抑制一氧化氮合成酶对大鼠睡眠及5—羟色胺神经元免疫反应的影响

采用多导描记及免疫组化方法，观察了侧脑室和腹腔给予硝基左旋精氨酸对慢性植入电采大鼠睡眠量及中缝背核５－羟色胺神经元免疫阳性反应的影响，结果表明：Ｌ－ＮＮＡ显著抑制慢波睡眠的快眼动睡眠，与对照相比较，觉醒时间延长。     

脑缺血时中缝背核5-HT变化的免疫组化观察

目的：探讨沙土鼠有离缺血时中缝背核神经元５－ＨＴ的变化。方法：用免疫组织化学ＡＢＣ法及图象分析对沙土鼠缺血不同时间以及缺血１０分钟后再灌不同时间其中缝背核神经元６－ＨＴ的变化进行研究。结果：脑缺血１０钟、３０分钟时,沙土鼠脑中缝背核５－ＨＴ阳性神经元的平均光密度值（ＯＤ）值与对照组比较无显著性差异;而缺血４小时时则明显降低,差异有显著性（Ｐ〈０．０５）,脑缺血１０分钟后,再灌流２０分钟,５－ＨＴ下     

大鼠中缝背核向缰核投射的5—羟色胺能神经元对躯体伤害性刺激的fo …

用ＷＧＡ－ＨＲＰ逆行追踪与抗ＦＯＳ和抗５－羟色胺免疫组化的三重标记方法，观察向大鼠一侧有前跖部皮下注射８％福尔马林５０μｌ后，中缝背核与缰核５－羟色胺神经元的ＦＯＳ表达，光镜下发现中缝背核内有七种阳性神经元，即ＨＲＰ，ＰＯＳ，５－ＨＴ单标细胞，ＦＯＳ／ＨＲＰ，ＦＯＳ／５－ＨＴ，ＨＲＰ／５－ＨＴ，双标细胞ＦＯＳ／ＨＲＰ／５－ＨＴ三标细胞，结果表明中缝背核至缰核的５－ＨＴ能投射神经元对躯体的伤害性刺激     

大鼠孤束核内5－羟色胺能纤维的来源──HRP逆行追踪与5－羟色胺免疫组织化学双标记洁研究

用ＨＲＰ注入孤束核逆行追踪和５－羟色胺（５－ＨＴ）免疫组织化学相结合的双标记法，观察大鼠孤束核内的５－ＨＴ能纤维和终末的来源。结果在脑干内见到许多ＨＲＰ单标、５－ＨＴ单标和ＨＲＰ／５－ＨＴ双标细胞，其中ＨＲＰ／５－ＨＴ双标细胞主要分布于脑桥被盖核（２６．５７％）、脑桥网状核（２４．１９％）、中缝大核（２１．１７％），其次为中缝隐核（１２．７４％）、中缝苍白核（７．７８％），少数见于中缝背核、中脑导水管周围灰质和中缝脑桥核等处。而在结状神经节内仅见到ＨＲＰ单标细胞，未见到５－ＨＴ单标和ＨＲＰ／５－ＨＴ双标细胞。以上结果表明，大鼠孤束核的５－ＨＴ能纤维和终末主要来源于脑桥与延髓的５－ＨＴ神经元的投射。本研究提示脑桥与延髓内上述核团的５－ＨＴ神经元通过向孤束核的直接投射参与孤束核内脏功能的调控。     

大鼠脑干5-羟色胺能和儿茶酚胺能神经元向孤束核的投射——荧光金标记结合免疫荧光技术研究

应用荧光金逆行追踪和免疫荧光技术相结合的方法,对投射至大鼠孤束核的５－羟色胺能和儿茶酚胺能神经纤维的脑干来源进行了研究。结果证明,将荧光金注入一侧孤束核的中尾段,逆标神经元分布于脑干下行抑制系统的大部分核团。结合５－羟色胺和酪氨酸羟化酶免疫荧光组织化学技术研究发现：荧光金／５－ＨＴ 双重反应阳性神经元分布于中脑导水管周围灰质的腹外侧区、中缝背核、脑桥被盖网状核、脑桥尾侧网状核、中缝桥核、中缝正中核、中缝大核、巨细胞网状核α部、中缝隐核和中缝苍白核等核团,其中中央灰质腹外侧区、中缝大核、巨细胞网状核α部、中缝隐核、中缝苍白核等处的荧光金／５－ＨＴ 双重反应阳性神经元数量占脑干向孤束核投射的５－ＨＴ 阳性神经元总数的近７０％ （６９．９１％ ）;荧光金／酪氨酸羟化酶双重反应阳性神经元主要分布于脑干中脑中央灰质腹外侧区、Ａ７、蓝斑及蓝斑下核（Ａ６）、Ａ５、巨细胞网状核α部和Ａ１ 等核团,其中中央灰质腹外侧区、Ａ５、巨细胞网状核α部和Ａ１ 内的荧光金／酪氨酸羟化酶神经元数量占脑干向孤束核投射的酪氨酸羟化酶阳性神经元总数的８８．１７％ 。本研究提示,大鼠脑干下行抑制系统形成了至ＮＴＳ中尾段的以中脑中央灰质腹外侧区及中缝大核、?     

“更年健”对老年雌性大鼠中缝背核5—羟色胺的影响

应用免疫组织化学和计算机图像分析技术,观察中药“更年健”Ｃ地老龄雌性大鼠中缝背核５－ＨＴ的影响。结果表明：“更年健”和苯甲酸雌二醇用药后,中缝背核５－羟色胺含量明显增高,与用药相比,相关显著。提示“更年健”能增强老年雌性大鼠中缝背核５－ＨＴ神经元活性。     

大鼠孤束核内5—羟色胺能纤维的来源—HRP逆行追踪与5—羟色…

用ＨＲＰ注入孤束核逆行追踪和５－羟色胺免疫组织化学结合的双标记法，观察大鼠孤束核内的５－ＨＴ能纤维和终末的来源，结果在脑干内见到许多ＨＲＰ单标，５－ＨＴ单标和ＨＲＰ／５－ＨＴ双标细胞，其中ＨＲＰ／５－ＨＴ双标细胞主要分布于脑桥被盖核（２６．５７％），脑桥网状核（２４．１９％）、中缝大核（２１．１７％），其次为中缝隐核（１２．７４）、中缝苍白核（７．７８％），少数见于中缝背核、中脑导水管周围灰质和中     

豚鼠中脑中缝背核和中缝正中核5—羟色胺能神经元向视交叉上核的直…

实验借助ＨＲＰ逆行定位和免疫细胞化学定性相结合的双合的双重标记技术，探讨豚鼠中脑中缝背核和中缝正中核向正丘脑视交叉上核的直接投射。结果首次确定了（１）中缝背核／中缝正中核内５－羟色胺和非５－羟色胺能神经元对式叉上核的直接投射和分布特点；（２）投射神经元主要集中集中背核；（３）中缝背核对视交叉上核的投射神经元几乎均为５－羟色胺能，而缝中中５－色胺和非５－羟色胺能投射神经元的各占半数。     

大鼠脑干面口部运动神经元与5-HT样、SP样、L-ENK样、GABA样和Gly样阳性终末间的突触联系

为了观察５－羟色胺（５－ＨＴ）样、Ｐ物质（ＳＰ）样、亮氨酸－脑啡肽（Ｌ－ＥＮＫ）样、γ－氨基丁酸（ＧＡＢＡ）样和甘氨酸（Ｇｌｙ）样阳性终末与脑干面口部运动核（三叉神经运动核、面神经核和舌下神经核）间的突触联系，用ＨＲＰ逆行追踪与免疫组织化学染色相结合的双标技术，在电镜下对５－ＨＴ样、ＳＰ样、Ｌ－ＥＮＫ样、ＧＡＢＡ样和Ｇｌｙ样阳性终末与ＨＲＰ逆标面口部运动神经元间的突触联系进行了观察。５－ＨＴ样、ＳＰ样、Ｌ－ＥＮＫ样、ＧＡＢＡ样和Ｇｌｙ样阳性终末在脑干面口部运动核内呈均匀分布，没有区域特异性。将ＨＲＰ注入面口部肌肉或相应的周围神经后，ＨＲＰ逆标神经元见于脑干运动核不同部位内支配相应肌肉的运动神经元。５－ＨＴ样、ＳＰ样和Ｌ－ＥＮＫ样阳性终末主要与ＨＲＰ逆标运动神经元的树突形成非对称性的轴－树突触；ＧＡＢＡ样和Ｇｌｙ样阳性终末则主要与ＨＲＰ逆标运动神经元的树突形成对称性的轴－树突触；偶见上述阳性轴突终末与ＨＲＰ逆标运动神经元的胞体形成轴－体突触。结果提示：主要来自脑干运动前神经元的５－ＨＴ样、ＳＰ样和Ｌ－ＥＮＫ样阳性终末及ＧＡＢＡ样和Ｇｌｙ样阳性终末可能分别对脑干运动神经元发挥兴奋性或抑制性调控作用     

大鼠脑于5—羟色胺能和儿茶酚胺能神经元向孤束核的投射——荧光 …

应用荧光金逆行追踪和免疫荥光技术相结合的方法,对投射至大反束核的５－羟色胺能和儿茶酚受能神经纤维的脑睐源进行了研究。结果证明,将荧光金注入一侧孤束核的中尾段,逆标神经元分布于脑干下行抑制系统的大部分核团。结合５－羟色胺和酷按酸羟化酶免疫荧光组织化学技术研究发现：荧光金／５－ＨＴ双重反应阳性神经元分布于中脑导水管周围灰质的腹外侧区、中缝背核、脑桥补盖网状核、脑桥尾侧网状核、中缝桥核、中缝正中核、巨细     

多重脑震荡大鼠脑干中缝核团内5-羟色胺免疫阳性表达变化的研究

为了探讨多重脑震荡(multiple cerebral concussion,MCC)后大鼠中缝核团内5-羟色胺(5-HT)能神经元的变化规律,本实验采用自制单摆式机械打击装置复制MCC大鼠模型,研究伤后大鼠脑干中缝核团内5-HT及5-HT合成过程中的限速酶-色胺酸羟化酶(TPH)的表达。将56只大鼠随机分为7组:对照组、伤后1、2、4、8、16和24d组(n=8)。用免疫组织化学染色技术及图像分析法定量分析伤后大鼠脑干中缝核团内5-HT和TPH的表达变化。结果显示:(1)TPH免疫反应阳性产物在中缝背核、正中中缝核的表达在伤后2d时达到高峰,与正常对照组相比有显著性差异(P<0.05);中缝大核和中缝苍白核分别以伤后1d组和4d组阳性反应最强;(2)5-HT免疫反应阳性产物在中缝背核、正中中缝核的表达也在伤后2d时达到高峰,16、24d组基本恢复至正常水平;而中缝大核和中缝苍白核内5-HT的免疫反应性在各损伤组与正常对照组之间均无显著性差异(P>0.05)。以上结果表明,多重脑震荡后中缝核团内TPH和5-HT的表达增高,这为研究5-HT对MCC后认知障碍的影响提供了形态学依据。     

Distinguishing characteristics of serotonin and non-serotonin-containing cells in the dorsal raphe nucleus: electrophysiological and immunohistochemical studies

The membrane properties and receptor-mediated responses of rat dorsal raphe nucleus neurons were measured using intracellular recording techniques in a slice preparation. After each experiment, the recorded neuron was filled with neurobiotin and immunohistochemically identified as 5-hydroxytryptamine (5-HT)-immunopositive or 5-HT-immunonegative. The cellular characteristics of all recorded neurons conformed to previously determined classic properties of serotonergic dorsal raphe nucleus neurons: slow, rhythmic activity in spontaneously active cells, broad action potential and large afterhyperpolarization potential. Two electrophysiological characteristics were identified that distinguished 5-HT from non-5-HT-containing cells in this study. In 5-HT-immunopositive cells, the initial phase of the afterhyperpolarization potential was gradual (tau=7.3+/-1.9) and in 5-HT-immunonegative cells it was abrupt (tau=1.8+/-0.6). In addition, 5-HT-immunopositive cells had a shorter membrane time constant (tau=21.4+/-4.4) than 5-HT-immunonegative cells (tau=33.5+/-4.2). Interestingly, almost all recorded neurons were hyperpolarized in response to stimulation of the inhibitory 5-HT(1A) receptor. These results suggested that 5-HT(1A) receptors are present on non-5-HT as well as 5-HT neurons. This was confirmed by immunohistochemistry showing that although the majority of 5-HT-immunopositive cells in the dorsal raphe nucleus were double-labeled for 5-HT(1A) receptor-IR, a small but significant population of 5-HT-immunonegative cells expressed the 5-HT(1A) receptor. These results underscore the heterogeneous nature of the dorsal raphe nucleus and highlight two membrane properties that may better distinguish 5-HT from non-5-HT cells than those typically reported in the literature. In addition, these results present electrophysiological and anatomical evidence for the presence of 5-HT(1A) receptors on non-5-HT neurons in the dorsal raphe nucleus.     

Evidence for a role of GABA interneurones in the cortical modulation of midbrain 5-hydroxytryptamine neurones.

Recent electrophysiological studies demonstrate that the ventral medial prefrontal cortex has a powerful inhibitory influence on 5-hydroxytryptamine (5-HT) neurones in the dorsal raphe nucleus. Here we utilised a combination of anatomical and electrophysiological methods to characterise the cellular substrate underlying this effect.Anterograde tracing (Phaseolus vulgaris leucoagglutinin) using electron microscopy demonstrated a pathway from the ventral medial prefrontal cortex that makes neuronal contacts throughout the dorsal raphe nucleus. These contacts were predominantly asymmetrical synapses adjoining GABA immunoreactive dendrites and spines.In vivo extracellular recordings were made in the dorsal raphe nucleus of the anaesthetised rat from a subpopulation of non-5-HT neurones. These neurones were fast-firing, irregular and with short spike width, properties strongly reminiscent of immunochemically identified GABA interneurones in other brain regions. Recordings of classical 5-HT neurones were also included. Electrical stimulation of the ventral medial prefrontal cortex elicited a rapid onset (16 ms latency), orthodromic excitation of the non-5-HT neurones (13/25 neurones). This stimulation also caused a pronounced inhibition of most 5-HT neurones tested, with a longer latency (30 ms), and this was partially blocked by locally applied bicuculline.These data provide the first evidence that the ventral medial prefrontal cortex influences the activity of large numbers of raphe 5-HT neurones by targeting a local network of GABA neurones. This circuitry predicts that physiological and pathological changes in the ventral medial prefrontal cortex will impact on significant parts of the forebrain 5-HT system.     

PTSD样行为异常大鼠中缝背核5-HT1A受体的改变

目的观察创伤后应激障碍(PTSD)样行为异常大鼠中缝背核(DRN)神经元5-HT1A受体表达变化,为探讨PTSD的发病机制提供资料。方法成年健康雄性Wistar大鼠60只,随机分为连续单一刺激(SPS)模型1d、4d、7d组及正常对照组,采用免疫组化、Western blot方法检测PTSD样行为异常大鼠中缝背核神经元5-HT1A受体表达变化。结果 SPS刺激后1d大鼠中缝背核神经元5-HT1A受体表达水平开始逐渐升高,第4天高于第1天,第7天高于第4天。结论 PTSD样大鼠中缝背核神经元5-HT1A受体呈规律性过表达。     

大鼠中缝背核接触脑脊液神经元化学性质的研究

用CB-HRP追踪与免疫细胞化学结合的方法,对大鼠中缝背核接触脑脊液神经元(简称触液神经元)的化学性质进行了研究.将CB-HRP注入第三脑室后,中缝背核内观察到CB-HRP标记细胞,标记细胞分布于中缝背核的背侧部和腹侧部.在CB-HRP与P物质或5-羟色胺免疫细胞化学结合的切片上,中缝背核内出现三种标记细胞:HRP单标细胞,P物质或5-羟色胺免疫反应阳性单标细胞,HRP/SP或HRP/5-HT双标细胞,双标细胞为中、小型梭形或圆形细胞.上述结果提示:中缝背核存在SP能和5-HT能触液神经元.     

Origin,distribution, and morphology of serotonergic afferents to the cat superior colliculus: a light and electron microscope immunocytochemistry study

Summary We have studied the serotonergic (5-HT) projection to the cat superior colliculus (SC) using serotonin antibody immunocytochemistry and retrograde transport of peroxidase-conjugated wheatgerm agglutinin (WGA-HRP). In 3 experiments, the two labels were combined in order to double label cells with both anti-5-HT and WGA-HRP. In the remaining experiments, the two labels were examined separately. Serotonin-like immunoreactive fibers were found throughout all layers of SC, but were most densely distributed within the zonal and upper superficial gray layers. Most 5-HT fibers were thin and had characteristic varicosities and terminal swellings. At the EM level, immunoreactive terminals and varicosities were found to contain small agranular vesicles and occasionally large granular vesicles (LGVs). Conventional synaptic densities were only rarely observed. Injections of WGA-HRP into SC resulted in labeling of neurons throughout the dorsal raphe nucleus and surrounding ventrolateral periaqueductal gray. Only a few cells were found in the raphe medianus and raphe pontis and none within the raphe magnus or other medullary raphe nuclei. Cells in the dorsal raphe giving rise to the SC projection varied in shape, size, and morphology and must represent more than one cell type. The morphology of these cells was indistinguishable from that of cells in the dorsal raphe which were double labeled by anti-5-HT and WGA-HRP. We conclude that the 5-HT innervation of the superior colliculus varies in density in different laminae, arises from several different cell types, and originates primarily from the dorsal raphe nucleus with minor projections from raphe medianus and raphe pontis.     

双轴旋转运动刺激后大鼠中缝大核神经元活动分析

目的:分析大鼠经引发晕动病的双轴旋转运动刺激后,中缝背核(DR)内5-HT能神经元激活状况,以探讨5-HT水平与前庭刺激的关系.方法:将雄性SD大鼠随机分成两组:对照组和双轴旋转运动刺激组.所有动物装入定做的有机玻璃圆筒内,并于黑暗中进行如下处理2h:运动刺激组动物以电动马达驱动的双轴旋转运动刺激;对照组动物放置在距刺激仪器20 cm的位置.应用包含DR的冠状脑切片,以双重免疫荧光标记技术标记5-HT和Fos蛋白,并对标记神经元进行计数和统计分析.结果:对照组和旋转运动刺激组的Fos阳性神经元、5-HT阳性神经元以及Fos/5-HT双标记神经元的数目分别是:93.4 ±12.0 vs 153.1 ±21.1、528.5±36.0vs 531.1±23.4和15.1 ±11.3 vs 30.8 ±11.3.t检验分析显示两组Fos阳性神经元数目有显著性差异(P＜0.05);Fos/5-HT双标神经元数目在旋转运动刺激后有增加趋势.结论:引发晕动病的双轴旋转运动能激活中缝背核内的神经元,其中的5-HT能神经元也对刺激有反应.     

Individual cells in the raphe nuclei of the medulla oblongata in rat that contain immunoreactivities for both serotonin and enkephalin project to the spinal cord

Summary The ventral medulla oblongata of rats was analyzed with a double-labelling immunofiuorescence technique using guinea pig antibodies directed against serotonin (5-HT) and rabbit antisera directed against enkephalin (ENK). Numerous cells in the region of nucleus raphe obscurus, nucleus raphe pallidus and nucleus raphe magnus showed immuno-staining for either 5-HT or ENK. A substantial number of cells showed positive immunostaining for both 5-HT and ENK. 5-HT/ENK double-labelled cells were most frequently encountered in an area that extended from the rostral aspect of the inferior olivary nucleus to the pontomedullary border. This region corresponds anatomically to nucleus raphe magnus/nucleus paragigantocellularis. In addition, a number of the 5-HT/ENK-containing cells were retrogradely labelled with Fluoro-Gold dye that had been injected into the thoracic spinal cord several days prior to perfusion. Schematic drawings showing the anatomical distribution of 5-HT/ENK colocalization are provided.     

Ipsilateral and contralateral serotonergic projections from dorsal and median raphe nuclei to the forebrain in rats: immunofluorescence quantitative analysis

The objective of this study was to clarify the 5-HT projections from the right and left sides of the dorsal (DRD), ventral (DRV) and lateral (DRL) subdivisions of the middle level of the dorsal (DR) raphe nucleus and median (MR) raphe nucleus to the lateral septum (LS), preoptic area (POA) or ventromedial hypothalamus (VMH), which are important neural substrates for neuroendocrine regulation of reproduction. A retrograde neural tracer, Fluoro-Gold (FG), was infused into the right side of these regions in ovariectomized rats and the numbers of FG and/or 5-HT immunopositive cells in the right and left sides of the raphe nuclei were counted. It was found that the POA and VMH received more 5-HT projections than the LS from the DR and MR. In the subdivisions of the DR, 70% of all 5-HT projections from the DR to these 3 areas originated from the DRL. Furthermore, ipsilateral projections from the DR to the POA and VMH but not to the LS were dominant, compared to the contralateral projections. A right–left difference was not seen among the MR 5-HT projections. Thus, laterality of the projections is thought to be strong in the 5-HT clusters located far from the midline of the midbrain raphe nuclei.