大鼠端脑内一氧化氮合酶阳性神经元的发育

目的研究大鼠胚胎时期及生后早期一氧化氮合酶（NOS）阳性神经元在端脑的分布,探讨一氧化氮（NO）在脑发育过程中的作用。方法应用还原型尼克酰胺腺嘌呤二核苷酸磷酸脱氢酶（NADPH-d）组织化学方法观察孕14d起至生后14d大鼠端脑内NOS阳性神经元的形态和分布。结果孕14d没有观察到阳性神经元。孕15d纹状体腹外侧已有NOS阳性表达。孕17d在大脑皮质、梨状皮质观察到NOS阳性神经元,但胞体小,树突短,且分支少。随着年龄的增长神经元的胞体数目增多、染色增强或维持一定的水平。到孕20d,NOS阳性神经元分布广泛,梨状皮质、纹状体腹外侧及终纹床核均有大量NOS阳性神经元,其胞体明显增大,树突分支复杂化,长度增加。在生后,除上述脑区的阳性神经元进一步发育分化,大脑皮质和纹状体的NOS阳性纤维相互编织成疏密不等的纤维网外,在胼胝体、海马也观察到NOS阳性神经元。到生后14d,NOS阳性神经元的分布模式总体上已与成年大鼠相似。结论NOS阳性神经元在端脑独特的表达模式提示NO在脑发育和成熟过程中扮演重要角色。     

大鼠纹状体一氧化氮合酶阳性神经元的生后分布及发育

目的：探讨一氧化氮合酶(NOS)阳性神经元在生后不同日龄(1、5、10、15、30、120d)大鼠纹状体中的分布及形态变化特征。方法：NADPH-d组织化学方法。结果：NOS阳性神经元首先出现于尾壳核前上部，随生长发育逐渐扩展至外侧部、内侧部和苍白球；1～5d时胞体直径10μm左右，无突起，或仅1～2个短小突起，10～15d胞体增大，数量增多，突起明显增多伸长，并有许多膨体，30d时胞体直径达20～35μm，突起200μm以上，并反复分支交织成网状，120d与30d比较无明显变化。结论：纹状体内NOS阳性神经元的发育变化在生后1～30d之间，NO可能与运动机能的建立及调控有关。     

迷走神经切断术后小鼠胰岛内神经元型一氧化氮合酶免疫阳性神经末梢的变化

目的:观察迷走神经切断术后胰岛内神经元型一氧化氮合酶(nNOS)免疫阳性神经末梢分布的变化.方法:将28只小鼠随机分成实验组和对照组,对实验组小鼠行膈下食管迷走神经前、后干切断术,对照组不做任何处理.术后3、 5、 7、 14d分别处死小鼠,取胰冷冻切片,免疫组织化学显色.结果:在实验组和对照组的胰岛内均可观察到nNOS免疫阳性的副交感神经节和神经末梢存在,神经末梢在胰岛周围或深入内部,呈树枝状或念珠状分布,大多成束走行.迷走神经切断后小鼠胰内胰岛nNOS免疫阳性神经末梢的分布密度明显低于正常对照组.结论:胰岛内nNOS免疫阳性神经末梢与迷走神经存在较大的相关性,提示NO是副交感神经调节胰岛分泌的主要神经递质之一.     

应激性防御反应过程中脑内一氧化氮合酶阳性神经元分布的变化

本实验采用 NADPH-d方法研究发现 :在应激的早期 ( 1～ 6d) ,一氧化氮合酶阳性神经元在大脑皮质、基底前脑、纹状体、间脑和脑干内出现的部位增多 ,一氧化氮合酶阳性神经元的数量也增多 ;而在应激的晚期 ( 9d以后 ) ,一氧化氮合酶阳性神经元出现的部位及数量明显减少。提示在慢性应激的早期一氧化氮合酶活性增高 ,合成一氧化氮的能力增强 ;而在应激的晚期一氧化氮合酶活性降低 ,合成一氧化氮的能力降低     

大鼠脑干神经元型一氧化氮合酶免疫阳性神经元的分布

目的　观察大鼠脑干神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的分布 ,为探讨nNOS的作用提供形态学资料。方法　用ABC免疫细胞化学方法显示脑干nNOS免疫阳性神经元。结果　大鼠脑干nNOS免疫阳性神经元以中脑和脑桥分布丰富 ,延髓较稀少 ;在中脑 ,nNOS免疫阳性神经元主要分布于中脑水管周围灰质的背侧部、被盖背外侧核、中缝背核、上下丘灰质等部位 ;在脑桥 ,主要分布于被盖背外侧核、脑桥中缝核、被盖脚桥核、蓝斑、臂旁核、斜方体核 ,以及脑桥网状结构 ;与中脑和脑桥相比 ,延髓nNOS免疫阳性神经元较少 ,主要分布于延髓网状结构、三叉神经脊束核和孤束核等核团。结论　分布于脑干内丰富的nNOS免疫阳性神经元可能通过其生成的NO调节其他神经递质的分泌 ,共同参与内脏活动、感觉和运动的传导 ,以及睡眠和觉醒等脑的高级整合功能的调节。     

杏仁核簇一氧化氮合酶（NOS）阳性神经元的分布

采用NADPH－d组织化学方法，观察了NOS阳性神经元在大鼠杏仁核簇内的分布和形态特征，结果表明，杏仁核簇的大部分核团都含有NOS阳性神经元，其中，杏仁前区，杏仁基底核外侧部，杏仁基底核内侧部，杏仁外侧核后部，杏仁皮质核和杏仁内侧核均观察到较多的NOS阳性神经元，杏仁外侧核前部和杏仁中央核也见到少量NOS阳性神经元。     

胚鼠大脑皮质一氧化氮合酶阳性神经元的体外生长发育研究

本文在体外培养条件下观察了孕 14 d SD胚鼠大脑皮质中一氧化氮合酶阳性神经元的形态和发育过程。将孕 14 d SD胚鼠大脑皮质细胞悬液接种于 2 4孔培养板 ,实验按培养龄分 4组 ,分别在接种后 5d、10 d、15d和 2 0 d终止培养 ,继而进行NA DPH - d组织化学染色 ,观察各组一氧化氮合酶阳性神经元的数量和形态。结果显示 :培养后 5d即有一定数量的一氧化氮合酶阳性神经元出现 ,在 4个实验组中 ,以 15d组的一氧化氮合酶阳性神经元的数量为最多、突起最长 ,深染细胞数量也最多 ,大部分一氧化氮合酶阳性神经元的胞体与突起均局限分布在细胞团块内 ,胞体大小不一 ,只有少量的一氧化氮合酶阳性细胞散在分布在细胞团块之间 ,其突起伸向或围绕团块。本研究结果表明 ,在胚鼠大脑皮质的神经元内即已有一氧化氮合酶存在 ,提示一氧化氮可能与胚脑的生长发育有关     

8-精加压素免疫反应阳性神经元在兔脑内的分布

用纯种新西兰兔,向侧脑室内注入秋水仙素(300μg/3μl),48h后用免疫组织化学方法观察了加压素(VP)免疫反应阳性神经元在兔脑内的分布。实验结果表明,在室旁核有较密集的VP样免疫反应阳性神经元分布,胞体直径约为15—20μm。在下丘脑另一些部分、视前区、杏仁核和蓝斑均有VP样免疫反应阳性的神经元,但细胞不如室旁核的密集,在蓝斑的阳性细胞则较小。另外,室周灰质和臂旁核也发现有少量VP样免疫反应阳性的细胞。本工作还首次报道了在蓝斑的小血管壁上有VP样免疫反应阳性神经末梢分布,推测这些神经末梢可能参与VP调节血压的中枢机制。     

一氧化氮合酶阳性神经元及纤维在大鼠脊髓的分布及其来源

用还原型尼克酰胺腺嘌呤二核苷磷酸(NADPH)脱氢酶反应观察了大鼠脊髓内一氧化氮合酶阳性神经元和纤维的分布。胞体浓染且突起显示良好的类似Golgi染色的阳性神经元分布于亘脊髓全长的中央管周围灰质(X层),胸腰髓(T1-L3)的中间带外侧核、中介核及腰骶髓(L6、S1)的骶髓副交感核和后连合核,少量弥散于后角Ⅳ～Ⅵ层。后角Ⅲ层内可见较多的胞体浓染或淡染但突起短或未见阳性突起的小型神经元。前角内无阳性神经元,但有较多串珠状阳性终末支,并在运动神经元胞体和树突上形成终扣。阳性纤维和终末在后角Ⅰ层和Ⅲ层密集,其它部位稀少。在胸腰段和腰骶段脊髓后角内、外侧缘有行向腹侧的内、外侧阳性纤维束,外侧束集中而粗大,阳性纤维束终止于中间带外侧核和后连合核。切断后根和半横断颈髓的实验表明,后角Ⅰ层的阳性纤维和终末主要来源于脊神经节内一氧化氮合酶阳性细胞的中枢突,而Ⅲ层内的阳性纤维和终末则为Ⅲ层和后角深层阳性细胞的突起。结合文献和本文结果可以推断,一氧化氮在躯体和内脏感觉的传入过程及内脏传出活动中是一种重要的活性物质。     

小鼠心脏一氧化氮合酶神经纤维的分布

目的：探讨小鼠心脏一氧化氮合酶aninic oxide synthse神经元结构和神经纤维分布。方法：NADPH-d组织化学技术。结果：小鼠心脏神经元主要分为NOS强阳性、中度和弱阳性反应细胞；心脏各部均接受NOS神经支配，其神经纤维多与肌纤维长轴平行走向。心房最丰富，房室结次之，左、右心室最少。心房及房室结的NOS阳性纤维呈串珠状膨大，心室的常为丝状，膨体极少。结论：小鼠心脏NOS神经元包括强阳性、中度和弱阳性反应细胞，NOS阳性纤维在心脏各部的分布和形态均有差异，NO可能作为神经递质和/或神经调节剂在心血流和冲动传导等的神经调控中起作用。     

Spatial organization and morphometric characteristics of histaminergic neurons in the rat brain

We report here a study addressing the spatial organization, densities, numbers, sizes, and shapes of histaminergic neurons in the rat hypothalamus. Studies were performed on 50 rats using histochemical and morphometric methods, computer image analysis, and three-dimensional reconstruction of the histaminergic nuclei of the hypothalamus. The results demonstrated that the total bilateral volume of the histaminergic nuclei of the rat brain amounts to 0.5 mm³: the E2 nucleus occupies 40% of the total volume, E4 occupies 35%, E3 occupies 13%, E5 occupies 9%, and E1 occupies 3%. The distribution density of monoamine oxidase B neurons in the histaminergic nuclei of the rat hypothalamus decreased in the order E1 > E3 > E2 (the “compact” nuclei) > E4 (an “intermediate” density nucleus) > E5 (the “diffuse part”). The mean number of histaminergic neurons in the rat hypothalamus was 37200 ± 2800; the E2 nucleus contained 54% of these cells, E3 contained 23%, E4 contained 6%, E1 contained 7%, and E5 contained 0.32%. The E1–E3 nuclei were dominated by small and intermediate-sized neurons, round in shape, and the E5 nucleus was dominated by intermediate and large neurons, fusiform in shape. A subpopulation of giant histaminergic neurons was detected in the E4–E5 nuclei. __________ Translated from Morfologiya, Vol. 127, No. 2, pp. 27–30, March–April, 2005.     

Metabolic changes in rat brain histaminergic neurons during subhepatic cholestasis

The aim of the present work was to assess metabolic changes in histaminergic neurons in the rat brain during subhepatic cholestasis. Studies were performed on male Wistar rats using quantitative histochemical methods. The results showed that in cholestasis, histaminergic neurons in the rat hypothalamus developed significant changes in succinate dehydrogenase, lactate dehydrogenase, and glucose-6-phosphate dehydrogenase activity, in NADH and NADPH, and in acid phosphatase and monoamine oxidase B. These changes depended on the duration of cholestasis and had a dynamic, wave-like nature. The changes were apparent after five days of cholestasis, reached a maximum at 10–20 days, decreased at 45 days, and completely disappeared at 90 days. __________ Translated from Morfologiya, Vol. 132, No. 4, pp. 27–30, July–August, 2007.     

Further characterization of sleep-active neuronal nitric oxide synthase neurons in the mouse brain

We recently demonstrated that Fos is induced in a subpopulation of cortical neuronal nitric oxide synthase (nNOS)-immunoreactive neurons in three rodent species both during spontaneous sleep (SS) and recovery sleep (RS) after a period of sleep deprivation (SD); the proportion of cortical Fos⁺/nNOS neurons was significantly correlated with non-REM (NREM) sleep delta energy. The present study was undertaken to evaluate the specificity of this state-dependent activation of cortical nNOS cells. The percentage of nNOS neurons that expressed Fos during SD and RS was determined in nine subcortical brain regions and the cortex of the mouse brain; a significantly greater proportion of Fos⁺/nNOS neurons was observed during RS only in the cortex and in none of the nine subcortical regions. The proportion of calretinin-, calbindin- and parvalbumin-immunoreactive cortical interneurons that expressed Fos during SD and RS was also determined. In contrast to cortical nNOS neurons, a higher percentage of Fos⁺/calbindin neurons was found during SD than RS; there were no differences in the proportions of Fos-expressing parvalbumin or calretinin neurons between these conditions. Since the nNOS and calretinin cortical interneuron populations overlap extensively in the mouse brain, triple-labeling with these two phenotypic markers and Fos was undertaken in mice from the RS group to determine which combination of markers could best identify the rare “sleep-active” cortical interneuron population. The proportions of both Fos⁺/nNOS neurons and Fos⁺/nNOS/calretinin neurons far exceeded the proportion of Fos⁺/calretinin neurons during RS, but the proportions of these two cell types were not significantly different during RS. Thus, functional activation of nNOS neurons during sleep appears to be restricted to the cerebral cortex and cortical nNOS cells and nNOS/calretinin cells collectively define a cortical interneuron population that is activated during sleep.     

扬子鳄胸髓一氧化氮合酶和乙酰胆碱酯酶阳性神经元的分布

目的观察一氧化氮合酶(NOS)和乙酰胆碱酯酶(AChE)阳性神经元在扬子鳄胸髓的分布。方法采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)法和亚铁氰化铜法观察扬子鳄胸髓NOS和AChE阳性神经元的分布。结果扬子鳄胸髓前角、后角和中央灰质内可见NOS和AChE阳性神经元,白质内含有丰富的NOS和AChE阳性神经纤维。结论扬子鳄胸髓有NOS和AChE阳性神经元分布。     

Expression of nitric oxide synthase and effect of substrate manipulation of the nitric oxide pathway in mouse ovarian follicles

BACKGROUND: Nitric oxide (NO) is a cell messenger with multiple actions in different biological systems, implicated in the control of follicle and oocyte function. NO is formed from L-arginine by isoforms of nitric oxide synthase (NOS) via NG-hydroxy-L-arginine, with L-citrulline as a byproduct. This study aimed to show how modulation of NO by manipulating NOS substrates would affect mouse follicle growth and ovulation in vitro, where vascular effects of NO are attenuated. METHODS: Immunohistochemistry [endothelial (eNOS) and inducible (iNOS)] and in situ hybridization (iNOS) were applied on mouse ovaries. Cultured follicles were also stained for iNOS by immunohistochemistry. For follicles cultured in the presence or absence of L-arginine, the ability of L-citrulline or NG-hydroxy-L-arginine to substitute for L-arginine was assessed in terms of follicle growth and ovulation. RESULTS: iNOS and eNOS were localized in oocytes and theca, with some staining in granulosa. iNOS mRNA occurred predominantly in granulosa and oocyte. Omission of L-arginine significantly reduced follicle survival and ovulation. Partial compensation for L-arginine withdrawal was achieved with L-citrulline and NG-hydroxy-L- arginine. Specific abnormalities of follicle growth were noted. CONCLUSIONS: NOS is present in mouse follicles, and its action is necessary at a local level for normal follicle development in vitro. Reduced growth, persistent basement membranes and reduced ovulation were associated with in vitro disruption of NO.     

Structural-metabolic changes in histaminergic neurons of the rat hypothalamus in conditions of loss of bile

The aim of the present work was to evaluate structural and metabolic changes in histaminergic neurons in hypothalamic nucleus E2 in rats in conditions of complete external drainage of bile. Studies were performed on male Wistar rats (n = 45). Controls consisted of animals subjected to sham surgery with preservation of physiological bile flow throughout the experiment. Quantitative histological and histochemical methods were used. Serial frontal cryostat sections cut from the posterior hypothalamus were used for detection of the activity of the following enzymes: monoamine oxidase B, succinate dehydrogenase, NADH dehydrogenase, NADPH dehydrogenase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, and acid phosphatase. Morphological studies of histaminergic neurons were performed on preparations stained with thionine. These studies showed that complete external drainage of bile led to transient size reductions and rounding of cell perikarya. Metabolic changes were seen within a day of bile loss and subsequently progressed. All energy metabolic pathways were suppressed and acid phosphatase activity was increased on day 5. __________ Translated from Morfologiya, Vol. 132, No. 6, pp. 22–25, November–December, 2007.     

应激性高血压大鼠中脑导水管周围灰质内一氧化氮合酶阳性神经元的变化

目的 :研究应激性高血压大鼠中脑导水管周围灰质 (periaqueductalgray ,PAG)内一氧化氮合酶 (nitricoxidesynthase,NOS)阳性神经元的变化在该病发生中的作用。方法 :采用电击足底结合噪声建立应激性高血压大鼠模型 ,NADPH d和免疫组化方法观察PAG内NOS阳性神经元的变化 ,并进行光镜计数和图像分析。结果 :应激性高血压大鼠PAG内NOS阳性神经元和神经元型 (neuronalNOSnNOS)免疫阳性神经元的数量均明显减少 ,平均灰度值明显增高 ;且二者的数量和平均灰度值与应激性高血压大鼠的收缩压均有明显相关关系。结论 :应激性高血压大鼠PAG内NOS阳性神经元的变化可能参与了应激性高血压的形成。