Anderson Kevin J, Fugaccia Isabella, Scheff Stephen W
Sanders-Brown Center on Aging, and Department of Anatomy and Neurobiology, University of Kentucky, Lexington, Kentucky 40536-0230, USA.
J Neurotrauma. 2003 Nov;20(11):1223-31. doi: 10.1089/089771503770802899.
In an attempt to label dying neurons in the injured spinal cord, we used the novel fluorescein derivative Fluoro-Jade B, which has been reported to specifically label dead or dying neurons in the brain. Rats and mice were subjected to a moderate level of spinal cord injury using an IH impact device and sacrificed at 1, 2, 4, 7, 14, and 21 days post injury. Spinal cord tissue was processed for Fluoro-Jade B histochemistry and included sections throughout the injured region of the cord. No Fluoro-Jade positive neurons were observed in sections from any time point postinjury at any level of the spinal cord. Instead, Fluoro-Jade labeled astrocytes in uninjured control animals and injured animals. The specificity of astrocytic staining was confirmed by co-localizaton of Fluoro-Jade with glial fibrillary acidic protein. We also subjected a group of rats to a sequential cortical contusion injury and spinal cord injury. Sections from these animals showed numerous Fluoro-Jade positive neurons in the hippocampal formation and thalamus underlying the cortical contusion; however, the staining pattern in the spinal cord was identical to those animals that had received spinal cord injury alone.
为了标记受损脊髓中濒死的神经元,我们使用了新型荧光素衍生物Fluoro-Jade B,据报道它能特异性标记大脑中死亡或濒死的神经元。使用IH撞击装置对大鼠和小鼠造成中度脊髓损伤,并在损伤后1、2、4、7、14和21天处死。对脊髓组织进行Fluoro-Jade B组织化学处理,包括整个脊髓损伤区域的切片。在脊髓任何水平损伤后任何时间点的切片中均未观察到Fluoro-Jade阳性神经元。相反,Fluoro-Jade标记了未受伤对照动物和受伤动物中的星形胶质细胞。通过Fluoro-Jade与胶质纤维酸性蛋白的共定位证实了星形胶质细胞染色的特异性。我们还对一组大鼠进行了连续的皮质挫伤损伤和脊髓损伤。这些动物的切片显示,皮质挫伤下方的海马结构和丘脑中存在大量Fluoro-Jade阳性神经元;然而,脊髓中的染色模式与仅接受脊髓损伤的动物相同。