尼古丁处理对大鼠脑内多巴胺转运体和酪氨酸羟化酶的影响

目的 观察尼古丁处理大鼠脑内多巴胺转运体(DAT)和酪氨酸羟化酶(TH)的表达变化,探讨尼古丁处理对大鼠脑内多巴胺(DA)能神经体系的影响. 方法 选用雄性Wistar大鼠按每日 0.4 mg/kg 腹腔注射尼古丁 7d;利用免疫组织化学和免疫印迹法,检测尼古丁处理大鼠有关脑区DAT和TH的表达改变. 结果 与对照组相比:1. 免疫组织化学显示,尼古丁处理组大鼠伏核(NACC)和腹则被盖区(VTA)的DAT灰度值降低了12.43 %和12.85 %;TH的灰度值则降低了11.87 %和10.09 %.2. 免疫印迹法显示,尼古丁处理组大鼠尾壳核(CPu)-NACC、黑质(SN)-VTA的DAT与β-肌动蛋白(β-actin)条带相对吸光度比值增加了75.68 %和117.14 %;而TH的比值则分别增加了66.32 %和60.31 %. 结论 尼古丁处理增加大鼠脑内DAT和TH的表达,这可能与尼古丁的成瘾机制有关.     

丙酸睾丸酮早期处理对大鼠行为及脑内多巴胺转运体的影响

目的:观察丙酸睾丸酮早期处理对大鼠行为及脑内多巴胺转运体的影响.方法:分别以开放广场实验及免疫组织化学技术观察丙酸睾丸酮处理大鼠行为及脑内多巴胺转运体(DAT)的变化.结果:开放广场实验显示早期注射丙酸睾丸酮可使大鼠活动增多,表现为水平运动、垂直运动、运动的总径长以及清洁动作次数显著增加;免疫组织化学显色结果表明丙酸睾丸酮处理组大鼠黑质和尾壳核DAT免疫反应强度增强,其灰度值显著低于对照组.结论:丙酸睾丸酮早期处理大鼠活动的增加可能与雄激素影响黑质-尾壳核多巴胺能神经体系有关.     

脑震荡鼠大脑多巴胺转运体变化研究

为了进一步探讨脑震荡(cerebral concussion,CC)大鼠认知行为障碍的神经生物学基础,本实验观察了CC大鼠多巴胺转运体(dopamine transporters,DAT)的变化规律。用自制金属单摆式闭合性脑损伤打击装置复制脑震荡大鼠模型48只,随机分为伤后1、2、4、8、16、24d六个实验组,另设一正常对照组(n=8)。运用抗DAT抗体SP法行免疫组化染色,并测定大脑前额叶皮质(PFC)、尾壳核(CPu)、伏隔核(Acbc)、内侧隔核(MS)、斜角带垂直臂核(VDB)、斜角带水平臂核(HDB)、梨形皮质(Pir)、顶叶皮质(CPL)、海马CA1-3、丘脑中央背内侧核(MD)、丘脑背外侧核(LD)、下丘脑腹内侧核(VMH)和杏仁核等16个脑区DAT阳性表达物的光密度值。结果表明,与正常对照组相比脑震荡后大鼠多个脑区DAT阳性表达物呈下调变化。PFC、CPu-2、MS、LD、MD的DAT阳性表达物在伤后第2d表达最低;CPu-1、VDB、HDB、CPL、CA1、CA2、CA3、VMH和杏仁核的DAT阳性表达物在伤后第4d表达最低;Pir的DAT阳性表达物在伤后第8d表达最低;Acbc的DAT阳性表达物在伤后第16d表达最低。各脑区恢复时间各不相同,至24d基本恢复正常。以上结果提示DAT可在大鼠不同脑区广泛表达;脑震荡后其在多个脑区呈下调性改变,提示多巴胺转运体的伤后变化可能参与了脑震荡后的认知障碍。     

多巴胺对大鼠背根神经节神经元GABA-激活电流的影响

用全细胞膜片箝记录技术在大鼠分离背根节神经元上观察预加多巴胺对 GABA-激活电流的调控作用。发现大部分受检细胞 ( 4 0 / 47)对外加 GABA敏感。 10 - 6 ～ 10 - 3mol/ L GABA引起一剂量依赖性、有明显去敏感作用的内向电流。在对 GABA敏感的 40个细胞中 ,多巴胺引起一小的、无去敏感的外向电流 ( 2 6/ 40 ) ,其他无明显反应 ( 14 / 40 )。预加多巴胺 10 - 7～ 10 - 4 mol/ L 3 0 s后再加 GABA有 68% ( 2 7/ 40 )的细胞 GABA-激活电流幅值抑制 ,以多巴胺的浓度 10 - 5 mol/ L的抑制最明显 ( 3 3 .3 % )。     

鱼藤酮对PC12细胞多巴胺相关转运体的影响

探讨鱼藤酮对PC12细胞多巴胺转运体(DAT)和突触囊泡单胺转运体2(VMAT2)的影响。我们将PC12细胞经不同浓度的鱼藤酮处理24h后,利用HE染色观察PC12细胞的形态学变化;利用免疫组织化学法观察不同浓度的鱼藤酮对PC12细胞酪氨酸羟化酶(TH),DAT和VMAT2蛋白表达的影响;利用RT-PCR法检测鱼藤酮对PC12细胞DAT和VMAT2基因表达的影响;通过生化实验检测Na+/K+-ATP酶活性。实验结果显示:鱼藤酮浓度为1.0mmol/L时,PC12细胞出现了明显的形态学改变,胞浆染色不均、胞体呈圆形、细胞核形状不规则、核浆比例增加;TH蛋白阳性细胞数量减少;DAT蛋白和基因表达增加;VMAT2蛋白和基因表达降低,与对照组比较均具有统计学意义(P<0.05),且随着鱼藤酮浓度的增加,变化趋势更为明显。鱼藤酮浓度为1.0mmol/L时,Na+/K+-ATP酶活力与对照组比较显著降低(P<0.05)。因此,DAT和VMAT2的表达异常可能参与了毒性作用。     

人及大鼠基底神经节的三维重构及形态比较

利用计算机三维重构技术建立了人及大鼠基底神经节的三维数字化模型 ,并对两者的形态及核团构成进行了比较解剖学研究。在微型计算机上 ,提取人脑及大鼠脑立体定位图谱中含有基底神经节的连续层面 ,利用通用的三维建模及动画软件 3 DStudio MAX,以三维放样的方法 ,在我国首次建立人及大鼠基底神经节的核团及纤维传导通路的三维数字模型 ,对人及大鼠的基底神经节进行了比较解剖学研究。并利用重构的三维模型建立虚拟现实文件 ,在神经解剖学研究中引入浏览器虚拟现实技术 ,实现了任意拆分、组装及旋转上述核团的功能。本研究显示 ,无论在形态上还是在毗邻关系上 ,人与大鼠的基底神经节的差异不大 ;由于人的直立而导致脑干吻尾轴旋转了一定角度 ,其内部核团的相互毗邻关系也随之发生变化 ,从而异于大鼠基底神经节内部核团的毗邻关系 ;基底神经节各个核团间的投射纤维以尽可能短的路径到达靶核团的相应区域 ,构成了拓扑投射关系的基础。     

偏侧帕金森病大鼠基底神经节亚区GABA能神经元的免疫组织化学研究

为探讨基底神经节各亚区γ-氨基丁酸能神经元在帕金森病时的变化。本研究应用免疫组织化学和图象分析技术对 6-羟基多巴胺脑内注射制备的偏侧帕金森病大鼠模型基底神经节亚区 γ-氨基丁酸能神经元的变化进行了观察和计数。结果发现 ,与假手术对照组比较 ,6-羟基多巴胺损毁侧基底神经节亚区γ-氨基丁酸免疫反应阳性神经元的数量均明显减少 (P<0 .0 5 )。本研究结果提示 ,帕金森病时基底神经节直接回路和间接回路兴奋—抑制失衡 ,可能与 γ-氨基丁酸表达减少有关。     

多巴胺受体两个亚家族对大鼠背根神经节GABAA受体的影响

应用全细菌膜片钳技术在大鼠新鲜分离DRG神经元上观察多巴胺D1、D2受体选择性激动剂SKF38393和R（－）－NPA对GABA受体激活电流的影响。在60个对GABA反应的细胞,预加SKF3839 30s后观察到对GABA－激活电流值的影响：多数为抑制（53／60）,少数为增强（1／60）和无明显作用（6／60）。另外对GABA敏感的50个受检细胞,预加R（－）－NPA 30s后对GABA－激活电流幅值的影响也是多数为抑制（38／50）,少数为增强（1／50）和无明显作用（11／50）。SKF3839及R（－）－NPA对GABA（10^4mol／L)－激活电流的抑制作用为浓度依赖性的,上述抑制可为胞内透析蛋白激酶抑制剂H7所翻转。     

新生期孤养对成年大鼠行为与5-羟色胺转运体表达的影响

目的:探讨新生期孤养(neonatal isolation,NI)对成年大鼠行为与5-羟色胺转运体(5-HTT)表达的影响。方法:新生的SD大鼠随机分为正常对照(non-handled,NH)组与新生期孤养组,新生期孤养组在出生后2-14d每天孤养4h,断乳后常规饲养。12w龄时,进行旷场、高架十字迷宫和强迫游泳试验,酶联免疫法检测血浆促肾上腺皮质激素与皮质酮、海马与下丘脑5-HT的含量,原位杂交法观察中缝核5-HTTmRNA的表达。结果:高架十字迷宫试验中新生期孤养组大鼠出入开放臂的总次数与进入开放臂的比率显著减少;强迫游泳试验中新生期孤养组大鼠悬浮时间显著增多(P0.05)。新生期孤养组大鼠血浆促肾上腺皮质激素与皮质酮显著升高,海马中5-HT含量显著减少,中缝核5-HTTmRNA表达显著增强(P0.05或P0.01)。结论:新生期孤养可导致大鼠在成年时出现抑郁和焦虑样行为,可能与下丘脑-垂体-肾上腺轴功能亢进、中枢5-HT功能低下有关。     

雌激素对大鼠心内神经节神经生长因子表达的影响

目的:观察神经生长因子(NGF)在卵巢切除大鼠心房后壁心内神经节的表达及雌激素的调节作用.方法:实验动物分正常组、卵巢切除组、卵巢切除后雌二醇替代组,各组动物审温下饲养3周后,进行 NGF 免疫组织化学显色.结果:各组大鼠心内神经节均存在 NGF 阳性神经元,该类神经细胞胞体多数呈圆形或椭圆形,大、中、小型细胞均有发现.正常组和卵巢切除后雌二醇替代组 NGF 免疫阳性神经无约占全部神经节细胞总数的比例分别为 61%和64%,但卵巢切除组大鼠心内神经节 NGF 阳性神经元的数量明显减少,表达明显降低,NGF 免疫阳性细胞约占全部神经节细胞总数的比例为28%.结论:心内神经节细胞内含 NGF;雌激素可能影响心内神经节细胞的 NGF 表达.     

神经内镜额部锁孔手术与显微手术治疗基底节区出血的临床疗效

目的探讨神经内镜额部锁孔手术与显微手术治疗基底节区出血的治疗效果。方法回顾性分析徐州医科大学附属医院神经外科2017年8月至2019年6月收治的34例经手术治疗基底节区出血患者的临床资料。根据手术方式分为内镜额部锁孔手术组(观察组)14例和显微手术组(对照组)20例,比较2组患者术前、术中和术后的病情变化及手术并发症发生情况,总结两种手术方式的临床应用效果。结果观察组平均血肿清除率、手术时间均少于对照组,差异均有统计学意义(P<0.05);2组患者术后并发症发生率比较,差异无统计学意义(P>0.05);观察组术后不同时间GCS评分及术后3个月ADL评分均优于对照组,差异有统计学意义(P<0.05)。结论与显微手术相比,神经内镜额部锁孔手术具有手术时间短、血肿清除率高、术后并发症少、患者预后较好等优点。     

基底节梗塞与出血对记忆影响的研究

采用《临床记忆量表》研究了９１例基底节梗塞与出血患者的记忆损害状况。研究表明：（１）基底节梗塞与出血病人的记忆商数（ＭＱ）约６４％在中等水平以下，与对照组比较差异显著；（２）基底节出血组的记忆障碍比基底节梗塞组更为严重；（３）四项与词有关的分测验及ＭＱ均以基底节左侧损害组下降明显，联想学习尤为明显，呈现大脑功能一侧化现象；（４）基底节梗塞或出血其损害体积的大小与记忆障碍的程度无明显正相关。     

Effect of neuroleptics on acetylcholine metabolism in the basal ganglia

Neuroleptics (haloperidol, clozapine, pimozide, chlorpromazine) caused a decrease in the level of the free (functionally active) form of acetylcholine (ACh) and also, to some extent, of the bound form of ACh, inconsistent changes in the content of loosely bound (vesicular) form of ACh, and a weak effect on choline acetyltransferase activity in the basal ganglia of rats 5–30 min after injection. By contrast with their inhibitory action on acetylcholinesterase (AChE) activity, in experiments in vitro most neuroleptics (except clozapine) increased ACh activity in vivo. These results show that neuroleptics activate ACh metabolism and evidently stimulate cholinergic structures in the basal ganglia; AChE activity can be used as a criterion of this stimulating action of the neuroleptics.Problem Laboratory in Neurohormone Biochemistry, Minsk Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR V. V. Zakusov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 4, pp. 429–431, April, 1978.     

The contribution of synaptic plasticity in the basal ganglia to the processing of visual information

A mechanism for the involvement of the basal ganglia in the processing of visual information, based on dopamine-dependent modulation of the efficiency of synaptic transmission in interconnected parallel associative and limbic cortex-basal ganglia-thalamus-cortex circuits, is proposed. Each circuit consists of a visual or prefrontal area of the cortex connected with the thalamic nucleus and the corresponding areas in different nuclei of the basal ganglia. The circulation of activity in these circuits is supported by the recurrent arrival of information in the thalamus and cortex. Dopamine released in response to a visual stimulus modulates the efficiencies of “strong” and “weak” corticostriatal inputs in different directions, and the subsequent reorganization of activity in the circuit leads to disinhibition (inhibition) of the activity of those cortical neurons which are “strongly” (“weakly”) excited by the visual stimulus simultaneously with dopaminergic cells. The pattern in each cortical area is the neuronal reflection of the properties of the visual stimulus processed by this area. Excitation of dopaminergic cells by the visual stimulus via the superior colliculi requires parallel activation of the disinhibitory input to the superior colliculi via the thalamus and the “direct” pathway” in the basal ganglia. The prefrontal cortex, excited by the visual stimulus via the mediodorsal nucleus of the thalamus, mediates the descending influence on the activity of dopaminergic cells, simultaneously controlling dopamine release in different areas of the striatum and thus facilitating the mutual selection of neural reflections of the individual properties of the visual stimulus and their binding into an integral image. __________ Translated from Zhurnal Vysshei Nervnoi Deyatel’nosti imeni I. P. Pavlova, Vol. 56, No. 6, pp. 742–756, November–December, 2006.     

神经生长因子对体外培养大脑基底神经节神经元的保护作用

目的:研究神经生长因子(NGF)对谷氨酸造成的体外培养大脑基底神经节神经元损伤后的再生及防止神经元坏死的保护作用。方法:原代培养的胎鼠前脑基底神经节神经元,分为对照组、谷氨酸损伤组和谷氨酸损伤后NGF保护组。用倒置相差显微镜进行活细胞观察、采用RT-PCR技术检测前脑基底神经节神经元GAP-43和NF-L基因表达。结果:谷氨酸损伤神经元胞体回缩,突起消失或断裂。加入NGF后神经元绝大多数细胞胞体饱满,突起明显,细胞问的网络联系清晰可见,接近于对照组;NGF保护组大脑基底神经节GAP-43 mRNA和NF-L mRNA表达与损伤组比较具有显著性差异。结论:NGF可保护谷氨酸造成体外培养的大脑基底神经节损伤后的再生,并防止神经元坏死。     

Movement disorders and lesions of pigeon brain stem analogues of basal ganglia

The nucleus spiriformis lateralis (SpL) and the nucleus tegmenti pedunculo-pontinus, pars compacta (TPc) of the pigeon are thought to be analogous to the mammalian substantia nigra. The analogy is based upon comparative neuroanatomical and histochemical similarities. To test the analogy on a behavioral basis, unilateral electrolytic or kainate lesions were produced in selected nuclei of the diencephalon and mesencephalon. Birds demonstrated behaviioral disturbances similar to those induced in mammals after lesions to the substantia nigra. The most obvious disturbance was a persistent turning in a direction opposite to the side of the lesion. The birds also demonstrated postural problems and arhythmic movements of the head or whole body tremor. The similarities in movement disorders strengthen on a behavioral basis, the suggested analogies between the mammalian substantia nigra and the SpL and TPc. The pigeon may be another useful experimental model to study movement disorders associated with damage to the basal ganglia.     

神经生长因子对谷氨酸介导的基底核神经元损伤的保护作用

本研究的目的是观察神经生长因子(nerve growth factor,NGF)对谷氨酸诱导的基底核神经元损伤的保护作用。取出生后1d乳鼠前脑基底核神经元培养,随机分为正常对照组、模型组(谷氨酸损伤组)和NGF保护组。用倒置相差显微镜进行活细胞观察,采用RT-PCR技术检测前脑基底核神经元的神经生长相关蛋白-43(growth associated protein-43,GAP-43)的表达。结果显示谷氨酸损伤组神经元胞体回缩,突起消失或断裂。NGF保护组的神经元绝大多数胞体饱满,突起明显,细胞间的网络联系仍清晰可见,接近于正常对照组;NGF保护组神经元内GAP-43mRNA表达比谷氨酸损伤组高,两者比较有统计学意义(P<0.05)。结果提示,NGF能保护基底核神经元免受谷氨酸兴奋性毒性的损伤。     

Unilateral 6-hydroxydopamine lesion of dopamine neurons and subchronic L-DOPA administration in the adult rat alters the expression of the vesicular GABA transporter in different subsets of striatal neurons and in the substantia nigra, pars reticulata

The loss of dopamine neurons combined or not with the subsequent administration of L-DOPA in patients with Parkinson's disease or in experimental models of the disease results in altered GABAergic signaling throughout the basal ganglia, including the striatum and the substantia nigra, pars reticulata. However, the molecular mechanisms involved in altered GABA neurotransmission remain poorly understood. In order to be released from synaptic vesicles, newly synthesized GABA is transported from the cytosol into synaptic vesicles by a vesicular GABA transporter. The objective of this study was to examine the hypothesis that expression of the vesicular GABA transporter (vGAT) is altered in the unilateral 6-hydroxydopamine model of Parkinson's disease. Our results provide evidence that a unilateral 6-hydroxydopamine lesion results in increased and decreased vGAT mRNA levels in striatopallidal and striatonigral neurons, respectively. These two subsets of neurons were identified by the co-expression or lack of co-expression of preproenkephalin, a marker of striatopallidal neurons, using double-labeling in situ hybridization histochemistry. Such changes occurred in the striatum ipsilateral to the 6-hydroxydopamine lesion and were paralleled by decreased vGAT protein levels in the substantia nigra, pars reticulate (SNr). On the other hand, the subchronic systemic administration of L-DOPA increased vGAT mRNA levels in preproenkephalin-negative neurons on the side ipsilateral and, to a lesser extent, the side contralateral to the 6-hydroxydopamine lesion. Systemic L-DOPA also increased vGAT protein levels in the ipsi- and contralateral SNr. As a whole, the results provide original evidence that vGAT expression is altered in the 6-hydroxydopamine model of Parkinson's disease. They also suggest that the behavioral effects induced by a subchronic administration of L-DOPA to 6-hydroxydopamine-lesioned rats involve an increase in the vesicular release of GABA by striatonigral neurons.     

Projections of the basal ganglia to the zona incerta of the dog diencephalon

Retrograde axonal transport of horseradish peroxidase was used to show that the projections of the globus pallidus, entopeduncular nucleus, substantia nigra, and pedunculopontine tegmental nucleus in dogs are directed to all segments of the zone incerta. The experiments reported here identified no topical features in the organization of these projections in dogs, as application of marker to different areas of the zona incerta yielded similar distributions of labeled neurons in the basal ganglia. No striatal projections to the zone incerta were found.