生物双重显微结构三维重建图象显示及畸变补偿方法

本文介绍计算机三维重建技术的两个进展:双重显微结构三维重建图象的显示技术及三维重建图象显示畸变的补偿方法。 1.生物双重显微结构的三维显示方法 有些生物的神经核团具有双重结构,例如蛤蚧的中脑峡核,它由大细胞部(Imc)和小细胞部(Ipc)两部分组成,仅Imc与视觉有关,Ipc似乎既无视觉功能也无听觉功能。但Imc与Ipc紧挨着,且有部分结构交迭在     

生物显微结构连续切片图象的象素地址矢量表达和三维重建信息加工

本文主要介绍一种适用于生物显微结构连续切片三维重建的图象表达方法——象素地址矢量法及其相应的图象信息加工方法:三维重建,三维模型绕任选空间轴旋转和用软件重新剖切.与常用的图象矩阵表达法相比,可以较多地节省序列图象所占的数据容量,便于在微机系统上对多种形态的显微结构进行三维重建信息加工.当对较大的图象进行分解与整合处理时,还避免了采用图象矩阵表达法可能遇到的边界问题.由于数据容量的减少,提高了图象处理的速度.这种图象表达及信息加工方法对包括生物活体时间序列图象在内的多种序列图象加工可能都是适用的.     

GABA能神经元在鸽峡核中的分布

本文用免疫细胞化学技术研究了r-氨基丁酸(GABA)阳性神经元在家鸽(Columba livia)峡核中的分布特点.实验结果表明,GABA阳性神经元在峡核大细胞部(Imc)呈均匀分布.约占细胞总数的41%,而在峡核小细胞部(Ipc)GABA阳性神经元仅位于核嘴内侧部.     

家鸽血管活性肠肽阳性神经元在顶盖和峡核中的分布及形态特征

用免疫细胞化学技术研究了血管活性肠肽(VIP)阳性神经元在家鸽(Columba livia)视顶盖和峡核中的分布,实验结果表明.在顶盖中VIP阳性神经元主要分布在Ⅱ_1层.并有少数VIP阳性神经元位于Ⅱ_1和Ⅲ层,在峡核大细胞部(Ime)和峡核小细胞部(Ipc),VIP阳性神经元基本呈均匀分布,在Imc和Ipc内分别占细胞总数的80%和50%.     

计算机三维重建华虻复眼性特异小网膜细胞结构

本文在计算机三维重建的基础上讨论了华虻复眼小网膜性特异结构与功能的关系,三维模型展示了中央小网膜细胞和部分外周小网膜细胞的空间构型及相互关系.在生物组织连续切片三维重建的对位技术方面,摸索了一套方法.保证了在Cromemco微机系统上实现对华虻复眼小网膜细胞的三维重建.证实了复眼光感受器性特化区R_7和R_8为并行排列的形式,并与非性特化区的R_7、R_8进行了比较.     

人外周血单个核细胞与脐带间充质干细胞共培养的AFM研究

采用原子力显微镜与倒置显微镜在细胞层次上观察了人外周单个核细胞(PBMCs)与同种异源脐带间充质干细胞(hUC-MSCs)共培养的过程,并在单细胞水平上分析了共培养前后人外周单个核细胞的形貌和生物物理性质。结果发现:共培养后贴壁人外周单个核细胞的形态发生了很大的改变,并且表面分布着大小不一的颗粒状聚合物。利用AFM高空间分辨的力位移曲线测量系统,发现共培养72h后培养上清中人外周单个核细胞、贴壁的人外周单个核细胞的粘滞力分别是单纯培养72h的人外周单个核细胞的2倍、5倍,而细胞的硬度分别是单纯培养人外周单个核细胞的1.5倍、2倍。CCK-8检测提示,共培养过程中,干细胞的生长与外周血单个核细胞的生长出现了竞争作用。通过AFM探测人外周单个核细胞与脐带间充质干细胞共培养的可视化数据,有助于更好地了解间充质干细胞与外周血单个核细胞的相互作用。     

蛤蚧豆状核的结构及其与顶盖前端的纤维联系

运用Nissl法和辣根过氧化物酶(horseradish peroxidase,HRP)追踪标记技术,研究蛤蚧(Gekko gecko)豆状核的结构及其与顶盖前端的纤维联系。Nissl染色显示,蛤蚧豆状核细胞大小没有明显差别,由背内侧细胞密集部和腹外侧细胞稀疏部组成。将HRP注射于顶盖前端,结果豆状核背内侧部和腹外侧部分别接受同侧顶盖前端脑室内、外侧纤维的传入,核内标记有浓密的神经丛和大量纤维末梢,并在该核腹外侧部及其邻近区域发现少量大胞体标记细胞。推测豆状核腹外侧部的大胞体细胞及其邻近区域的大胞体细胞可能具有相同的功能,且该核可能形成离顶盖通路和副视系统相联系的交通要道。     

蛤蚧视觉神经核团的细胞形态结构及功能的研究

本研究发现,蛤蚧视觉神经核团有视顶盖（OT）、峡核（NI）、基底视束核（nBOR）、豆状核（LM）、中脑深部核（NPM）、圆核（NR）、前背侧室嵴（ADVR）和皮质加厚区（Pth）等,其中NI和ADVR两核团的体积最大。视觉核团中有各种形状的细胞形态,其中梨形和梭形细胞占的比例较大。神经核团的细胞直径为6～30μm,其中以15～28μm最多。在ADVR和Pth核团中有细胞丛簇存在,其它核团尚未发现有这样的结构。各神经核团问和核团内有广泛而复杂的纤维联系。蛤蚧有关视觉神经核团除具有视觉功能外,可能还与听觉、触觉、嗅觉和平衡感觉等功能有关。     

大鼠p75NTR荧光真核表达载体的构建及其在HEK293细胞中的表达

目的:构建大鼠p75神经营养素受体(p75 neurotrophin receptor,p75NTR)cDNA序列的绿色荧光真核表达载体并鉴定其在人胚肾293(human embryo kidney 293,HEK293)细胞中的表达.方法:采用PCR方法从含野生型大鼠p75NTR的pDC316-RP75质粒中扩增目的片段,经EcoRⅠ和SaⅡ双酶切,定向克隆于pEGFP-N1质粒中,构建绿色荧光真核表达栽体pEGFP-N1-RP75,经酶切及测序鉴定后,通过脂质体转染HEK293细胞,激光共聚焦及免疫组织化学法鉴定大鼠p75NTR的表达.结果:重组质粒经酶切鉴定和序列分析证实含有大鼠p75NTR的编码序列,转染后经激光共聚焦显微镜及免疫组织化学染色观察表明重组质粒能够在HEK293细胞中表达出具有活性的大鼠p75NTR片段.结论:大鼠p75NTR绿色荧光真核表达栽体构建成功并可在HEK293细胞中表达,为进一步研究奠定了基础.     

东亚飞蝗脑的形态学观察及三维重建

本文主要研究东亚飞蝗Locusta migratoria manilansis(Meyen)脑部的形态结构及其三维重建模型。采用石蜡包埋切片,在光镜下观察了东亚飞蝗脑部的形态结构,其由前脑、中脑和后脑3部分组成。为了获得整只蝗虫的连续、完整的图像数据集,采用冰冻切片技术将冰冻包埋剂(OCT)包埋的飞蝗成虫做连续切片。然后利用图像处理方法对飞蝗脑部的连续切片进行配准、分割,再用三维重建软件Image-Pro Plus(IPP)对分割后的脑部二维图像序列进行三维重建,构建出的飞蝗脑部三维结构模型可以任意旋转,能从不同角度观察。其结果为蝗虫生理和防蝗治蝗提供科学依据。     

Response properties of visual neurons in the turtle nucleus isthmi

The optic tectum holds a central position in the tectofugal pathway of non-mammalian species and is reciprocally connected with the nucleus isthmi. Here, we recorded from individual nucleus isthmi pars parvocellularis (Ipc) neurons in the turtle eye-attached whole-brain preparation in response to a range of computer-generated visual stimuli. Ipc neurons responded to a variety of moving or flashing stimuli as long as those stimuli were small. When mapped with a moving spot, the excitatory receptive field was of circular Gaussian shape with an average half-width of less than 3°. We found no evidence for directional sensitivity. For moving spots of varying sizes, the measured Ipc response-size profile was reproduced by the linear Difference-of-Gaussian model, which is consistent with the superposition of a narrow excitatory center and an inhibitory surround. Intracellular Ipc recordings revealed a strong inhibitory connection from the nucleus isthmi pars magnocellularis (Imc), which has the anatomical feature to provide a broad inhibitory projection. The recorded Ipc response properties, together with the modulatory role of the Ipc in tectal visual processing, suggest that the columns of Ipc axon terminals in turtle optic tectum bias tectal visual responses to small dark changing features in visual scenes.     

A Golgi study of the isthmic nuclei in the pigeon (Columba Iivia)

Summary The isthmic nuclei of the pigeon were studied with the use of three different Golgi techniques. The nucleus isthmo-opticus (IO) consists of a single cell type in which all dendrites of one neuron take the same direction and ramify at identical distances from the perikaryon to form dense dendritic arborizations. The cell bodies of the IO neurons form two parallel layers. The dendrites of these neurons always extend to the area between the two layers so that the dendritic arborizations of opposite neurons overlap. A model of the cellular organization of the IO was constructed based upon these morphological characteristics. The neurons of the n. isthmi/pars parvocellularis (Ipc) have oval perikarya and long, smooth, infrequently branching dendrites. All neurons except those at the borders of the nucleus show the same dorsoventral orientation in their dendritic arborizations and together with their afferents seem to have a columnar organization. The dendrites of the neurons located at the margin of the nucleus ramify within the Ipc along its border. The n. semilunaris (Slu) consists of neurons with round somata that have on an average three dendrites with small spines. The axons leave the nucleus from the medial side and join the lemniscus lateralis. The neurons of the n. isthmi/pars magnocellularis (Imc) comprise a generalized isodendritic type resembling the cells of the reticular formation. Axons from the tectum penetrate the nucleus, making numerous en-passant contacts with several neurons.     

用羰花青荧光染料追踪家鸽顶盖与中脑核团的神经连接

本文用亲脂质荧光染羰花青(Carbocyanine)的衍生物——1.1’一二(十八烷基)—3,3,3,’3’—四甲基吲哚羰花青高氯酸盐(1,1’,dioctadecyl—3,3,3’,3’—tetramethyli-ndocarbocyanine perchlorate,dil),追踪了家鸽(Columba livia)顶盖深层与中脑核团的神经连接.结果表明,顶盖深层接收峡核大细胞部(Nucleus isthmi pars magnocellularis,Imc)以及动眼神核腹部(Nucleus nervi oculomotorii,pars ventralis,OMv)中脑外侧核背部(Nucleus mesencephalicus lateralis,pars dorsalis,MLd)的神经投射,本文还对这些结果与以前用类霍乱原-HRP(CB HRP)方法所得结果进行了比较分析.     

Spatially Reciprocal Inhibition of Inhibition within a Stimulus Selection Network in the Avian Midbrain

Reciprocal inhibition between inhibitory projection neurons has been proposed as the most efficient circuit motif to achieve the flexible selection of one stimulus among competing alternatives. However, whether such a motif exists in networks that mediate selection is unclear. Here, we study the connectivity within the nucleus isthmi pars magnocellularis (Imc), a GABAergic nucleus that mediates competitive selection in the midbrain stimulus selection network. Using laser photostimulation of caged glutamate, we find that feedback inhibitory connectivity is global within the Imc. Unlike typical lateral inhibition in other circuits, intra-Imc inhibition remains functionally powerful over long distances. Anatomically, we observed long-range axonal projections and retrograde somatic labeling from focal injections of bi-directional tracers in the Imc, consistent with spatial reciprocity of intra-Imc inhibition. Together, the data indicate that spatially reciprocal inhibition of inhibition occurs throughout the Imc. Thus, the midbrain selection circuit possesses the most efficient circuit motif possible for fast, reliable, and flexible selection.     

Generating oscillatory bursts from a network of regular spiking neurons without inhibition

Avian nucleus isthmi pars parvocellularis (Ipc) neurons are reciprocally connected with the layer 10 (L10) neurons in the optic tectum and respond with oscillatory bursts to visual stimulation. Our in vitro experiments show that both neuron types respond with regular spiking to somatic current injection and that the feedforward and feedback synaptic connections are excitatory, but of different strength and time course. To elucidate mechanisms of oscillatory bursting in this network of regularly spiking neurons, we investigated an experimentally constrained model of coupled leaky integrate-and-fire neurons with spike-rate adaptation. The model reproduces the observed Ipc oscillatory bursting in response to simulated visual stimulation. A scan through the model parameter volume reveals that Ipc oscillatory burst generation can be caused by strong and brief feedforward synaptic conductance changes. The mechanism is sensitive to the parameter values of spike-rate adaptation. In conclusion, we show that a network of regular-spiking neurons with feedforward excitation and spike-rate adaptation can generate oscillatory bursting in response to a constant input.     

TOPOGRAPHICAL AND SUBCELLULAR DISTRIBUTION OF CHOLINE ACETYLTRANSFERASE AND GLUTAMATE DECARBOXYLASE IN PIGEON OPTIC TECTUM

Abstract— The distribution of choline acetyltransferase (ChAT) and glutamate decarboxylase (GAD) in different layers of the pigeon optic tectum and in some nuclei of the optic lobe have been investigated. About 40% of GAD and 25% of ChAT were found in the superficial part of tectum, but negligible activity was found in the stratum opticum. The highest GAD activity was found in layers 3-7 (according to the nomenclature of C ajal , 1911) with a peak in layer 4. ChAT activity peaked in layers 3, 5. 8 and 10/11. Its distribution correlated well with the staining pattern of AChE, particularly in the superficial part of the tectum. The distribution of ChAT and GAD did not change significantly 4 weeks after enucleation. ChAT and GAD activities were high in the nucleus isthmi, pars parvocellularis (Ipc). The activity of GAD was also high in the nucleus intercollicularis (ICo), the other nuclei showed less activity of both enzymes.     

两栖类峡核的顶盖外投射

用辣根过氧化物酶法研究了中华大蟾蜍峡核的顶盖外投射。结果指出：（１）中脑脚盖前背核、中脑深核和表层峡网核投向双侧峡核,对侧投射经过Ｖｅｌｉ交叉;（２）这些脚盖核投向整个峡核,其间没有区域对应关系,讨论了这些投射的可能功能意义。     

家鸽峡核神经元的形态

本文采用Golgi-Cox浸染法,研究了家鸽峡核神经元的形态。按其主树突的分布图式,可将鸽峡核神经元分成三类:(1)单极神经元,(2)双极神经元和(3)多极神经元。峡核两部分(Imc和Ipc)的神经元在胞体大小、树突粗细及其伸展趋势上均有明显差别。