van den Pol A N, Gallyas F
Section of Neurosurgery, Yale University Medical School, New Haven, Connecticut 06510.
J Comp Neurol. 1990 Jun 22;296(4):654-73. doi: 10.1002/cne.902960410.
A new esterification-silver approach to the directed staining of the dendritic trees of traumatized neurons is described. Stained neurons compare favorably to those labeled with silver chromate Golgi impregnations in the visualization of dendritic arbors. Cells in all parts of the brain, including the hypothalamus, hippocampus, cerebral cortex, cerebellum, striatum, spinal cord, thalamus, and olfactory bulb, can be detected after focal trauma to that region. Selection of neurons to be stained is made by increasing their affinity for silver with many different types of directed neuronal trauma, including micropipette wounds, pressure, surgical incision, chemical cytotoxicity, and impact trauma. Trauma to one area of the brain results in dendritic arbors staining only in the injured area; other areas of the brain are free of dendritic staining. Injury can be produced either in vivo or in vitro. In vitro injury to neurons allows a high degree of localization and facilitates the analysis of neuronal response to trauma in the absence of complicating factors such as blood flow and secondary injury. The selective affinity for silver staining in this approach is increased very rapidly, allowing detection of traumatized cells fixed minutes after injury. Brains of all mammals used appear to stain similarly, including the rat, mouse, pig, and human. Axons, although labeled less frequently than dendritic arbors, are induced to stain just as rapidly as dendrites. The ability to visualize a large part of the dendritic tree after trauma allows the segregation of neuronal subtypes on the basis of their differential response to injury. Subpopulations of cells in the same area of the brain appear to respond differently to trauma. Differential response of neurons to trauma can easily be detected. For instance, slight variations in trauma can be used to label selectively the major subpopulations of neurons in the hippocampus, including pyramidal cells, interneurons near the pyramidal cell layer, or granule cells. Similarly, neurons of the hypothalamic paraventricular and arcuate nuclei respond to compression trauma much more dramatically than other cell types in the same region of the hypothalamus. Hypothalamic neurons were studied extensively, particularly in regions that are difficult to routinely stain with Golgi impregnations, including the arcuate, paraventricular, supraoptic, and suprachiasmatic nuclei. Trauma to these areas was made in vitro after removal of the brain from the skull, allowing easy access to the ventral surface of the brain.(ABSTRACT TRUNCATED AT 400 WORDS)
本文描述了一种新的酯化-银染色方法,用于对受损伤神经元的树突进行定向染色。在树突分支的可视化方面,染色后的神经元与用铬酸银高尔基浸染法标记的神经元相比毫不逊色。在对大脑的特定区域造成局部创伤后,可以检测到大脑各个部位的细胞,包括下丘脑、海马体、大脑皮层、小脑、纹状体、脊髓、丘脑和嗅球。通过多种不同类型的定向神经元创伤,包括微量移液器损伤、压力、手术切口、化学细胞毒性和撞击创伤,增加神经元对银的亲和力,从而选择要染色的神经元。对大脑的一个区域造成创伤只会导致受伤区域的树突分支染色;大脑的其他区域则没有树突染色。损伤可以在体内或体外产生。对神经元进行体外损伤可以实现高度定位,并有助于在没有诸如血流和继发性损伤等复杂因素的情况下分析神经元对创伤的反应。这种方法中对银染色的选择性亲和力增加得非常迅速,能够检测到损伤后几分钟内固定的受损伤细胞。所有使用的哺乳动物大脑,包括大鼠、小鼠、猪和人类,似乎都有相似的染色效果。轴突虽然比树突分支标记得少,但也能像树突一样迅速被诱导染色。创伤后能够可视化大部分树突树,这使得可以根据神经元对损伤的不同反应来区分神经元亚型。大脑同一区域的细胞亚群对创伤的反应似乎不同。很容易检测到神经元对创伤的不同反应。例如,轻微的创伤差异可用于选择性标记海马体中的主要神经元亚群,包括锥体细胞、锥体细胞层附近的中间神经元或颗粒细胞。同样,下丘脑室旁核和弓状核的神经元对压迫创伤的反应比下丘脑同一区域的其他细胞类型更为显著。对下丘脑神经元进行了广泛研究,特别是在难以用高尔基浸染法进行常规染色的区域,包括弓状核、室旁核、视上核和视交叉上核。在将大脑从颅骨中取出后,对这些区域进行体外创伤,这样便于接触到大脑的腹侧表面。(摘要截断于400字)