Richner Mette, Jager Sara B, Siupka Piotr, Vaegter Christian B
Department of Biomedicine, Aarhus University;
Department of Biomedicine, Aarhus University.
J Vis Exp. 2017 Jan 22(119):55226. doi: 10.3791/55226.
Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may result in damage to the spinal cord caused by the dissection process. Here, we show how the spinal cord can be extruded using hydraulic pressure. Handling time is significantly reduced to only a few minutes, likely decreasing protein damage. The low risk of damage to the spinal cord tissue improves subsequent immunohistochemical analysis. By performing hydraulic spinal cord extrusion instead of traditional laminectomy, the rodents can further be used for DRG isolation, thereby lowering the number of animals and allowing analysis across tissues from the same rodent. We demonstrate a consistent method to identify and isolate the DRGs according to their localization relative to the costae. It is, however, important to adjust this method to the particular animal used, as the number of spinal cord segments, both thoracic and lumbar, may vary according to animal type and strain. In addition, we illustrate further processing examples of the isolated tissues.
传统上,脊髓是通过椎板切除术分离的,即一次打开一个脊椎骨。这既耗时,又可能因解剖过程对脊髓造成损伤。在此,我们展示了如何利用液压挤出脊髓。操作时间显著减少至仅几分钟,可能减少蛋白质损伤。脊髓组织受损风险低,可改善后续免疫组织化学分析。通过进行液压脊髓挤出而非传统椎板切除术,啮齿动物可进一步用于背根神经节(DRG)分离,从而减少动物数量,并允许对同一啮齿动物的不同组织进行分析。我们展示了一种根据DRG相对于肋骨的定位来识别和分离DRG的一致方法。然而,根据所用特定动物调整此方法很重要,因为胸椎和腰椎的脊髓节段数量可能因动物类型和品系而异。此外,我们还举例说明了分离组织的进一步处理方法。