新生大鼠海马、隔和下丘脑培养细胞中AchE阳性神经元的分布

应用乙酰胆碱酯酶(AchE)组化方法,观察了AchE阳性神经元在新生大鼠海马、隔和下丘脑培养细胞中的分布及形态特征.AchE阳性神经元胞体都较大.在海马培养细胞中AchE阳性神经元数较少,为多极神经元,胞体着色浅.但在隔和下丘脑培养细胞中AchE阳性神经元数较多,多为双极神经元,胞体呈中强度染色.这三种培养细胞中的AchE阳性神经元数均随着培养时间的延长而逐渐增多.     

豚鼠脑干内侧部踏步相关区域AchE阳性神经元的分布

本研究采用胆碱酯酶(AchE)组织化学方法观察豚鼠脑干内侧部踏步相关区域[三叉神经中脑核(Me5)、蓝斑(LC)、中缝大核(RM)及邻近网状结构和延髓巨细胞网状核(Gi)]AchE阳性神经元分布,并利用大鼠和猫进行对照。结果表明豚鼠Me5含有AchE强阳性神经元,而在大鼠和猫的Me5则没有观察到。豚鼠LCAchE阳性神经元分布明显较猫LC的弥散分布集中,虽然密集程度较大鼠LC为弱。此外,在上述三种动物中AchE阳性神经元在Gi、RM及其邻近网状结构具有相似的分布。本文就实验结果与踏步运动的关系进行了探讨。     

家兔喉返神经运动神经元的位置及其性质

本文用辣根过氧化物酶(HRP)、胆碱脂酶(AchE)和脑啡呔(ENK)免疫组化法研究家免喉返神经运动神经元在中枢的位置及其性质。发现:HRP标记细胞主要分布在注射侧的疑核、迷走神经背核。在C_(1,2)节段的背内侧核、面后核以及C_1节段的副神经脊髓核内有少量标记细胞。疑核、迷走神经背核和背内核中有少量AchE—HRP双标记神经元,疑核和背内侧核内可见到ENK—HRP双阳性神经元。这表明家兔喉返神经运动神经元有些是胆碱能神经元和ENK样免疫组化反应阳性神经元。本文对喉返神经运动纤维的周围分布及机能意义进行了讨论。     

乙酰胆碱酯酶阳性神经元在鼠脑的分布和形态特征

本文用乙酰胆碱酯酶(AChE)再生技术,研究AChE阳性神经元在鼠脑的分布和形态特征。按其染色程度,可见强度、中度和轻度三种染色细胞。强染色细胞多数是较大的多极细胞,主要分布于纹状体、基底前脑、下丘脑、黑质、红核、蓝斑,腹侧被盖区、臂旁核、桥被盖核和脑神经运动核。本文将AChE染色结果和胆碱乙酰转移酶(ChAT)免疫组织化学资料进行了比较,对AChE和胆碱能神经元的关系,以及AChE阳性神经元的性质和意义,进行了讨论。     

大鼠脑干臂旁和网状系中AchE阳性结构的分布

采用改良的AchE组化法研究了大鼠臂旁和网状系内AchE阳性结构分布.结果表明:臂旁系的脚桥被蓋核、外背侧被蓋核、臂旁内、外侧核等,均含有AchE阳性细胞和纤维.网状系的脑桥吻、尾侧网状核和延髓巨细胞网状核有散在的AchE阳性细胞和纤维.延髓线形核有排列致密的AchE阳性细胞,系首次报道.     

AchE阳性神经元在中枢的分布、形态特征及海马胆硷能传入的起源——AchE药理组化技术结合HRP追踪

中枢胆硷能机制的研究是当前神经生物学领域活跃的课题,尤其是它在学习、记忆以及某些疾病机制中的作用受到了广泛的重视。由于缺乏理想的显示方法,有关中枢胆硷能神经元的形态学情况了解甚少。现有资料大多来自乙酰胆硷酯酶(AchE)的组织化学方法研究,而脑内大多数区域往往由于染色的神经纤维、神经始末、神经胶质和神经细胞混淆在一起,使观察困难,结果不易评定。本研究以克服上述缺点的AchE药理组化技术(也称 AchE再生技术)系统地研究AchE阳性神经元在鼠、猫脑的分布和形态特征并结合HRP逆行追踪观察海马胆硷能传入的起源。     

DARPP-32在大鼠全脑的定位分布

目的 探讨DARPP-32在大鼠全脑的表达分布特点.方法 应用免疫组织化学染色技术对大鼠脑内DARPP-32的表达分布进行观察.结果 免疫组织化学染色结果显示,强阳性的DARPP-32染色大部分分布于基底节区和前嗅皮质区,主要分布在伏隔核、尾壳核及杏仁核复合体的神经元胞体内,以及苍白球、腹侧苍白球、脚间核及黑质网状部的...     

面神经核内塑形变化在面瘫后遗症发生中的作用

目的观察外伤性面瘫后,面神经核内运动神经元的定位分布与轴索再生变化,探讨其在面瘫后遗症联带发生中的作用。方法大鼠32只,面神经断伤吻合建立面瘫模型,采用尼氏染色与神经逆行荧光示踪技术,观察面瘫60d后健患两侧面神经核内运动神经元的定位分布与轴索再生变化,用透射电镜技术观察面瘫后90d时面神经核内运动神经元的超微结构改变。结果1.面瘫后健患两侧尼氏染色神经元的数量及分布特征基本一致,仅有神经元分布区域的缩小趋势,尼氏染色与超微观察均未发现面神经核内运动神经元有坏死与凋亡样改变。但患侧代表轴索成功再生的荧光金(FG)标记神经元数目较健侧明显减少(P<0.05),提示核内存在未被FG逆行标记但被尼氏染色的未达靶神经元。2.面神经颞支与颊支的荧光红(FR)与FG逆行示踪发现,面神经核颊支支配区内错位分布支配颞支的神经元,患侧的错位神经元比例较健侧增加(P<0.05)。同时,患侧面神经核内还出现代表轴索芽生,同时支配颞颊支的荧光双标神经元。结论外伤性面瘫后面神经核内的塑形中,除轴索芽生与迷向再生影响瘫后功能恢复外,核内还存在未成功再生的面运动神经元,其在面瘫后遗症联带发生中的作用也值得深入研究。     

P19细胞向神经元诱导分化的实验研究

目的 探讨P19细胞向神经元定向分化发育的可能性。方法 经维甲酸(RA)诱导4d的．P19细胞按神经细胞培养方法培养12d，分别用NSE、MAP-2和TH进行免疫组织化学染色，用AchE进行组织化学染色，鉴定分化细胞的特征。结果 经RA诱导的P19细胞培养12d，大多数细胞转变为类似于神经元的形态，长出突起并相互联络成网。经NSE免疫组织化学显示，12d培养物中神经元约占89％。P19诱导的神经元培养12d，神经元突起经MAP-2染色呈阳性反应。TH免疫组织化学显示，神经元中TH染色阳性神经元约占0.8％。组织化学显示．神经元中AchE染色阳性神经元约占71．09％。结论 P19细胞在体外经定向诱导分化可以产生较高比例的神经元。     

N-甲基-D-天冬氨酸受体在大鼠丘脑前核和扣带后回的表达

目的探讨大鼠丘脑前核和扣带后回内N-甲基-D-天冬氨酸受体NMDAR2A,NMDAR2B及NMDAR1(NR2A.NR2B及NR1)mRNA的表达,从形态学角度为系统研究丘脑前核的功能提供了依据。方法应用原位杂交方法检测丘脑前核及扣带后回内NR2A,NR2B以及NR1 mRNA的表达。结果原位杂交阳性反应产物为棕黄色,主要分布在神经元核周围胞浆中,胞核基本不着色。在丘脑前核,阳性神经元分布较密集,细胞形态较一致。扣带后回皮质阳性神经元在在大脑皮质不同部位分布有差异,分子层染色最弱,外颗粒层密集分布且染色增强,余四层较外颗粒层松散,染色细胞减少。结论 NR2A,NR2B以及NR1广泛分布在丘脑前核和扣带后回。     

NADPH-diaphorase-positive neurons and pathways in the brain of the frog Rana esculenta

We described the NADPH-diaphorase-containing neurons and fibres in the brain of the frog Rana esculenta. In the telencephalon stained cells occurred in the olfactory bulb, all subdivisions of the pallium, the diagonal band, the medial septum and the striatum. The olfactory glomeruli showed the most intense enzyme reaction. The neuropil of the accessory olfactory bulb was also heavily stained and this staining extended to the rostral diencephalon through the ventral lateral pallium. Fibre staining was less intense in the medial pallium and the medial septum. In the diencephalon, NADPH-diaphorase staining was concentrated in the middle third of this part of the brain. The stained cells were embedded in a dense network of thin, stained fibres and terminals in the lateral anterior and central thalamic nuclei. Faintly stained cells were present also in the posterior preoptic nucleus, anterior thalamic nucleus, nucleus of Bellonci, corpus geniculatum thalamicum and the suprachismatic nucleus. In the mesencephalon, heavily stained cells occurred in the nucleus profundus mesencephali, anterodorsal, anteroventral and especially in the posterodorsal tegmental nuclei. Neuronal staining was less intense in the optic tectum and the torus semicircularis. Thick, intensely stained fibres occupied the lateral part of the tegmentum and the 7th layer of the tectum. A loose network of thin fibres occupied the periventricular area and all tegmental nuclei. In the rhombencephalon, the reticular nuclei and the inferior raphe nucleus showed the most intense staining, while some cells in the nucleus of the solitary tract and the dorsal column nuclei were less intensely stained. Heavy staining of fibres was characteristic of the spinal trigeminal tract, the solitary tract and the reticulospinal pathway.     

NADPH-diaphorase-positive neurons and pathways in the brain of the frog Rana esculenta

We described the NADPH-diaphorase-containing neurons and fibres in the brain of the frog Rana esculenta. In the telencephalon stained cells occurred in the olfactory bulb, all subdivisions of the pallium, the diagonal band, the medial septum and the striatum. The olfactory glomeruli showed the most intense enzyme reaction. The neuropil of the accessory olfactory bulb was also heavily stained and this staining extended to the rostral diencephalon through the ventral lateral pallium. Fibre staining was less intense in the medial pallium and the medial septum. In the diencephalon, NADPH-diaphorase staining was concentrated in the middle third of this part of the brain. The stained cells were embedded in a dense network of thin, stained fibres and terminals in the lateral anterior and central thalamic nuclei. Faintly stained cells were present also in the posterior preoptic nucleus, anterior thalamic nucleus, nucleus of Bellonci, corpus geniculatum thalamicum and the suprachismatic nucleus. In the mesencephalon, heavily stained cells occurred in the nucleus profundus mesencephali, anterodorsal, anteroventral and especially in the posterodorsal tegmental nuclei. Neuronal staining was less intense in the optic tectum and the torus semicircularis. Thick, intensely stained fibres occupied the lateral part of the tegmentum and the 7th layer of the tectum. A loose network of thin fibres occupied the periventricular area and all tegmental nuclei. In the rhombencephalon, the reticular nuclei and the inferior raphe nucleus showed the most intense staining, while some cells in the nucleus of the solitary tract and the dorsal column nuclei were less intensely stained. Heavy staining of fibres was characteristic of the spinal trigeminal tract, the solitary tract and the reticulospinal pathway.     

Distribution of NADPH diaphorase-containing neurons in the pigeon central nervous system

The aim of the present study was to determine the distribution of nitric oxide-synthesizing neurons in the pigeon brain and spinal cord. Tissue sections were stained for reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d). In the telencephalon, intensely stained neurons with dendrites extending distally were seen in most regions. The ectostriatum was characterized by intensely and diffusely stained neuropil. In the diencephalon, intensely positive neurons were seen in the lateral hypothalamic region and lateral mammillary nucleus. In the mesencephalon, intensely stained, multipolar neurons were abundantly scattered in the central gray, nucleus intercollicularis, reticular formation, nucleus tegmenti pedunculo-pontinus, pars compacta, area ventralis of Tsai, and ansa lenticularis. In the rhombencephalon, positively-stained neurons were found in the pontine nuclei and reticular formation. The cerebellar cortex, except for Purkinje cells, was a preferential region for NADPH-d activity. Positive end-bulbs made contact on somata in the nucleus magnocellularis cochlearis. In the spinal cord, NADPH-d positive neurons were seen in layer II and the marginal nucleus. Our results demonstrated that the distribution of NADPH-d-containing neurons in the pigeon brain and spinal cord is more complex than in other avian species. Our findings indicate that NADPH-d-containing neurons are present in several sensory pathways, including olfactory, visual, auditory, and somatosensory tracts, although some nuclei in each system did not show NADPH-d activity. The wide distribution of NADPH-d activity in the pigeon CNS suggests that nitric oxide modulates sensory transmission in avian central nervous system.     

IMMUNOHISTOCHEMICAL DISTRIBUTION OF CANNABINOID CB 1 RECEPTOR IN THE RAT CENTRAL NERVOUS SYSTEM

IMMUNOHISTOCHEMICAL DISTRIBUTION OF CANNABINOID CB 1 RECEPTOR IN THE RAT CENTRAL NERVOUS SYSTEM@邹冈     

Histochemical distribution of zinc in the brain of the zebra finch (Taenopygia guttata)

The distribution of zinc was studied in the brain of the zebra finch (Taenopygia guttata) by means of the selenium histochemical method. A specifie pattern was seen, which usually correlated with the main known architectonic subdivisions. In addition, a few as yet unidentified structures were observed. In the telencephalon, the pallial components were stained with moderate to strong intensity. The only exceptions were the hyperstriatum intercalatus superior, a small medial area in the hyperstriatum accessorium and in the dorsolateral cortex, and the dorsomedial part of the hippocampal complex, which were virtually devoid of staining. Staining of the dorsal ventricular ridge components varied considerably. The archistriatum, the nucleus accumbens, the nucleus of the stria terminalis, the hyperstriatum ventrale and the lateral septum showed moderate to strong staining. The medial septum was weakly stained. The neostriatum showed a rather complex pattern of staining with unstained areas, such as the magnocellular nucleus of the anterior neostriatum, and other parts intensely stained, especially in its caudal region. Both paleostriatii primitivum and augmentatum showed a rostro-caudal gradient that was increasingly stained. We also observed an intensely stained area ventral to the fasciculus prosencephali lateralis and lateral to the tractus septomesencephalicus, a weakly to moderately stained band ventral to the lobus parolfactorius, an intensely stained zone along the lateral ventricle in the hyperstriatum ventrale, and an unstained almond-shaped nucleus in the lateral hyperstriatum ventrale. In the diencephalon, the hypothalamus showed a moderate to strong, rather uniform staining, whereas the thalamus was usually weakly to moderately stained, with the exception of a few unstained nuclei. Only the lateral nucleus of the habenula was stained, and with strong intensity. Most of the mesencephalon stained rather uniformly with a moderate to strong intensity. The most intense staining was seen in the substantia grisea centralis, the substantia grisea et fibrosa periventricularis, the torus semicircularis and the nucleus intercollicularis. The tectum opticum was virtually devoid of stain except for two light bands in the stratum griseum et fibrosum superficiale. The formatio reticularis was moderately stained. All the other structures were either weakly stained or unstained. Some staining was seen in the Purkinje and the granular layers of tha cerebellum, as well as around its internal nuclei. The pons and the medulla oblongata showed an overall moderate to intense staining, with the exception of a few unstained nuclei. When compared in three bird species belonging to different genera, zinc distribution shows remarkable similarities, despite species, age and methodological differences. The pattern of zinc staining suggests that this element may play an important role in integrative and autonomic functions.     

Histochemical distribution of zinc in the brain of the zebra finch (Taenopygia guttata)

The distribution of zinc was studied in the brain of the zebra finch (Taenopygia guttata) by means of the selenium histochemical method. A specifie pattern was seen, which usually correlated with the main known architectonic subdivisions. In addition, a few as yet unidentified structures were observed. In the telencephalon, the pallial components were stained with moderate to strong intensity. The only exceptions were the hyperstriatum intercalatus superior, a small medial area in the hyperstriatum accessorium and in the dorsolateral cortex, and the dorsomedial part of the hippocampal complex, which were virtually devoid of staining. Staining of the dorsal ventricular ridge components varied considerably. The archistriatum, the nucleus accumbens, the nucleus of the stria terminalis, the hyperstriatum ventrale and the lateral septum showed moderate to strong staining. The medial septum was weakly stained. The neostriatum showed a rather complex pattern of staining with unstained areas, such as the magnocellular nucleus of the anterior neostriatum, and other parts intensely stained, especially in its caudal region. Both paleostriatii primitivum and augmentatum showed a rostro-caudal gradient that was increasingly stained. We also observed an intensely stained area ventral to the fasciculus prosencephali lateralis and lateral to the tractus septomesencephalicus, a weakly to moderately stained band ventral to the lobus parolfactorius, an intensely stained zone along the lateral ventricle in the hyperstriatum ventrale, and an unstained almond-shaped nucleus in the lateral hyperstriatum ventrale. In the diencephalon, the hypothalamus showed a moderate to strong, rather uniform staining, whereas the thalamus was usually weakly to moderately stained, with the exception of a few unstained nuclei. Only the lateral nucleus of the habenula was stained, and with strong intensity. Most of the mesencephalon stained rather uniformly with a moderate to strong intensity. The most intense staining was seen in the substantia grisea centralis, the substantia grisea et fibrosa periventricularis, the torus semicircularis and the nucleus intercollicularis. The tectum opticum was virtually devoid of stain except for two light bands in the stratum griseum et fibrosum superficiale. The formatio reticularis was moderately stained. All the other structures were either weakly stained or unstained. Some staining was seen in the Purkinje and the granular layers of tha cerebellum, as well as around its internal nuclei. The pons and the medulla oblongata showed an overall moderate to intense staining, with the exception of a few unstained nuclei. When compared in three bird species belonging to different genera, zinc distribution shows remarkable similarities, despite species, age and methodological differences. The pattern of zinc staining suggests that this element may play an important role in integrative and autonomic functions.     

Characterization of neuronal subsets surrounded by perineuronal nets in the rhesus auditory brainstem

The distribution of perineuronal nets and the potassium channel subunit Kv3.1b was studied in the subdivisions of the cochlear nucleus, the medial nucleus of the trapezoid body, the medial and lateral superior olivary nuclei, the lateral lemniscal nucleus and the inferior colliculus of the rhesus monkey. Additional sections were used for receptor autoradiography to visualize the patterns of GABAA and GABAB receptor distribution. The Kv3.1b protein and perineuronal nets [visualized as Wisteria floribunda agglutinin (WFA) binding] were revealed, showing corresponding region-specific patterns of distribution. There was a gradient of labelled perineuronal nets which corresponded to that seen for the intensity of Kv3.1b expression. In the cochlear nucleus intensely and faintly stained perineuronal nets were intermingled, whereas in the medial nucleus of the trapezoid body the pattern changed to intensely stained perineuronal nets in the medial part and weakly labelled nets in its lateral part. In the inferior colliculus, intensely labelled perineuronal nets were arranged in clusters and faintly labelled nets were arranged in sheets. Using receptor autoradiography, GABAB receptor expression in the anterior ventral cochlear nucleus was revealed. The medial part of the medial nucleus of the trapezoid body showed a high number of GABAA binding sites whereas the lateral part exhibited more binding sites for GABAB. In the inferior colliculus, we found moderate GABAB receptor expression. In conclusion, intensely WFA-labelled structures are those known to be functionally involved in high-frequency processing.     

Developmental changes of NADPH-diaphorase positive structures in the isthmic nuclei of the chick

Nicotine adenine dinucleotide phosphate-dia- phorase staining was used to study nitric oxide synthase activity and distribution in the midbrain visual structures of white leghorn chick embryos and post-hatched chickens. Enzyme staining first appeared in the isthmic region at the tenth embryonic day (E10) in the neuropil of the nucleus isthmi, pars parvocellularis. At E11 faint enzyme positivity appeared also in the nucl. isthmi pars magnocellularis, the nucl. semilunaris and the nucl. isthmo-opticus. The staining intensity of the isthmic nuclei dramatically increased between the 12.5th and the 13th days of incubation. The nucl. isthmi, pars parvocellularis showed the strongest enzyme reaction throughout embryonic life. A day before hatching all the isthmic nuclei were heavily stained, however, nicotine adenine dinucleotide phosphate-diaphorase-positive cells occurred exclusively in the nucleus isthmo-opticus. In the tectum opticum, intensely stained cells occupied the stratum fibrosum et griseum superficiale. The layer containing the projection neurons to the isthmo-optic nucleus was unstained. In the isthmic region, the intensity of staining surpassed that of the tectum and reached its maximum at E17 and then slowly decreased till the end of the experimental period (120 days post-hatched). The tractus isthmo-opticus showed nicotine adenine dinucleotide phosphate-diaphorase activity throughout the investigated period of life of the chicken, but the tractus tectoisthmo-opticus was unstained. Our results suggest that in the isthmic nuclei, nicotine adenine dinucleotide phosphate-diaphorase-positive neurons occur only in the isthmo-optic nucleus and optic tectum. The other positively stained structures are the fibers and terminals of tectal cells. In most brain areasnicotine adenine dinucleotide phosphate-diaphorase indicates nitric oxide synthase that produces nitric oxide. The transient appearance of this molecule is probably necessary for neuronal differentiation or the establishment of synaptic connections in the isthmic nuclei, and these developmental changes are under the control of the optic tectum.     

Developmental changes of NADPH-diaphorase positive structures in the isthmic nuclei of the chick

Nicotine adenine dinucleotide phosphate-diaphorase staining was used to study nitric oxide synthase activity and distribution in the midbrain visual structures of white leghorn chick embryos and post-hatched chickens. Enzyme staining first appeared in the isthmic region at the tenth embryonic day (E10) in the neuropil of the nucleus isthmi, pars parvocellularis. At E11 faint enzyme positivity appeared also in the nucl. isthmi pars magnocellularis, the nucl. semilunaris and the nucl. isthmo-opticus. The staining intensity of the isthmic nuclei dramatically increased between the 12.5th and the 13th days of incubation. The nucl. isthmi, pars parvocellularis showed the strongest enzyme reaction throughout embryonic life. A day before hatching all the isthmic nuclei were heavily stained, however, nicotine adenine dinucleotide phosphate-diaphorase-positive cells occurred exclusively in the nucleus isthmo-opticus. In the tectum opticum, intensely stained cells occupied the stratum fibrosum et griseum superficiale. The layer containing the projection neurons to the isthmo-optic nucleus was unstained. In the isthmic region, the intensity of staining surpassed that of the tectum and reached its maximum at E17 and then slowly decreased till the end of the experimental period (120 days post-hatched). The tractus isthmo-opticus showed nicotine adenine dinucleotide phosphate-diaphorase activity throughout the investigated period of life of the chicken, but the tractus tectoisthmo-opticus was unstained. Our results suggest that in the isthmic nuclei, nicotine adenine dinucleotide phosphate-diaphorase-positive neurons occur only in the isthmo-optic nucleus and optic tectum. The other positively stained structures are the fibers and terminals of tectal cells. In most brain areas nicotine adenine dinucleotide phosphate-diaphorase indicates nitric oxide synthase that produces nitric oxide. The transient appearance of this molecule is probably necessary for neuronal differentiation or the establishment of synaptic connections in the isthmic nuclei, and these developmental changes are under the control of the optic tectum.     

LIGHT AND ELECTRON MICROSCOPIC OBSERVATIONS OF HEPATIC HEMATOPOIESIS OF HUMAN FETUSES

Liver tissues obtained from 78 human fetuses between 5 weeks and 10 months of gestation were observed with light and electron microscope. In the third month of gestation hepatic megakaryocytopoiesis reached the maximum level and then appeared to decline. At the maximum level immature to mature megakaryocytes were usually smaller than those from adult bone marrow. They were observed between growing liver cells, within hepatic sinusoids and in the mesenchyme surrounding the liver parenchyma in the given order of frequency. Immature megakaryocytes were characterized by the presence of both demarcation vesicles or tubules and a few small round specific granules. The nucleus was mononuclear but frequently indented. With cell maturation, the megakaryocytes were stained intensely by PAS-technique. The nuclei were multilobulated. Large cytoplasm was dominated by numerous specific granules and well-developed demarcation membrane system. Some megakaryocytes passed through the sinusoidal wall and possibly to hepatic sinusoids. These observations indicate that the main site of hepatic megakaryocytopoiesis may be extravascular and more likely between liver cells. Small megakaryocytes in fetal livers are discussed in relation to the occurrence of similar megakaryocytes in various blood dyscrasias.