected with the radioactive precursor when the mesencephalic neurons were being produced (gestational days 11-15). Data reveal that, at postnatal day 90, depletion of TH-stained cells in the wv/wv presented an A/P pattern of increasing severity and, therefore, the DA cells located in posterior parts of the SNc or the VTA appear to be more vulnerable than the settled anterior neurons. When the time of neuron origin is inferred for each level of these cell groups, it is found that the neurogenesis span is similar for both experimental groups, although significant deficits in the frequency of wv/wv late-generated neurons were observed in any level considered. On the other hand, it has been found that TH-positive neurons were settled along the extent of the SNc and the VTA following precise and differential neurogenetic gradients. Thus, the acute rostrocaudal increase in the proportion of late-generated neurons detected in both+/+DA-cell groups is disturbed in the weaver homozygotes due to the indicated A/P depletion.     

苯丙胺诱发小鼠激怒反应及其机制

苯丙胺15mg/kg ip能诱发小鼠激怒反应,使小鼠出现攻击和殴斗行为。强安定药、弱安定药和利血平有明显拮抗效果。镇静剂如巴比妥类、抗胆碱药及抗肾上腺素药均无拮抗作用。金刚胺、左旋多巴和阿扑吗啡均能增强苯丙胺的激怒效应。因苯丙胺激怒小鼠的方法简便易行,可作为筛选抗精神病药的动物模型。苯丙胺产生激怒作用的机制与增强多巴胺能神经的功能有关,推测可能是促进边缘系统多巴胺释放的结果。     

香烟烟对变态反应性疾病的影响

为探讨香烟烟对变态反应性疾病的影响，以香烟烟作为变应原，采用特异性体内皮肤试验测定了１６０６例变态反应性疾病患者对香烟烟的反应，并对香烟烟呈阳性反应与各种变态反应性疾病的关系以及与性别和年龄的关系进行了多元回归分析。结果表明，对香烟烟的皮试阳性的反应率为３４．６％（５５６例患者对香烟烟呈阳性反应），香烟烟的阳性反应与变应性疾病的发病有关，而与性别、年龄、病程、外周血白细胞计数和嗜酸性粒细胞计数相关性不大。对５８例香烟烟呈阳性反应的支气管哮喘患者，用乙酰甲胆碱做了气道激发试验，其结果为，激发后的最大肺活量（ＦＶＣ）、一秒钟用力呼气量（ＦＥＶ１）、最大呼气流速（ＰＥＦＲ）低于激发前。对２２３例香烟烟阳性反应者（其中支气管哮喘患者１８４例、变应性鼻炎８７例、过敏性皮肤病５２例），采用ＥＬＩＳＡ方法测定血清总ＩｇＥ，并与正常对照组比较，香烟反应的病人组血清总ＩｇＥ显著增高。由此可见，香烟烟可以引起变态反应性疾病。     

The role of dopamine in the facilitatory effect of angiotensin II on impaired learning in rats chronically treated with ethanol

Rats with impaired active avoidance induced by chronic (9 weeks) administration of ethanol were studied. Angiotensin II (ANG II) administered (ICV, 2.0 g) 12 h after the withdrawal of the alcohol not only neutralized the toxic effect of ethanol but also improved learning. When administered on the 5th day after ethanol withdrawal, the effect of ANG II was weaker. Tests of stereotypy and catalepsy were used to study the possible role of the dopaminergic system in this action of ANG II. It was shown that both chronic alcohol treatment and ANG II alone increased apomorphine (1 mg/kg) and amphetamine (7.5 mg/kg) stereotypy but the effects of ANG II were greater. ANG II did not change the stereotypy induced by amphetamine but increased the stereotypy induced by apomorphine in the group of animals chronically treated with alcohol. Haloperidol — induced catalepsy was reduced in these rats. ANG II alone intensified catalepsy and eliminated the effect of ethanol. Both ANG II and alcohol increased striatal dopamine (DA) concentration. This effect of ANG II was significantly greater in the animals chronically treated with alcohol. The above changes were not observed after the DA level had been reduced by alpha-methyl-p-tyrosine (250 mg/kg), nor were changes observed in the striatal DOPAC. The results suggest involvement of the central dopaminergic system in the effect of ANG II on the ethanol — induced impairment of acquisition of active avoidance but, however, the results of the biochemical determinations of DA turnover do not provide an explanation of these changes.