妊娠中期奶山羊下丘脑中催产素的免疫组化定位

采用免疫组织化学技术中的链霉素抗生物素-过氧化酶(streptarid in-perox idase,SP)法,对妊娠中期奶山羊下丘脑中催产素(O xytoc in,OT)神经元的定位及分布进行了观察研究。结果表明,下丘脑中分泌OT的神经元主要分布在视上核和室旁核,在环核、视上弥散核、弓状核、室周核、穹窿周核、下丘脑前区、下丘脑外侧区、下丘脑后核和乳头体各核团等也有一定数量的阳性神经元;另外,在室旁核、视上弥散核、视上核、正中隆起和第三脑室附近有较多数量的强阳性神经纤维,在交叉上核有少量阳性神经纤维。提示OT在妊娠中期奶山羊下丘脑中分布广泛,且下丘脑OT经神经垂体,第三脑室,血管三条途径释放,经血液循环最终作用于靶细胞而发挥生理作用。     

催产素样免疫反应神经元在鸡下丘脑的分布──ABC法研究

ＡＢＣ法研究结果表明，催产素（ＯＴ）样神经元在鸡下丘脑中分布于一条前端尖细、中央宽厚、后端钝圆的长梭形细胞带内，带的前端达前联合核、皮质联合床核，后端达乳头体各核团，中央部分包括视上核、室旁核、视交叉上核、腹内侧核、腹外侧核、背内侧核、背外侧核、弓状核、室周核等，ＯＴ样神经纤维主要分布在第三脑室室周和正中隆起的中间带和外侧带。通过ＯＴ释放途径的观察，对ＯＴ在体内的含量变化进行了讨论     

乏情期奶山羊下丘脑催产素样神经元的分布

为了揭示不同生理期催产素(Oxytocin,OT)在下丘脑-垂体-性腺轴中的分布特点,采用链霉素抗生物素蛋白-过氧化物酶(Streptaridin-peroxidase,SP)法研究了OT免疫反应产物在乏情期奶山羊下丘脑中的分布。结果显示:OT样神经元存在于视上核、室旁核弓状核、室周核、穹隆周核、下丘脑前核、下丘脑后核、腹外侧核、背内侧核、下丘脑外侧区以及乳头体各核团等27个核区,分布区域呈前端尖、中部宽、后钝圆的梭形带。此外,在室旁核观察到大量的强阳性神经纤维,第三脑室附近的纤维伸向脑室内。说明OT在下丘脑广泛存在,且下丘脑OT经神经垂体、第三脑室、血管3条途径释放,经血液循环最终作用于靶细胞而发挥生理作用。     

兔脑和脊髓内开胃素A免疫阳性神经元和神经纤维的分布

采用免疫组织化学法研究了10只青紫蓝兔脑内开胃素（Orexin） A免疫阳性神经元和神经纤维的分布。结果显示,Orexin A免疫阳性神经元分布于下丘脑的视上核、室旁核、背内侧核、穹隆周核、外侧区、前区和后区以及底丘脑的未定带。Orexin A免疫阳性神经纤维广泛分布于中枢神经系统内,在端脑分布于大脑皮质、尾状核、隔核和杏仁核;在间脑分布于丘脑、下丘脑、上丘脑和垂体;在中脑分布于中央灰质、前丘、后丘、黑质、网状结构和中缝核;在脑桥分布于蓝斑、网状结构和中缝核;在延髓分布于极后区、孤束核和迷走神经背侧运动核;在小脑和脊髓也有分布。     

GnRH神经元在发情期奶山羊下丘脑中的免疫组化定位

为了研究促性腺激素释放激素(GnRH)在发情期奶山羊下丘脑中的表达特点,采用超敏的免疫组化链霉素抗生物素蛋白-过氧化物酶法研究GnRH免疫反应神经元在发情期山羊下丘脑中的分布。结果显示,GnRH免疫反应阳性细胞主要分布在视上核、视前内侧核、视前外侧核、交叉上核、丘脑外侧区、室旁核后部、弓状核、腹外侧核、结节乳头体核、乳头体内侧核、乳头体外侧核等核团。阳性细胞的形态有圆形、三角形、卵圆形、梭形、多角形、不规则形,有些阳性细胞还具有明显的突起。圆形和三角形细胞直径的大小为5~35μm,卵圆形和梭形细胞短轴和长轴大小分别为5~27μm、15~58μm。免疫反应阳性产物还分布于部分纤维束中,如视束、正中中隆起等。根据细胞中阳性反应的强弱可分为强阳性,如交叉上核;中等阳性,如乳头体内侧核;弱阳性,如前穹窿周核。上述研究结果表明,GnRH在成年奶山羊下丘脑广泛存在。     

Immunoreactive dynorphin-(1-8) and corticotropin- releasing factor in subpopulation of hypothalamic neurons

Immunoreactive corticotropin-releasing factor (CRF) and dynorphin-(I-8) were visualized in rat hypothalamus by immunohistofluorescence with specific antibodies. In brains from colchicine-treated, adrenalectomized rats, neuronal perikarya with immunoreactive CRF were observed in the paraventricular nucleus of the hypothalamus. The CRF occurred together with the dynorphin-(1-8). However, the CRF immunoreactivity occurred only in a subpopulation of the dynorphin-(1-8) immunoreactive cells. These findings suggest that there may be a functional interrelationship of CRF with dynorphin-related opioid peptides and provide further evidence that neurons may contain more than one bioactive substance.     

Orexin A在鹌鹑间脑内定位分布的SABC法研究

采用免疫组织化学SABC法,研究orexin A在鹌鹑间脑内的分布。结果显示,orexinA阳性神经元胞体主要分布在间脑中下丘脑区的室旁核、室周区、外侧核和后内侧核,其阳性纤维则投射到丘脑和下丘脑的广大区域,其中在圆核、三角核、前脑外侧束、第五额束、视顶盖、中央白质层和外侧核等处较密集。结果表明orexin A存在于鹌鹑的间脑内,但其分布与大鼠之间有明显的差异。     

雌激素对老年大鼠小脑ER、ChAT、NGF表达的影响

 【目的】 探讨雌激素对大鼠小脑内雌激素受体（ER）、神经生长因子（NGF）和胆碱乙酰转移酶（ChAT）表达的影响。【方法】运用超敏感的免疫组织化学SP法，以老年SD大鼠小脑为研究对象，通过补充17β－雌二醇对ER、NGF和ChAT在小脑中的表达和分布变化进行研究。【结果】ER、NGF和ChAT免疫阳性反应物分布于小脑的蒲肯野氏细胞层、小脑齿状核、小脑间位核和小脑室顶核，ER阳性产物主要定位于细胞胞浆和突起中，也存在于胞膜和胞核中。老年大鼠小脑皮质及小脑核中ER、NGF和ChAT的表达强度及阳性细胞数量总趋势是显著降低，而补充17β－雌二醇后三种阳性产物的强度和阳性细胞数目显著回升，蒲肯野氏细胞的阳性突起长度和数量也呈此变化趋势。【结论】雌激素可促进NGF和ChAT的表达，在维持和保护小脑神经元的结构和功能中发挥了重要作用；另外ER、NGF和ChAT表达变化的相似性提示三者在雌激素对小脑的作用中是相互调节和影响的，同时表明雌激素在小脑发挥作用可能既通过基因组机制，也通过非基因组机制途径。     

神经元在动物下丘脑中分布的研究进展

对近年来研究较多的神经元在动物下丘脑各个神经核中的分布情况作了介绍,指出神经元在下丘脑各个神经核均有分布,甚至有几种神经元同时分布在某些神经核中,视上核(SON)、室旁核(PVN)及弓状核(ARC)是多种神经元集中分布的主要神经核,为进一步研究下丘脑的生理功能提供了形态学资料。     

Effects of Estrogen on ER, NGF, and ChAT Expression in Cerebellum of Aging Female Sprague-Dawley Rat

This article discusses the effects of estrogen on the expression of estrogen receptor （ER）, nerve growth factor （NGF）, and choline acetyltransferase （CHAT） in the cerebellum of rats. The model of aging female rat was established to study the expression and distribution of ER, NGF, and ChAT in the cerebellum following 17β-estradiol treatment using the technique of immunohistochemical ultrasensitive SP in sprague-dawley rat. The immunoreactive productions were distributed in stratum Purkinje cell, nucleus dentatus, nucleus interpositus, and nucleus fastigii of cerebellum, and the ER positive production was mainly located in the plasma, cytoplasmic membrane, and neurite, and also existed in nucleus. The general tendency of the expression of ER, NGF, and ChAT positive production in the cerebellum cortex and nuclei of aging rat significantly decreases, while the intensity and quantity of the immunoreactive production ascends predominantly after 17β-estradiol treatment. Simultaneously, the positive neurite of Purkinje cell shows a similar tendency. The above- mentioned results suggest that the estrogen upregulates the expression of NGF and CHAT, and plays a vital role in sustaining and protecting the structure and function of cerebellum neurons. Furthermore, the similarity of their changing tendency implies that they were correlated and cooperated during the course in effect of estrogen on cerebellum. It also showed that the action of estrogen in cerebellum could be via genomic and nongenomic mechanism.     

柄海鞘神经复合体的显微观察

采用甲苯胺蓝染色和Golgi镀银法对柄海鞘神经复合体的结构进行显微观察,并通过免疫组织化学方法对甲硫氨酸脑啡肽在神经复合体中的分布进行了研究。①甲苯胺蓝染色法显示,神经复合体由神经节、神经腺和疏松结缔组织组成;神经节分为皮质部、髓质部和过渡带;神经腺位于神经节背侧面,由腺小叶组成;神经细胞呈深蓝色,神经纤维呈浅蓝色,与背景反差较大。Golgi镀银法显示的结果不理想,神经细胞和神经纤维着色不明显。②免疫组化结果显示,M-ENK在神经复合体中分布广泛,神经细胞、神经纤维及上皮细胞均有Met-ENK阳性反应。     

猪下丘脑开胃素 A的分布定位

用免疫组织化学方法研究了5头苏钟猪下丘脑内开胃素(orexin)A的分布。结果表明,在猪下丘脑内,开胃素A免疫阳性神经元分布于下丘脑的视前内侧区、室周核、室旁核、视上核、背内侧核、穹隆周核、乳头体核、前区、外侧区和后区等部位,以下丘脑外侧区、乳头体核和视上核出现的免疫阳性神经元最多,以室周核的最少。这一结果与在其他动物上获得的基本相似。     

A major direct GABAergic pathway from zona incerta to neocortex

Retrograde fluorescent tracers were used to demonstrate a previously unknown but sizable direct gamma-aminobutyric acid (GABA)-containing neuronal pathway from the zona incerta to the neocortex in rats. This incertocortical pathway was found to project bilaterally to the entire neocortex and exhibited a rough corticotopic organization. Many of the zona incerta neurons projecting to the parietal and occipital cortices could also be immunohistochemically stained with antibodies to glutamic acid decarboxylase and GABA. Few of these neurons were immunoreactive to tyrosine hydroxylase antibodies, which identify dopamine-containing neurons. Injections in the frontal and entorhinal cortices labeled many neurons near or within the dopaminergic A13 subdivision of the zona incerta. In addition, the incertocortical system was found to be significantly larger during early postnatal (2 to 3 weeks) development. The projection pattern of this newly discovered pathway resembles that of the monoaminergic and cholinergic systems, arising from the brainstem and forebrain, suggesting possible similarities of function.     

Immunocytochemical detection of acetylcholine in the rat central nervous system

A specific antibody to acetylcholine was raised and used as a marker for cholinergic neurons in the rat central nervous system. The acetylcholine conjugate was obtained by a two-step immunogen synthesis procedure. An enzyme-linked immunosorbent assay was used to test the specificity and affinity of the antibody in vitro; the results indicated high affinity. A chemical perfusion mixture of allyl alcohol and glutaraldehyde was used to fix the acetylcholine in the nervous tissue. Peroxidase-antiperoxidase immunocytochemistry showed many acetylcholine-immunoreactive cells and fibers in sections from the medial septum region.     

神经干细胞定向诱导分化条件研究

通过模拟体内分化诱导条件,在体外对神经干细胞(NSCs)进行定向诱导.结果表明,由添加2% B27 100 pg·mL-1 IL-1 0.05 g·L-1 Vc 5 u·mL-1 EPO的DMEM/F12的诱导液A和1份含2% B27的DMEM/F12 1份纹状体提取液混合组成的诱导液B均可以将中脑和大脑皮质的NSCs诱导分化为多巴胺能神经元,TH阳性细胞表达率随诱导时间的增加而提高,中脑来源的NSCs的TH阳性细胞诱导率极显著地高于大脑皮质来源的NSCs,诱导液A的诱导效果优于诱导液B.     

Adrenergic innervation of the parasympathetic ciliary ganglion in the chick

The chick ciliary ganglion receives a nonvascular symathetic innervation in addition to the well-known cholinergic one; fluorescent, varicose adrenergic fibers form pericelluar baskets. Adrenergic fibes were identified electron microscopically in ganglia fixed with potassium permanganate. The fibers degenerate after injection of 6-hydroxydopamine. No true synaptic relationships involving adrenergic varicosities and ganglion cells or cholinergic terminals were demonstrable. The distribution of the adrenergic fibers suggests a kind of "distance à synapse" with the choroidal cells or with the preganglionic fibers (or both). The adrenergic innervation might provide a modulation of the cholinergic transmission.     

人参三醇型皂苷黑根霉发酵产物与人参皂苷的增强记忆活性对比

通过建立3种损伤性拟痴呆模型,即东莨菪碱致胆碱能损伤拟痴呆小鼠模型、40%乙醇损伤拟痴呆小鼠模型、氯霉素注射致痴呆小鼠模型,每日给予小鼠受试药物,通过跳台试验和被动回避试验对小鼠进行行为学检测,并对小鼠脑内胆碱能含量进行检测,对比人参三醇型皂苷的黑根霉发酵产物和部分人参皂苷的增强记忆活性。结果表明:人参三醇型皂苷黑根霉发酵产物3种损伤模型中均显示出增强记忆活性,且与对照组相比差异显著。人参三醇型皂苷黑根霉发酵产物组和人参皂苷Rg1组小鼠脑内乙酰胆碱转移酶(ChAT)活性明显高于空白对照组(P0.05),乙酰胆碱酯酶(AchE)活性低于空白对照组(P0.05),说明人参三醇型皂苷黑根霉发酵产物不仅能降低AchE活性,而且能增强ChAT活性,并且此活性与小鼠脑内的胆碱能系统具有一定关系。     

鸡INR的神经束与血－神经屏障的超微结构

 【目的】研究鸡肠Remak神经（INR）神经束的超微构造与血－神经屏障的微细形态,为进一步阐明INR的生理功能提供理论基础。【方法】应用透射电镜技术研究鸡肠Remak神经（INR）神经束的超微构造与血－神经屏障的微细形态。【结果】鸡INR被神经束膜分隔成许多神经束,束内既有无髓纤维又有有髓纤维,但以无髓神经纤维的数量占绝对优势。雪旺氏细胞与两种神经纤维的形成与结构密切相关。神经束膜细胞突起长而薄,包绕神经束,其丰富的粗面内质网、线粒体和发达高尔基复合体,以及以常染色质为主的细胞核和明显的核仁结构,提示束膜细胞具有活跃的蛋白合成和分泌功能。不同于肠壁内ENS缺乏神经屏障的现象,鸡INR的神经束膜中分布有血－神经屏障,是由结构紧密的毛细血管及其周围多层排列的束膜细胞形成的,且束膜细胞表面可见吞饮泡的形成,但未见不同束膜细胞之间有特殊的细胞连接。这种组成特点表明,鸡INR的血－神经屏障没有哺乳动物外周神经的严密。【结论】上述试验结果显示,鸡INR的细胞学结构既不同于肠壁内ENS,也与外周神经有差异,具有一定的特殊性。     

Atriopeptin-immunoreactive neurons in the brain: presence in cardiovascular regulatory areas

Antisera to atriopeptin III and to a cyanogen bromide fragment of the precursor molecule atriopeptigen were prepared and used to examine the distribution of atriopeptin-like immunoreactive material in the heart and brain of the rat. Granules of this material were seen in myocytes throughout the right and left atria and were densest in the perinuclear region. The distribution of atriopeptin-like immunoreactive material in the heart is consistent with previous reports of atrial secretory granules. In the brain neurons containing the material were observed in the hypothalamus and the pontine tegmentum. Atriopeptin in the brain may serve as a neurotransmitter in neural systems controlling blood volume and composition, the same physiological functions regulated by blood-borne atriopeptin.     

广西鸡源H9N2亚型禽流感病毒分离鉴定及遗传进化分析

[目的]明确广西鸡源H9N2亚型禽流感病毒(AIV)的分子遗传变异规律及其对公共卫生安全的影响,为H9亚型禽流感(AI)的防控提供参考依据.[方法]从广西某肉鸡养殖场采集病料,经SPF鸡胚接种进行病毒分离,对初步鉴定为H9亚型AIV分离株的HA、NA、PB2、PB1、PA、NP、M和NS等8个基因片段进行克隆测定及遗传进化分析,同时对分离毒株的生物学特性进行测定.[结果]从广西发病鸡群中分离获得一株H9N2亚型AIV,命名为A/chicken/Guangxi/BT1/2016(H9N2).分离毒株A/chicken/Guangxi/BT1/2016(H9N2)HA基因的裂解位点为PSRSSR↓GLF,具有典型的低致病性AIV(LPAIV)分子特征,在遗传进化关系上属于4.2.5分支,与国内常用H9N2亚型AIV疫苗株所属的4.2.3分支存在一定差异;其他7个基因也分别来源于不同的AIV亚型.分离毒株A/chicken/Guangxi/BT1/2016 (H9N2)同时具备与α-2,3唾液酸和α-2,6唾液酸受体结合的特性,且具有感染人类的潜在风险;其感染鸡群后病毒分布主要集中在气管、肺脏和脾脏,可通过呼吸道和消化道同时排毒,且在攻毒后第1d即开始排毒,第5d为排毒高峰.[结论]从广西发病鸡群中分离获得的A/chicken/Guangxi/BT1/2016(H9N2)属于LPAIV,是由不同亚型AIV重组产生的H9N2亚型新毒株,可导致鸡群疫苗免疫失败,且具有感染人类的潜在风险.