5-HT_1受体亚型mRNAs在坐骨神经分支选择损伤模型大鼠脊髓背角的表达变化

为探讨脊髓内5-HT1受体亚型在神经病理性痛信息传递和调控中的作用,本研究利用反转录聚合酶链式反应(RT-PCR)技术观察了坐骨神经分支选择性损伤(SNI)模型大鼠脊髓背角内5-HT1A、5-HT1B、5-HT1D、5-HT1E和5-HT1F受体亚型mR-NAs表达变化。结果表明:损伤侧脊髓背角内5-HT1A受体亚型mRNA的表达水平在术后3d明显增高,7d达到高峰,随后开始下降,术后28d与正常对照相比仍有显著变化;5-HT1B受体亚型mRNA的表达水平也于术后3d开始显著增高,此后持续升高,至21d达到高峰,28d时有轻微下调,但高于对照水平;未观察到损伤侧脊髓背角内5-HT1D受体亚型mRNA表达水平的显著变化。5-HT1F受体亚型mRNA的表达水平于术后4d显著增高,此后一直维持该表达水平至28d。上述受体亚型在非损伤侧脊髓背角内的表达水平均无变化。在脊髓背角内未检测到5-HT1E受体亚型mRNA的表达。上述5-HT1受体亚型mRNA在SNI模型脊髓背角内具有不同的表达变化特点,提示不同的5-HT1各受体亚型在神经病理性痛信息的传递和调节中可能发挥着不同的作用。     

5-HT_(2~7)受体亚型mRNAs在坐骨神经分支选择性损伤模型大鼠背根神经节的表达变化

本研究利用反转录 聚合酶链式反应(RT- PCR)方法,观察了坐骨神经分支选择性损伤(SNI)所致神经病理性痛条件下,大鼠背根神经节(DRG)中 5- HT2~7受体亚型mRNAs的时程表达变化。用RT- PCR方法在正常大鼠DRG中检测到 5-HT2A、5- HT3、5- HT4、5 -HT5A和 5 HT7受体亚型mRNAs的表达,但未检测到 5 -HT2B、5- HT2C、5- HT5B和 5 -HT6受体亚型mRNAs。SNI能诱导 5 -HT2A、5- HT3、5 -HT4 和 5- HT7受体亚型mRNAs在损伤侧DRG的表达上调。其中, 5- HT2A受体亚型mRNA的表达在术后 3d时开始升高,持续增加至 28d; 5- HT3 受体亚型mRNA的表达在术后 4d时明显增加, 14d时达到高峰; 5- HT4受体亚型mRNA于术后 3d的表达明显增加, 21d时达到高峰; 5-HT7 受体亚型mRNA的表达在术后 1d时即显著升高,一直维持高水平的表达至28d。未检测到 5- HT5A受体亚型mRNA的表达变化。在SNI对侧的DRG,各受体亚型mRNAs的表达未出现明显变化。部分 5- HT受体亚型在SNI模型DRG的表达具有不同的时程变化特点,提示它们在SNI所致的神经病理性痛中发挥着不同的作用。     

补肾中药对老年大鼠脑组织中胆碱能M受体分布的影响

采用受体放射配基结合分析、原位杂交和放射自显影术等方法,研究胆碱能M受体、M1受体mRNA在脑组织中的分布,观察补肾中药对胆碱能M受体的含量、功能及分布的影响,探讨补肾中药延缓脑组织衰老的机理.结果显示:老年大鼠大脑皮层、海马、纹状体中M受体明显减少,放射自显影的灰度值降低,胆碱能M1受体mRNA在老年大鼠纹状体部位的表达减少,在其他部位的不明显.补肾中药明显增加老年大鼠胆碱能M受体的数量,对脑内胆碱能M1受体mRNA转录水平降低的部位有改善作用.补肾中药可通过改善胆碱能M1受体的mRNA表达、增加脑内胆碱能M1受体数量来延缓脑组织衰老.     

雌二醇对去卵巢大鼠下丘脑5-HT_(1A)和5-HT_(2A)受体亚型mRNA表达的影响

观察17β-雌二醇(17β-estradioI,E2)对去卵巢大鼠下丘脑5-HT1A和5-HT2A受体mRNA表达的影响的时间效应。实验动物分为假去卵巢对照组(OVX+S组)和去卵巢组(OVX)。去卵巢组大鼠又分为雌激素处理组(OVX+E+组)和无雌激素处理组(OVX+E-组),分别在处理后3、6、9和12 d时,采用反转录-聚合酶链式反应(RT-PCR)方法观察5-HT1A和5-HT2A受体mR-NA在大鼠下丘脑的表达。结果显示:5-HT1A和5-HT2A受体mRNA在所有大鼠下丘脑均有表达;5-HT1A受体mRNA在3、6和9 d的OVX+E-组大鼠下丘脑的表达水平明显高于同时间点的OVX-E+组,而5-HT2A受体mRNA在6、9和12 d的OVX+E-组下丘脑的表达量也显著高于同时间点的OVX+E+组。本文结果表明去卵巢后大鼠下丘脑5-HT1A和5-HT2A受体亚型mRNA表达水平升高,17β-雌二醇可下调两种受体的表达。     

雌激素对大鼠心脏雌激素受体α和β表达的影响

目的研究雌激素受体（ER）α和β在新生、成年和卵巢切除大鼠心脏的表达。方法采用半定量逆转录聚合酶链反应（RT-PCR）和Western blot方法,分析新生和成年大鼠心脏各部的ERα和βmRNA及ER蛋白质的表达,探讨不同剂量的17β雌二醇对卵巢切除大鼠心脏ERα和βmRNA、ER蛋白质的表达调控及对心房、心室重量和它们与体重比变化的影响。结果ERα mRNA在新生大鼠心脏各部表达较低,而成年大鼠心脏各部ERα mRNA的转录水平提高3～20倍,这种差别体现在ERα蛋白质的表达水平不同。反之新生大鼠心脏ERβ mRNA表达水平较高,而成年大鼠心脏ERβ mRNA的表达降低2～7倍。卵巢切除减少成年大鼠心房ERαmRNA和蛋白质的表达,减轻心房重量及心房／体重比;17肛雌二醇替代却能反转这些改变,同时随着血浆17β-雌二醇水平的增高,这种影响在一定程度上呈剂量依赖关系。卵巢切除和178一雌二醇替代并不改变心脏和心室／体重比,也不影响ERβ mRNA的表达。结论雌激素受体α和β在新生和成年大鼠心脏的表达存在差别,ERβ可能在大鼠心脏的发育过程中起重要作用,ERα足雌激素对成年大鼠心脏调控的优势受体;雌激素的主要靶器官是心房,且通过其受体的介导促进心房肌细胞的增生。     

5-HT_1受体各亚型mRNAs在大鼠脊髓不同节段背角和腹角的表达(英文)

为进一步了解5-HT受体在中枢感觉和运动机能中的作用,本研究利用反转录PCR方法比较了5-HT1A,5-HT1B,5-HT1D,5-HT1E和5-HT1F受体亚型mRNAs在脊髓不同节段的背角、腹角的表达。结果显示:5-HT1A,5-HT1B,5-HT1D受体亚型mRNA的表达水平在胸、腰、骶段脊髓的背角明显高于腹角;而颈段背角内的表达仅略高于腹角。脊髓内未检测到5-HT1E受体亚型;5-HT1F受体亚型在脊髓各节段的背角和腹角都有较高水平的表达。5-HT1受体家族各亚型mRNA在脊髓背角和腹角表达水平的差异可能提示它们在中枢感觉和运动功能中发挥不同的作用。     

5-HT受体亚型mRNAs在大鼠背根神经节和三叉神经节的表达—原位杂交组织化学法研究

应用原位杂交组织化学技术观察了大鼠背根节和三叉神经节内 5 -HT2 、5 -HT3 、5 -HT4、5 -HT5 、5 -HT6 和 5 -HT7受体亚型 m RNAs的表达。结果显示 :在背根节和三叉神经节中均观察到了 5 -HT2 A、5 -HT3 、5 -HT4和 5 -HT7受体亚型 m RNAs阳性细胞 ;两种神经节内上述受体亚型 m RNAs阳性细胞的百分比不同。 5 -HT3 受体亚型 m RNA阳性细胞的百分比最高 ,分别为 3 0 .4% (背根节 )和 3 2 .8% (三叉神经节 )。约 17.2 %和 14 .6 %的背根节和三叉神经节细胞分别呈 5 -HT2 A受体 m RNA阳性 ;而 5 -HT4和 5 -HT7受体亚型 m RNAs阳性细胞在此二节中的百分比分别为 14 .9%、2 4.7%和 11.2 %、9.8%。各受体亚型 m RNA阳性细胞均以中、小型节细胞为主。在各神经节内均未观察到 5 -HT2 B、5 -HT2 C、5 -HT5 和 5 -HT6 受体 m RNA的表达。上述结果提示 5 -HT2 A、5 -HT3 、5 -HT4和 5 -HT7受体亚型在外周伤害性感受中可能发挥着重要的作用     

大鼠纹状体边缘区内5-HT_(2A)受体的分布──原位杂交和免疫组织化学研究

已知纹状体边缘区有密集的 5 -HT纤维及终末分布。为了观察 5 -HT2 A受体 m RNA是否在大鼠纹状体边缘区表达 ,从基因分子水平和细胞水平进一步证明大鼠纹状体边缘区能否合成 5 -HT2 A受体 ,用地高辛标记的寡核苷酸探针进行原位杂交 ,研究了大鼠纹状体边缘区的 5 -HT2 A受体 m RNA的表达及分布。原位杂交结果发现 5 -HT2 A受体 m RNA阳性杂交信号在纹状体的分布不均匀 ,尾壳核只有少量中等大小的阳性胞体 ,苍白球也只有少量较大的阳性胞体 ,而在尾壳核和苍白球之间的边缘区部位则可见到许多中等大小的梭形阳性神经元胞体 ,并呈现密集的带状分布。免疫组织化学结果观察到 5 -HT受体阳性神经元胞体在纹状体的分布与原位杂交结果一致。本实验结果推测 ,大鼠纹状体边缘区神经元可以合成 5 -HT2 A受体 ,具有接受和整合 5 -HT神经递质的功能     

携带大鼠酪氨酸羟化酶基因慢病毒载体的构建及转染

目的:构建携带增强绿色荧光蛋白(EGFP)和大鼠酪氨酸羟化酶(TH)基因的慢病毒载体,转染大鼠骨髓间充质干细胞(rMSC),观察TH基因的表达.方法:RT-PCR方法获得大鼠TH基因,将其克隆至慢病毒载体.通过瞬时转染法包装出病毒上清,鉴定滴度.感染rMSCs,荧光显微镜下观察EGFP的表达、转染效率,RT-PCR、免疫印迹法分别检测TH mRNA和蛋白的表达情况.结果:重组慢病毒载体质粒pNL-TH-IRES2-EGFP经双酶切鉴定正确,所获TH基因经测序后与GenBank报道序列完全一致;生产的病毒浓缩后滴度为4.1×10~7TU/ml;感染rMSCs结果显示荧光激发可见绿色荧光,TH-rMSCs、空载体-rMSCs组5d转染效率差异无统计学意义.RT-PCR、免疫印迹法显示TH基因成功在rMSCs中表达.结论:成功构建带有EGFP和大鼠TH基因的慢病毒载体,并获得TH-rMSCs基因工程细胞.     

栀子苷抑制甲型流感病毒感染细胞中Toll样受体7/核因子-κB信号通路

目的 研究栀子苷对甲型流感病毒H3N2感染所致的Toll样受体7(Toll-like receptors7,TLR7)及其介导的细胞内转录因子核因子-κB(nuclear factor-kappa B,NF-κB)活性及前炎症细胞因子TNF-α和IL-6释放的影响,以探讨栀子苷干预甲型流感病毒作用的分子生物学机制.方法 甲型流感病毒H3N2作为刺激因素,利用双荧光素酶顺式报告系统和免疫荧光实验检测栀子苷对NF-κB转录活性及NF-κB核转位的影响.RT-PCR法进一步验证栀子苷对NF-κB上下游靶基因TLR7、TNF-α和IL-6 mRNA表达水平的影响.结果 通过NF-κB双萤光素酶报告系统检测发现,与正常对照组比较,病毒感染的细胞中NF-κB荧光素酶报告活性明显升高;与病毒损伤组比较,栀子苷治疗组NF-κB荧光素酶报告活性明显受到抑制.荧光倒置显微镜下观察NF-κB p65磷酸化水平及核转位变化结果显示,病毒感染的细胞中NF-κB磷酸化水平及人核率明显升高;与病毒损伤组比较,栀子苷治疗组NF-κB磷酸化水平及入核率明显受到抑制.RT-PCR结果显示栀子苷能明显抑制病毒诱导的TLR7、TNF-α和IL-6 mRNA的高表达.结论 栀子苷可拮抗甲型流感病毒H3N2感染细胞中TLR7介导的细胞内转录因子NF-κB信号转导通路的活化,并可抑制其下游靶基因TNF-α和IL-6 mRNA的高表达水平来发挥抗病毒感染的作用.     

Asymmetry of the internal connections of the striate cortex of the cat in the projection zone of the center of the field of vision

Studies were carried out on the organization of the internal connections of the striate cortex in cats in the projection zone of the center (0–5°) of the field of vision by microintophoretic application of horseradish peroxidase to electrophysiologically identified orientational columns. The area containing neurons showing retrograde labeling in most cases extended in the mediolateral direction. Labeled cells were located in the upper (II, III) and lower (V, VI) layers of the cortex, and the shapes and orientations of the areas containing labeled neurons in these layers coincided. Spatial asymmetry was detected in the distribution of labeled neurons relative to the orientational column studied. Labeled cells were located predominantly medial to the columns, regardless of the distance from the projection of the area centralis. Considering the visuotopical map of field 17, the asymmetry detected here provides evidence that neurons in orientational columns have more extensive connections with neurons of the peripheral part of the cortex. An asymmetrical distribution of “silent” zones around the receptive fields of neurons in orientational columns is suggested, and that these appear to receive influences from the periphery of the visual field. Laboratory of Visual Physiology and Laboratory of Central Nervous System Morphology, I. P. Pavlov Institute of Physiology, Russian Academy of Sciences, 6 Makarov Bank, 199034 St. Petersburg, Russia. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 82, No. 12, pp. 23–29, December, 1996.     

Effects of the lesion of the postcommissural part of the septum on the behavior of the rat.

The effects of the lesion of the postcommissural part of the septum on behavior of the rat has been studied. Results may be summarized as follows. An increase in the exploratory behavior in the open field which decreases rapidly; a decrease in the number of defecations in this test and a decrease in time leaving a dark environment for exploration. In the shuttle box test, no facilitation of the acquisition, but a permanent and quite significant increase in the intertrial activity has been found. We conclude that the lesions tend to decrease the emotivity of the subjects. An interpretation on the basis of the species -- specific defensive reactions explains the transitory and permanent effects of the lesions on the spontaneous activity.     

The development of the ligament of the head of the femur

The hip joints of 30 human male and female fetuses and stillborns between 20 mm and 350 mm crown-rump length were studied by light microscopy. The ligament of the head of the femur developed in situ as a condensation of mesenchyme at the end of the second month of intra-uterine life (IUL), and was vascularized by branches of acetabular vessels early in the fourth month. In the majority of fetuses older than 5.5 months IUL, vessels in the ligament passed a short way into the femoral head within cartilage canals, to supply a small region around the fovea capitis. The remainder of the head was supplied by vessels in canals from around the upper part of the neck. The ligament changed from predominantly cellular to fibrous during the last 4 months of IUL. This increase in strength suggested significant mechanical functions in utero: limitation of adduction-flexion and opposition to postero-superior dislocation were the most likely.     

Counterpointing the functional role of the forebrain and of the brainstem in the control of the sleep-waking system

This paper reviews the lifetime contributions of the author to the field of sleep-wakefulness (S-W), reinterprets results of the early studies, and suggests new conclusions and perspectives. Long-term cats with mesencephalic transection show behavioral/polygraphic rapid eye movement sleep (REMS), including the typical oculo-pupillary behavior, even when the section is performed in kittens prior to S-W maturation. REMS can be induced as a reflex. Typical non-rapid eye movement S (NREMS) is absent and full W/arousal is present only after a precollicular section. The isolated forebrain (IF) rostral to the transection exhibits all features of W/arousal and NREMS [with electroencephalographic (EEG) spindles and delta waves], arousal to olfactory stimuli, and including the appropriate oculo-pupillary behaviors. These features also mature normally after neonatal transection. REMS is absent from the IF. After deprivation there is NREMS pressure and rebound in the IF, but the decerebrate cat only shows pressure for REMS. Most IF reactions to pharmacologic agents are within expectations, except for the tolerance/withdrawal effects of long-term morphine use which are absent. In contrast, these effects are supported by the brainstem (i.e. seen in the decerebrate cat). In cats with ablation of the telencephalon, or diencephalic cats, delta waves are absent in the thalamus. EEG thalamic spindle waves are seen triggering S for only 4-5 days after ablation. Therefore, true NREMS is absent in chronic diencephalic cats although pre- and postsomniac behaviors persist. These animals are hyperactive and show a pronounced, permanent insomnia; however, a low dose of barbiturate triggers a dramatic REMS/atypical NREMS rebound. Cats without the thalamus (athalamic cats), initially show a dissociation between behavioral hyperactivity/insomnia and the neocortical EEG, which for 15-20 days exhibits only delta and slower oscillations. Fast, low-voltage W rhythms appear later on, first during REMS, but spindle waves and S postures are absent from the start, such that these cats also display only atypical NREMS. Athalamic cats also show barbiturate-sensitive insomnia. Cats with ablation of the frontal cortices or the caudate nuclei remain permanently hyperactive. They also show a mild, but significant hyposomnia, which is permanent in afrontal cats, but lasts for about a month in acaudates. The polygraphic/behavioral features of their S-W states remain normal. We conclude and propose that: (a) the control of the S-W system is highly complex and distributed, but is organized hierarchically in a well-defined rostro-caudal manner; the rostral-most or highest level (telencephalon), is the most functionally complex/adaptative and regulates the lower levels; the diencephalic/basal forebrain, or middle level, has a pivotal role in inducing switching between S and W and in coordinating the lowest (brainstem) and highest levels; (b) W can occur independently in both the forebrain and brainstem, but true NREMS- and REMS-generating mechanisms exist exclusively in the forebrain and brainstem, respectively; (c) forebrain and brainstem S-W processes can operate independently from each other and are preprogrammed at birth; this helps understanding normal and abnormal polygraphic/behavioral dissociations in humans and normal dissociations/splitting in aquatic mammals; (d) NREMS homeostasis is present in the IF, but only REMS pressure after deprivation persists in the decerebrate cat; (e) the thalamus engages in both NREMS and W; (f) insomnia in diencephalic cats is the result of an imbalance between antagonistic W- and S-promoting cellular groups in the ventral brain (normally modulated by the telencephalon); (g) the EEG waves, which are signature for each S-W state, appear to truly drive the concomitant behaviors, e.g. a hypothetical human IF could alternate between behavioral NREMS and W/arousal/awareness; (h) a role for REMS is to keep the individual sleeping at the end of the self-limiting NREMS periods. The need for accelerating research on telencephaling NREMS periods. The need for accelerating research on telencephalic S-W processes and downstream control of the lower S-W system levels is emphasized.     

Topical organization of the projections of neurons of the substantia nigra and the ventral field of the tegmentum of the midbrain to the head of the caudate nucleus of the cat

Neuroscience and Behavioral Physiology -     

Involvement of the striatum in the central regulation of the hormonal functions of the gonads

Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms. Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 85, No. 4, pp. 594–597, April, 1999.     

Ultrastructure of the endothelium of the drainage system of the eye

The endothelium of the ocular drainage system (Schlemm’s canal, collector tubules, and aqueous veins) in primary juvenile glaucoma undergoes degenerative dystrophic changes with compensatory hypertrophy and proliferation at the initial stages of the glaucomatous process and atrophy and desquamation at advanced and terminal stages. Progressive decrease in the pinocytous function of endotheliocytes, reduction of the protein-synthesizing and mitochondrial compartments of the cytoplasm, and formation of autophagosomes reflect the process of endotheliocyte degeneration in general. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 145, No. 5, pp. 574–577, May, 2008