神经生长因子受体和一氧化氮合酶在大鼠基底前脑神经元的分布与共存

用神经生长因子受体免疫组化，NADPH－d组化及两者的双重反应技术观察了神经生长因子受体和一氧化氮合酶在大鼠基底前脑神经元的表达和共存。神经生长因子受体和一氧化氮合酶阳性神经元在基底前脑内都以一条连续的细胞带的形式存在．神经生长因子受体阳性神经元多为多极形．排列较密，由吻侧向尾侧数量逐渐增多；一氧化氮合酶神经元的形状与神经生长因子受体阳性神经元相似，但数量较少，且由吻侧向尾侧数量相对稳定。通过双重反应证明，基底前脑神经元包括着神经生长因子受体阳性和一氧化氮合酶阳性以及两者共存的神经元等三种类型。双重反应阳性神经元在形态上和神经生长因子受体阳性神经元相似，其数量从吻侧向尾侧逐渐减少，在前由＋1．0水平，它占一氧化氮合酶阳性神经元的70％，神经生长因子受体阳性神经元的50％；但到前连合水平则仅占一氧化氮合酶阳性神经元的20％和神经生长因子受体阳性神经元的40％。本文对神经生长因子受体与一氧化氮合酶双重反应阳性神经元的分布意义以及此二种阳性神经元之间的关系进行了讨论。     

Exposure to aluminium changes the NADPH-diaphorase/NPY pattern in the rat cerebral cortex

Aluminium (Al) impairs the glutamate-nitric oxide-cGMP pathway and reduces the number of nitroxidergic neurons in the rat somatosensory cortex. To understand better the effect of the time of exposure, we monitored the effect of aluminium administration on nitroxidergic neurons, identified by NADPH-diaphorase (NADPH-d) or by nitric oxide synthase (NOS) staining, after 0.5, 1, 2, 3, 6 and 12 months of aluminium administration. Since neuropeptide Y (NPY) is known to be colocalised with nitric oxide synthase in cortical neurons, the aim of this work was to study the effects of Al administration on the cortical expression of NADPH-d, nNOS, and NPY. NADPH-d or NOS positive neurons were found scattered in the cortex where they constituted about 1% of all neurons. Double staining using NADPH-d and NPY showed that almost all nitroxidergic neurons were co-localised with NPY neurons (NADPH-d/NPY double stained neurons) whereas some neurons were stained only with NPY (NPY single stained neurons) ; these were more numerous than NADPH-d/NPY double stained neurons. Al significantly reduced NADPH-d and nNOS positive neurons in the cerebral cortex time dependently, with the greatest effect appearing after 3 months. Also measured was the integrated optical density (IOD) of nNOS positive neurons showing a significant decrease of NOS immunostaining even in the remaining NOS positive neurons. The double staining experiment exhibited a decrease in NADPH-d/NPY double stained neurons with an apparent increase in NPY single stained neurons; these then decreased after 6-12 months. On the whole, the results confirm that Al impairs nitroxidergic pathways time dependently; moreover, the transient increase in NPY single stained neurons from 1 to 3 months suggests that there is an intraneuronal down-regulation of NOS, without affecting neuronal viability. In addition, the decrease in the NPY system found at 6 and 12 months may indicate that Al affected nitroxidergic and NPY systems at different times.     

Changes in mRNA of protein inhibitor of neuronal nitric oxide synthase following facial nerve transection

Protein inhibitor of neuronal nitric oxide synthase (PIN) is reported as the protein inhibiting neuronal nitric oxide synthase (nNOS) activity by preventing dimerization of nNOS. It was also reported that PIN inhibits the activity of all nitric oxide synthase (NOS) isozymes. We examined the effects of facial nerve transection on PIN mRNA and NOS expression by in situ hybridization for PIN mRNA and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) staining. PIN mRNA was initially expressed and transiently increased from 3 to 5 days and returned to the basal level at 7 days after axotomy in the motoneurons of the facial nucleus. NADPH-d-positive motoneurons were found from 7 days post-operation in the facial nucleus. These results suggest that PIN may interact with NOS from 7 days post-operation.     

成人基底前脑巢蛋白免疫阳性神经细胞的细胞化学观察

目的 探讨巢蛋白(nestin)免疫阳性细胞在人基底前脑内的分布规律及化学特性。方法 采用nestin331B、10C2、Rat 401抗体显示人基底前脑的nestin免疫阳性细胞，并应用NSE、p75NGFR、ChAT、GFAP抗体分别对nestin免疫阳性细胞进行了双标免疫细胞化学研究，同时还应用NADPH-d进行组织化学染色。结果 在成人基底前脑有一个连续的nestin免疫阳性细胞带，胞体较大，呈卵圆形或梭形，有1～3个突起，分布于隔核、斜角带核、Meynert核、无名质及杏仁核群。双标染色显示，nestin阳性细胞被NSE抗体标记，并与ChAT、NGFR、NOS阳性神经元间杂分布，多数不呈交叉反应。约28％nestin免疫阳性细胞为ChAT阳性神经元，15％为NGFR阳性神经元，6％为NOS阳性神经元。此外，透明隔及隔区靠近软膜处有少量nestin免疫阳性细胞，其形态与星形胶质细胞相似，不与GFAP有交叉免疫阳性反应。结论 成人基底前脑存在一个有别于ChAT、NGFR、NOS神经元的nestin免疫阳性神经元簇，其生物学意义值得进一步研究。     

Direct projections from the extrathalamic forebrain structures to the neocortex in the macaque monkey

Extrathalamic direct projections from the subcortical forebrain structures to the neocortex were examined in the macaque monkey by the horseradish peroxidase method. The enzyme, when injected into discrete regions in the neocortex, labeled cell bodies of extrathalamic forebrain neurons in the basal nucleus of Meynert, nucleus of the diagonal band, medial septal nucleus, hypothalamus, claustrum and dorsolateral part of the basal amygdaloid nucleus. Neurons in the basal nucleus of Meynert, lateral hypothalamus and claustrum appeared to send their axons widely, but not diffusely, to the neocortex.     

A cytoarchitectonic and histochemical study of nucleus basalis and associated cell groups in the normal human brain

Several recent studies have reported loss of neurons in the nucleus basalis in Alzheimer's disease. However, few detailed studies of the normal distribution of these neurons in the human brain have appeared. We have used Nissl staining and acetylcholinesterase histochemical staining of the human basal forebrain, alone or in combination to identify the organization of the nucleus basalis and associated cell groups, (or collectively, the magnocellular basal nucleus) in the normal human brain. The magnocellular basal nucleus includes a series of clusters of neurons and scattered perikarya extending from the medial septum and diagonal band nucleus rostrally, through the substantia innominata to the furthest caudal extent of the globus pallidus. This distribution is similar to that which has been described in the monkey. Furthermore, acetylcholinesterase-positive fibers in the human brain are seen in the two major pathways that have been identified as carrying magnocellular basal nucleus axons to the cerebral cortex in other species.These observations suggest that the topographic organization of the magnocellular basal projection to cerebral cortex in other species probably exists in man as well. It will therefore be important in future studies of the fate of these neurons in neurological degenerative diseases to assess the loss of neurons in the different components of the magnocellular basal nucleus in relation to the clinical evidence for dysfunction in the cortical areas which they innervate.     

成年大鼠基底前脑存在一个Nestin免疫阳性神经元簇

目的　观察Nestin免疫活性在成年大鼠基底前脑中的表达和分布 ,并探讨其与胆碱能神经元之间的关系。方法　采用免疫组化方法对成年大鼠基底前脑的切片进行nestin免疫组化染色及其与ChAT ,NADPH d双标染色。结果　在成年大鼠基底前脑的隔斜角带复合体有一个连续的nestin免疫阳性细胞带 ,胞体较大 ,梭形或多极形 ,有 2～ 4个突起。双标染色显示 ,Nestin阳性神经元与ChAT ,NADPH d阳性神经元间杂分布 ,大多数不呈交叉反应 ,只有少数 ,约 10 %呈双标染色。结论　成年大鼠基底前脑存在一个有别于胆碱能神经元的nestin免疫阳性神经元簇 ,其化学属性和生物学意义有待进一步研究。     

Subcellular localization of neuronal nitric oxide synthase in the superficial gray layer of the rat superior colliculus

The superficial layers of the rat superior colliculus (sSC) receive innervation from retina and include nitric oxide synthase (NOS)-immunoreactive neurons. We used electron microscopic immunocytochemistry to assess the subcellular localization of neuronal NOS (nNOS) in the sSC. nNOS immunoreactivity was detected on the external membrane of mitochondria, endoplasmic reticulum, in pre- and postsynaptic profiles and also diffusely distributed in the cytosol. Postsynaptic labeled regions were often associated with presumptive retinal unlabeled terminals. Microtubules also appeared intensely labeled. These results show that NOS immunoreactive neurons may be innervated by retinal terminals and suggest an association of nNOS with cytoskeletal elements.     

急性缺氧小鼠海马CAl区NOS活力和nNOS蛋白表达的时程变化

目的:观察急性缺氧小鼠海马CAl区一氧化氮合酶(NOS)和神经元型一氧化氮合酶(nNOS) 阳性神经元的时程变化,探讨NO在脑缺氧中的作用并为抗脑缺氧提供依据。方法:复制小鼠急性缺氧模型,采用NADPH-d组织化学和nNOS免疫组织化学方法,研究急性缺氧后不同时程点小鼠海马CAl区NADPH-d 和nNOS阳性神经元数量的变化。结果:与正常对照组相比较,急性缺氧后0.5h组小鼠海马CAl区NADPH-d 和nNOS阳性神经元的数量无明显变化,差异无显著性(P>0.05),3h、6h和12h组逐渐增多并于12h升高达到最高峰,差异有显著性(P<0.05),而于24h后开始降低,48h恢复正常。结论:急性缺氧后早期海马CAl区NOS和nNOS水平明显增多,NO在缺氧所致早期脑损伤中起重要作用。     

Non-cholinergic basal forebrain neurons project to the contralateral basal forebrain in the rat

Following injections of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) or the fluorescent tracer fluoro-gold into the magnocellular preoptic area and the horizontal limb of the diagonal band, retrogradely labelled neurons were found in the homotopic region of the contralateral basal forebrain. Labelled fibers apparently arising from these neurons travelled in the stria medullaris and the habenular commissure to terminate in the contralateral basal forebrain. Although the neurons retrogradely labelled with fluoro-gold in the contralateral basal forebrain were similar in size to choline acetyltransferase (ChAT)-immunoreactive neurons, and were intermingled with them, none was ChAT-positive. WGA-HRP injections into the nucleus basalis magnocellularis did not result in retrograde labelling in the contralateral basal forebrain. These findings suggest that non-cholinergic neurons may serve as a direct link between the two sides of selective magnocellular basal forebrain regions.     

Stability of numbers but not size of mouse forebrain cholinergic neurons to 53 months

In normal mammalian aging there is a reduction of cholinergic markers in a variety of regions. To determine whether this reduction is related to reduced numbers of basal forebrain cholinergic neurons, we counted the number and measured the sizes of the magnocellular acetylcholinesterase-positive neurons in this region of 7, 15, and 53-month-old C57B1/6NNIA mice. Data were collected from coded slides containing the medial septum, nucleus of the diagonal band, magnocellular preoptic nucleus, and nucleus basalis magnocellularis. There was no decline in numbers of basal forebrain acetylcholinesterasepositive neurons in any of the regions studied. However, cell sizes showed a progressive age-related decline which was greatest in the nucleus basalis magnocellularis.     

雄激素替代对去睾丸大鼠基底前脑NOS及Nestin阳性神经元的影响

目的　观察雄激素替代对去睾丸大鼠基底前脑一氧化氮合酶 (NOS)及巢蛋白 (Nestin)阳性神经元的影响。方法　将 2 8只健康成年雄性SD大鼠随机分为 4组 :去势 2 4h雄激素替代组、去势 2w雄激素替代组、去势 2 4h芝麻油替代组及假手术组。用组织化学及免疫组织化学染色方法观察基底前脑NOS和Nestin阳性神经元变化。结果　去势芝麻油替代组内侧隔核 (MS)、斜角带垂直支 (vDB)NOS阳性神经元数明显多于假手术组 (P <0 .0 5 ) ;去势 2 4h或 2w行雄激素替代MS和vDBNOS阳性神经元数少于假手术组水平 (P <0 .0 5、P <0 .0 1) ,但去势芝麻油替代或雄激素替代对斜角带水平支 (hDB)的NOS阳性神经元数影响不大 (P >0 .0 5 )。去势芝麻油替代或雄激素替代 ,Nestin阳性神经数的变化趋势与NOS阳性神经元的相似。结论　去睾丸行雄激素替代 ,可选择性地使基底前脑的NOS、Nestin阳性神经元数明显下降 ,但与替代的时程关系不大 ,提示雄激素可能通过其受体下调NOS和Nestin的表达。     

NOS在大鼠杏仁皮质核传入神经元内的分布

目的研究一氧化氮合酶(NOS)在杏仁皮质核传入投射神经元中的分布。方法采用霍乱毒素B亚单位(CTb)逆行追踪和还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)组织化学双重染色相结合的方法。结果双标神经元(NADPH/CTb)主要分布在孤束核(Sol)、蓝斑(LC)、臂旁内侧核(MPB)、中缝背核(DR)、中央灰质外侧部(CGL)、中央灰质背侧部(CGD)、丘脑室旁核(PV)、下丘脑室旁核(Pa)、下丘脑室周核(Pe)、下丘脑腹内侧(VMH)核以及杏仁内侧核(ME)等神经核团。结论NOS在大鼠杏仁皮质核传入投射神经元中主要分布于上述核团,并且提示NO参与杏仁皮质核的功能调节。     

大鼠斜角带核一氧化氮合酶阳性神经元向嗅球的投射

采用还原型尼克酰胺嘌呤二核苷酸脱氢酶（ＮＡＤＰＨｄ）与辣根过氧化物酶（ＨＲＰ）相结合方法,观察斜角带核一氧化氮合酶（ＮＯＳ）阳性神经元向嗅球的投射。结果发现,当ＨＲＰ注入嗅球后,ＨＲＰＮＯＳ双标神经元出现于斜角带核水平支外侧部和斜角带核垂直支腹侧部,也偶见于斜角带核垂直支背侧部,表明斜角带核有一些ＮＯＳ阳性神经元投射到嗅球     

A double immunocytochemical and histochemical technique for demonstration of cholinergic neurons and microglial cells in basal forebrain and neostriatum of the rat

Cholinergic neurons, and their relationship to microglial cells, have been examined by light and electron microscopy with an immunocytochemical and histochemical double-staining technique, in medial septum, diagonal band of Broca, nucleus basalis and neostriatum. This technique may be applied to experimental models of anterograde and retrograde degeneration of the cholinergic neurons of the basal forebrain in order to collect further information relative to the aetiology of degenerative diseases such as Alzheimer's disease.     

成年大鼠基底前脑隔-斜角带复合体的巢蛋白免疫阳性神经元至海马的纤维投射

目的 观察大鼠基底前脑巢蛋白(nestin)免疫阳性神经元的纤维投射。方法 先用荧光素逆行追踪法显示基底前脑内投射至海马的荧光素标记神经元，观察摄片后再进行nestin免疫组织化学染色。对比荧光照片和免疫组织化学染色照片，辨认荧光素和nestin双标神经元。结果 在基底前脑注射侧的内侧隔核(MS)和斜角带核的垂直支(vDB)和水平支(hDB)均存在荧光素标记细胞，其中一部分为nestin免疫阳性。在MS、vDB和hDB，双标细胞占逆行标记细胞的比例分别为21．3％、25％和20．6％；占nestin阳性细胞的比例分别为26．6％、15．7％和16．3％。结论 大鼠基底前脑的nestin免疫阳性神经元发出神经纤维向海马投射。提示在基底前脑有一个平行于隔-海马胆碱能投射和GABA能投射的第3条通路。     

Cholinergic neurons expressing neuromedin K receptor (NK3) in the basal forebrain of the rat: a double immunofluorescence study

By using a double immunofluorescence method we have examined the distribution of cholinergic neurons expressing neuromedin K receptor (NK3) in the rat brain and spinal cord. The distribution of neuromedin K receptor-like immunoreactive neurons completely overlapped with that of choline acetyltransferase-positive neurons in certain regions of the basal forebrain, e.g. the medial septal nucleus, nucleus of the diagonal band of Broca, magnocellular preoptic nucleus and substantia innominata. Partially overlapping distributions of neuromedin K receptor-like immunoreactive and choline acetyltransferase-positive neurons were found in the basal nucleus of Meynert, globus pallidus, ventral pallidum of the forebrain, tegmental nuclei of the pons and dorsal motor nucleus of the vagus. Neurons showing both neuromedin K receptor-like and choline acetyltransferase immunoreactivities, however, were found predominantly in the medial septal nucleus, nucleus of the diagonal band of Broca and magnocellular preoptic nucleus of the basal forebrain: 66-80% of these choline acetyltransferase-positive neurons displayed neuromedin K receptor-like immunoreactivity. Neurons showing both neuromedin K receptor-like and choline acetyltransferase immunoreactivities were hardly detected in other aforementioned regions of the forebrain, brainstem and spinal cord. The present study has provided morphological evidence for direct physiological modulation or regulation of cholinergic neurons by tachykinins through the neuromedin K receptor in the basal forebrain of rats.     

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。     

Localization of nerve growth factor receptors in the normal human brain and in Alzheimer's disease

NGF receptors were visualized in human brain sections with an immunohistochemical procedure using a monoclonal antibody. This method results in the selective visualization of a population of neurons in the medial septal nucleus, the nucleus of the diagonal band of Broca, and the nucleus basalis of Meynert. Several lines of evidence indicate that this neuronal population is identical to the cholinergic neurons of the basal forebrain. NGF receptor immunohistochemistry therefore represents a sensitive and reliable procedure to selectively visualize forebrain cholinergic neurons for post-mortem analysis. NGF receptors were found to be expressed during the entire life span. However, the intracellular staining intensity was reduced in normal aging, suggesting the tentative conclusion that NGF receptor synthesis may decline in the aged brain. In Alzheimer's disease, the number of NGF receptor-positive cells was decreased. The morphological characteristics of surviving neurons were similar to immuno-positive neurons visualized in normal aged brains.