电脉冲刺激肾旁穴对兔丘脑雌激素受体蛋白表达的影响

采用电脉冲刺激穴位法及免疫组织化学SP法，研究了电脉冲刺激肾旁穴对丘脑内雌激素受体免疫反应阳性产物的影响。结果发现雌激素受体免疫反应阳性产物在丘脑内广泛分布，可见电针组丘脑室旁核、丘脑腹外侧核、丘脑腹内侧核、丘脑腹主核、丘脑中央中核、丘脑网状核、丘脑室周核等核区有大量阳性产物，且多分布于神经元的胞浆、胞核和突起中、有少部分分布于胞膜上，染色较深，突起明显，细胞形态多样，轮廓清晰；而对照组相应神经核团中阳性细胞数量较少且淡染，突起也不明显，形态单一或轮廓不清。两组阳性产物的染色强度和细胞数目均差异显著。以上结果表明电脉冲刺激肾旁穴可使家兔上述核团内雌激素受体的表达增强。     

ER免疫反应产物在幼龄公山羊下丘脑-垂体-性腺轴的表达

应用免疫组化SP法时雌激素受体(ER)免疫反应产物在幼龄公山羊下丘脑、垂体、性腺中的分布特点进行了研究。结果显示，下丘脑中ER免疫反应阳性神经元主要分布在视上核、室旁核、室周核等9个核团，在视前区、下丘脑外侧区等核团也有一定数量的阳性神经元；阳性细胞呈圆形、卵圆形、三角形不等，阳性物质大多位于细胞质和胞核，阳性纤维散布于各阳性核团中；在正中隆起和第三脑室室管膜可见大量小而密集排列的深染的阳性神经元。神经垂体中可见大小不等、排列较均匀且染色较深的纤维，腺垂体中腺细胞呈强阳性着色。睾丸曲细精管中的初级精母细胞和支持细胞以及睾丸间质细胞中的ER阳性产物均为微弱表达。由此表明，幼龄公山羊雌激素除作用于性腺外，还主要作用于中枢神经系统的广泛区域，推测其参与了脑中生殖、内分泌、认知等多种功能的调控。     

Leptin长形受体mRNA与LHRH在猪前脑内共存

为了寻找Leptin在下丘脑水平调控生殖内分泌的形态学证据，用原位杂交和免疫组织化学相结合的方法，研究了5头三元杂交仔猪前脑内Leptin长形受体（OB-Rb）mRNA和黄体生成素释放激素（LHRH）免疫反应阳性物质的共存关系。结果表明，OB-Rb mRNA和LHRH免疫反应阳性神经元在下丘脑、海马结构、大脑皮层和杏仁核内共存，即上述结构内一些表达Leptin长形受体mRNA的神经元也含有LHRH免疫反应阳性物质。在下丘脑，双阳性神经元主要位于室旁核、室周核、腹内侧核、外侧区、弓状核；在大脑皮层（额叶和顶叶）内，双阳性神经元主要位于Ⅲ～Ⅴ层；在海马结构内，双阳性神经元主要位于齿状回的多形层和颗粒层以及海马的锥体细胞层。结果提示，Leptin可能通过其受体直接作用于下丘脑的促性腺激素释放激素（GnRH）神经元调节动物的生殖和内分泌活动。     

突触素在鸡产蛋前后下丘脑内生殖核团的表达和发育

应用免疫组织化学方法研究了突触素 (SVP- 38)免疫反应产物在鸡产蛋前后下丘脑中生殖内分泌核团内的表达和发育情况。结果显示 ,前联合区、视前室旁核、室周弓状核、下丘脑室旁核、乳头体内侧核、乳头体外侧核等生殖核团内突触素免疫反应强度 ,在 3个年龄组间差异显著。 2 40 d组反应强度最高 ,70 d组次之 ,5 0 0 d组最低。结果表明 ,产蛋高峰期下丘脑内的突触可塑性增强 ,突触素产蛋高峰期表达强度最高 ,后随日龄增加而强度下降。说明突触素的表达与生殖内分泌激素的分泌有依赖性。     

雌激素受体α在雌性大鼠脑中的表达

本试验利用免疫组织化学SP法,研究了雌激素受体α在大鼠脑中的表达.结果表明,ER_α免疫反应产物主要定位于神经元细胞核,部分区域出现阳性神经元和阳性突起,在海马CA1、CA2辐射层、齿状回、下托的分子层等出现阳性星形胶质细胞;ER_α在脑中表达广泛,免疫反应产物在端脑的隔外侧核、海马锥体细胞层、大脑皮质等,间脑的缰内侧核、下丘脑室旁核、弓状核、视上核等,中脑的动眼神经核,脑桥的脑桥核外侧部等为高密度;在端脑的尾壳核、杏仁中央核、海马CA4区,间脑的下丘脑室周核、缰外侧核等,中脑的黑质致密带,脑桥的脑桥被盖核等为较高密度;在问脑的视前室周核、丘脑后核、中脑的红核、脑桥的脑桥被盖网状核等为中等密度;在端脑的苍白球,间脑的无名质,无免疫反应产物出现.结果显示,雌激素在调节脑的认知和生殖内分泌过程中,ER_α可能起主要作用;雌激素可通过ER_α调节星形胶质细胞功能.     

鸡脑催乳素免疫反应神经元的分布

运用ＡＢＣ法观察了催乳素（ＰＲＬ）免疫反应神经元在鸡脑的分布。结果,ＰＲＬ阳性胞体主要分布在视交叉上核、视前室旁核、视上核、下丘脑室旁核、弓状核和伏隔核,旧纹状体、正中隆起存在大量阳性纤维末梢,在侧脑室腹侧的室管膜和脑基底神经胶质板上也存在ＰＲＬ阳性神经元。视前室旁核、视交叉上核、下丘脑室旁核、弓状核等核团内的ＰＲＬ阳性神经元有突起向第三脑室投射,伏隔核内及侧脑室室管膜上的ＰＲＬ阳性神经元有突起伸至侧脑室,视上核的ＰＲＬ阳性神经元也有突起投射至脑基底神经胶质板,表明鸡脑内的ＰＲＬ可以释放入脑室系统,参与调节脑－脑脊液神经体液回路。     

IFN-γR和ER在大鼠下丘脑中的共存

为了探讨神经-免疫-内分泌网络中生物活性物质间相互调节和平衡的关系,本试验应用免疫组化双标记法对成年SD雌性大鼠下丘脑中IFN-γ受体（IFN-γR）和雌激素受体（ER）的共表达进行了研究。结果发现,IFN-γR和ER广泛存在于大鼠下丘脑中,其中在视前交叉上核、视前内侧核、室周核、交叉上核、下丘脑前核、下丘脑内侧核、下丘脑外侧核、乳头体前背侧核、乳头体前腹侧核等13个核团中存在大量的IFN-γR和ER双标记细胞,双标记细胞约占全部阳性标记细胞的60．9％;双标记细胞胞质呈黑褐色、胞核呈棕黄色;ER单标着色细胞以神经胶质细胞居多。研究结果表明,IFN-γ和雌激素可以分别以其各自受体为介质进行信号传递和信息整合,同时也通过在同一细胞中的相互作用而参与机体的神经-免疫-内分泌调节。     

鸡端脑和下丘脑向面神经腹侧核的传出性纤维联系—HRP逆行追踪法

将２０％ＨＲＰ水溶液注射于鸡的面神经腹侧核，逆行追踪支配该部的有关主同位神经核团。结果于端，下丘脑和脑干内出现标记细胞，端脑内的标记细胞较多地出现于古纹状体，少量出现于旧纹状体腹侧与枕中脑束之间的区域，两群标记细胞形态上明显不同。下丘脑的标记细胞较多地出现于下丘脑后区，室旁核出现少量，个别散在于下丘脑其他区域。     

成年发情期奶山羊下丘脑催产素及其mRNA的表达

应用免疫组织化学SP法和原位杂交法研究了催产素（oxytocin，OT）及OT mRNA在成年发情期奶山羊下丘脑中的分布和表达。结果，OT免疫反应阳性细胞主要分布在视上核和室旁核，在视上弥散核、弓状核、室周核和乳头体各核团也存在免疫阳性神经元；在室旁核、视上弥散核、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。在下丘脑23个核团（区）中均能检测出OT mRNA的阳性细胞。结果表明，OT和OT mRNA在下丘脑中分布广泛，且OT可能通过轴突传递和血液运输，将OT mRNA合成的OT运送到别的核团；OT在奶山羊发情过程中发挥了重要作用。     

发情周期中奶山羊下丘脑—垂体—卵巢轴催产素的免疫组化定位

用免疫组化ABC法，对发情周期中奶山羊下丘脑—全体—卵巢轴催产素(OT)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室旁核和视上核，在穿窿周核、腹内侧核、腹外侧核、交叉上核、背内侧核、乳头体、下丘脑外侧区、下丘脑前核等核团也有一定数量的阳性神经元；阳性神经纤维仅见于室旁核、下丘脑前核、视上核等少数核团，在正中隆起和第3脑室室周可见到一定数量的阳性神经纤维。在全体前叶未见到OT免疫反应阳性产物，自垂体柄和正中隆起的一例可见到平行排列的OT阳性神经纤维断续地延伸至神经部。卵巢的卵泡及间质未见OT免疫阳性反应，在黄体组织中存在数量较多的免疫反应阳性细胞，阳性细胞主要呈圆形、卵圆形，小梁两侧及黄体中央近腔区域的阳性细胞呈长梭形，有相当数量的阳性细胞具有突起。连续切片HE染色对照观察显示，黄体中OT主要由大黄体细胞产生，但小黄体细胞也存在OT免疫阳性反应。     

羔羊期和性成熟期奶山羊下丘脑中催产素的表达

采用免疫组化链霉素抗生物素蛋白-过氧化物酶法(SP法),检测羔羊期和性成熟期雌性奶山羊下丘脑中催产素表达的变化规律.结果显示,催产素免疫反应阳性神经元在羔羊和性成熟羊的视上核、室旁核、乳头体内侧核等核团均有分布,在羔羊的室周核、视前内侧核等核团广泛分布,但在性成熟羊的这些核团却未见分布.在视上核、室旁核、乳头体内侧核等核团的催产素免疫阳性神经元数量,性成熟羊显著高于羔羊(P＜0.01);这些核团中催产素免疫阳性产物的平均灰度值,羔羊显著低于性成熟羊(P＜0.01),在其他核团则羔羊高于性成熟羊,且大部分差异明显(P＜0.01).结果提示,性成熟羊下丘脑中除室周核、视前内侧核等核团外,室旁核、视上核、乳头体内侧核、乳头体后核等核团,催产素的表达高于羔羊,说明这些核团对生殖系统的发育成熟可能有重要作用.     

Intragastric gavage with denatonium benzoate acutely induces neuronal activation in the solitary tract nucleus via the vagal afferent pathway

Natural toxic substances have a bitter taste and their ingestion sends signals to the brain leading to aversive oral sensations. In the present study, we investigated chronological changes in c-Fos immunoreactivity in the nucleus tractus solitarius (NTS) to study the bitter taste reaction time of neurons in the NTS. Equal volumes (0.5 mL) of denatonium benzoate (DB), a bitter tastant, or its vehicle (distilled water) were administered to rats intragastrically. The rats were sacrificed at 0, 0.5, 1, 2, 4, 8, or 16 h after treatment. In the vehicle-treated group, the number of c-Fos-positive nuclei started to increase 0.5 h after treatment and peaked 2 h after gavage. In contrast, the number of c-Fos-positive nuclei in the DB-treated group significantly increased 1 h after gavage. Thereafter, the number of c-Fos immunoreactive nuclei decreased over time. The number of c-Fos immunoreactive nuclei in the NTS was also increased in a dose-dependent manner 1 h after gavage. Subdiaphragmatic vagotomy significantly decreased DB-induced neuronal activation in the NTS. These results suggest that intragastric DB increases neuronal c-Fos expression in the NTS 1 h after gavage and this effect is mediated by vagal afferent fibers.     

The developmental pattern of c-fos expression in the rat thalamus following open-field stress stimulation

The thalamic nuclei with their defined set of input-output connections are the primary channel for information flow to the cerebral cortex. Several data suggest that neurons of that area are involved in the response to various aversive stimulations. However the pattern of activation seems to depend on the stress model as well as the stage of maturation. In the present study we would like to check which nuclei of the thalamus show expression of c-fos in the response to the "open field test", and how this response pattern changes during the maturation process. 30 rats of age ranged from P0 to P120 (P-postnatal day) were studied. The experimental group was exposed to the "open field test" for 10 minutes. After perfusion and fixation, brains were cut and stained for c-fos with immunohistochemical method. Our results showed that during development the pattern of c-fos activity in the thalamic nuclei after stress stimulation undergoes significant changes. Distinct c-fos expression was observed in the paraventricular nucleus, intergeniculate leaflet and ventral lateral geniculate nucleus. These findings suggest that these nuclei may play a direct role in the stress reaction involved in the response to the "open field test".     

Estrogen receptor alpha expression in the medial preoptic area and the medial basal hypothalamus under different physiological conditions in cattle

The distribution and morphology of immunoreactive estrogen receptor alpha (ERalpha)- containing cells were examined in the cattle brain. The brains were collected from two cows and a male calf. One of the cows was sacrificed one week after parturition. ERalpha expressions in the preoptic area of the rostral forebrain and the medial basal hypothalamic area were evaluated immunohistochemically using a mouse monoclonal antibody. In all animals, the signals that indicate ERalpha were detected in the medial preoptic area, the level of the organum vasculosum of the lamina terminalis and the bed nucleus of the stria terminalis. In the caudal hypothalamic region, they were detected in the arcuate nucleus, the periventricular and ventromedial hypothalamic nuclei. ERalpha-containing cells were characterized by a dense nuclear reaction product. In all the subjects examined, ERalpha signals were clearly detected in the cytoplasm as well. The density of ERalpha expression was different among the three cattle; the number of ERalpha containing cells in the postpartum cow was much lower than that in the other cow and the male calf. The present findings suggest that ERalpha expression in the forebrain is influenced by the reproductive status in the cattle.     

猕猴脾脏中雌激素受体的表达及其性别差异

选取健康成年雌雄猕猴各3只,采用免疫组织化学SP法研究了雌激素受体(ER)在脾脏中的分布,观察猕猴脾脏中ER的表达及性别差异.结果显示,ER免疫阳性反应物主要分布于脾脏红髓区,脾小结相对较少,而动脉周围淋巴鞘、血管内皮、脾小粱等组织结构内仅有少量分布.ER阳性产物主要定位于细胞核中.部分存在于胞浆和胞膜上.雌性猕猴脾脏中的阳性细胞数量显著高于雄性,表达强度也较雄性强.这表明,ER参与雌激素对脾脏的免疫功能调控.ER阳性产物分布特点表明雌激素发挥作用主要是通过经典基因组机制,同时也通过非基因组机制途径.而ER表达的明显性别差异提示体内雌激素水平可能对脾脏中B淋巴细胞的功能有正调节作用.     

Immunohistochemical demonstration of chromogranin A in endocrine organs of the rat and horse by use of region-specific antibodies

Chromogranin A (CgA) is an acidic glycoprotein that is co-stored with hormones or neurotransmitters in granular components of endocrine cells and neurons, and released together with them in response to adequate stimulation. In addition to acting as a packaging protein, CgA functions as a precursor molecule that yields several bioactive peptides by proteolytic cleavage. The purpose of this study is to elucidate how different the processing of CgA is among endocrine tissues by immunostaining using multiple region-specific antisera, and to evaluate the availability of region-specific antisera. When various endocrine organs of rats were immunostained with four region-specific antisera against rat CgA (CgA 1-28, 94-130, 296-314, and 359-389), all amine/peptide-secreting endocrine tissues except the pineal body were stained positively. The adrenal medulla and gastric endocrine cells were equally intensely immunoreactive to all four antisera, while the other endocrine tissues, represented by pancreatic islets, showed different staining patterns depending on the antiserum. These results suggest that the processing of CgA differs from tissue to tissue. An antiserum against horse CgA 335-365, corresponding to rat CgA 359-389 which shows the highest concentration in the plasma and urine of the rat, again stained all endocrine tissues of the horse except the pineal body. Therefore, the anti-horse CgA 335-365 serum is useful for immunohistochemical survey of horse CgA, and may make possible the establishment of a CgA assay system for the measurement of CgA in the plasma, urine and saliva.     

ChAT在1日龄锥鸡丘脑视、听觉中继核团内的分布

胆碱乙酰转移酶（ChAT)是ACh的合成酶，为了观察它在禽脑内的分布，试验选取1日龄雏鸡5天，采用免疫组化SP法研究了ChAT在鸡丘脑视、听觉中继核团内的分。结果表明，在1日龄雏鸡丘脑中，ChAT阳性神经元广泛分布于圆核、卵圆核及亚圆核中，且密度较高、染色较深；外侧膝状体核中既有阳性神经元，又有阳性纤维分布。这表明1日龄雏鸡丘脑中继核团胆碱能神经系统的发育已趋于完善。     

泰和乌鸡脑桥9个神经核团的细胞构筑

为了探明乌鸡中枢神经系统内神经核团的细胞构筑,本试验以8羽泰和乌鸡为研究对象,采用石蜡切片和HE染色技术,在光镜下观察分析乌鸡脑桥内耳蜗神经核、外展神经核及其副核、面神经核、三叉神经感觉主核、三叉神经运动核、蓝斑、蓝斑下核和脑桥中缝核的形态特征。结果表明:乌鸡脑桥的耳蜗神经核也分为耳蜗神经大细胞核、板核、角核3个亚核,核团内神经元以中型为主,耳蜗神经大细胞核相对比较发达,角核核周界不明显;外展神经核吻极与面神经核吻极几乎平行,外展神经副核与外展神经核非常接近,不易分开;面神经中间核不发达;三叉神经运动核没有明显的内、外、腹侧亚核之分;蓝斑细胞排列较密集,细胞多为圆形、椭圆形,以中型细胞为主;蓝斑下核细胞排列松散,核团界限不明显,绝大多数细胞为椭圆形、纺锤形,中型细胞占多数。中缝脑桥核位于脑桥后段中部以下的中缝区,神经元多为三角形、星形和椭圆形,突起很发达,多为巨型细胞。各核团的分布与北京鸭和鸡的大体相似。     

Estrogen receptors in canine mammary gland tumours

The aim of the present study was to establish the relationship between estrogen receptor expression in neoplastic parenchymal cells and the type of canine mammary gland tumour. The research material included mammary gland tumours obtained from 66 dogs. Paraffin sections were stained with hematoxylin-eosin and immunohistochemically with monoclonal antibodies and LSAB/Peroxidase/Universal Kit. The estrogen receptor expression was observed in nuclei of neoplastic cells in about 59% of the cases and in the cytoplasm in about 89% of the cases. In about 20% of the cases the expression in the nuclei and in the cytoplasm was extremely weak. No correlation was found between the expression of estrogen receptors and the value of mitotic indexes in the neoplasms investigated.