FLUORESCENT LATEX MICROSPHERES AS A RETROGRADE NEURONAL MARKER FOR INVIVO AND INVITRO STUDIES OF VISUAL-CORTEX
FLUORESCENT LATEX MICROSPHERES AS A RETROGRADE NEURONAL MARKER FOR INVIVO AND INVITRO STUDIES OF VISUAL-CORTEX
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DOI：
10.1038/310498a0
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发表时间：
1984-01-01
期刊：
影响因子：
64.8
通讯作者：
DREYER, WJ
中科院分区：
文献类型：
--
作者：
KATZ, LC;BURKHALTER, A;DREYER, WJ
利用各种物质（如酶、凝集素、合成荧光化合物）的轴突逆行运输已经获得了许多关于神经元通路组织的信息。每种类型的逆行跟踪器都有自己的一组属性，这些属性定义了它可以处理的问题的范围1 - 3。我们在这里描述了一种新的逆行示踪剂，罗丹明标记的荧光乳胶微球（直径0.02-0.2µm），与其他可用的体内和体外示踪剂相比，它具有明显的优势。当注射到脑组织中时，这些微球几乎没有扩散，因此产生小而清晰的注射部位。一旦转运回神经元体，标签在体内至少持续10周，固定后至少持续1年。通过脑皮层切片制备中逆行标记细胞的细胞内记录和染色来评估微球没有明显的细胞毒性或光毒性。该方法进一步用于可视化和比较猫视觉皮质切片中不同投影模式的神经元，发现内在轴突和树突的模式存在显著差异。微球的这些特性为在切片和分离细胞培养中鉴定的投射神经元的解剖和生理研究开辟了新的途径。
The use of retrograde axonal transport of various substances (for example, enzymes, lectins, synthetic fluorescent compounds) has yielded much information on the organization of neuronal path-ways. Each type of retrograde tracer has its own set of attributes which define the scope of problems it can address1–3. We describe here a new class of retrograde tracer, rhodamine-labelled fluorescent latex microspheres (0.02–0.2 µm diameter), which have distinct advantages over other available tracers forin vivoandin vitroapplications. When injected into brain tissue, these microspheres show little diffusion and consequently produce small, sharply defined injection sites. Once transported back to neuronal somata, the label persists for at least 10 weeksin vivoand 1 yr after fixation. Microspheres have no obvious cytotoxicity or phototoxicity as assessed by intracellular recording and staining of retrogradely labelled cells in a cortical brain slice preparation. This approach was further used to visualize and compare, in cat visual cortex slices, neurones with different projection patterns, and revealed significant differences in patterns of intrinsic axons and dendrites. These properties of microspheres open new avenues for anatomical and physiological studies of identified projection neurones in slices as well as in dissociated cell cultures.