Isolation of Transcriptomic-Quality Total RNA from Mouse Spinal Cords.
Isolation of Transcriptomic-Quality Total RNA from Mouse Spinal Cords.
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DOI：
10.1002/cpz1.338
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发表时间：
2022-01
期刊：
影响因子：
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通讯作者：
Kaplan BLF
中科院分区：
文献类型：
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作者：
Stokes JV;Nicaise AJ;Frodella CM;Varela-Stokes AS;Thompson T;Kaplan BLF
评估脊髓中的细胞，蛋白质和总RNA对于促进我们对神经炎症和神经退行性疾病的理解至关重要。例如，在实验性自身免疫性脑脊髓炎（EAE）模型中，免疫细胞浸润脊髓，通常用于研究多发性硬化症。因此，评估总RNA以确定脊髓中的神经元和炎症特征是有价值的。此外，RNA谱可用于解读药物或化学品对神经炎症和神经退行性疾病如EAE的影响。本方案和视频的目的是描述和证明脊髓从小鼠脊柱中排出，并使用液氮对脊髓进行均质化，以实现最佳RNA分离。虽然我们提出了这种方法与脊髓从EAE小鼠，该技术是广泛适用的，包括RNA分离从脊髓的健康小鼠。这些步骤的正确执行对于在使用市售试剂盒进行最终分离时获得足够的转录组学质量的脊髓RNA产量至关重要。从脊柱中分离脊髓分离用钝头针的制备使用液氮的脊髓匀浆RNA纯度、定量和完整性的评估
Assessing cells, proteins, and total RNA in the spinal cord is vital for advancing our understanding of neuroinflammation and neurodegenerative diseases. For instance, immune cells infiltrate the spinal cord in the experimental autoimmune encephalomyelitis (EAE) model, commonly used to study multiple sclerosis. Thus, it is valuable to assess total RNA to determine the neuronal and inflammatory profiles in the spinal cord. Further, RNA profiles are useful for deciphering the effects of drugs or chemicals on neuroinflammation and on neurodegenerative diseases such as EAE. The purpose of this protocol and video is to describe and demonstrate the expulsion of the spinal cord from the mouse spinal column and homogenization of the spinal cord using liquid nitrogen for optimal RNA isolation. Although we present this method with spinal cords from EAE mice, the technique is broadly applicable, including RNA isolation from the spinal cords of healthy mice. Proper performance of these steps is critical to achieving a sufficient yield of transcriptomic-quality spinal cord RNA when combined with final isolation using commercially available kits. Isolation of spinal cords from the spinal column Preparation of blunt-end needle for spinal cord isolation Spinal cord homogenization using liquid nitrogen Assessment of RNA purity, quantification, and integrity