Abstract：
The cell intrinsic mechanisms directing peripheral nerve regeneration have remained largely understudied, thus limiting our understanding of these processes and constraining the advancement of novel clinical therapeutics. The use of primary adult rat dorsal root ganglion (DRG) neurons cultured in vitro is well established. Despite this, these cells can be challenging to culture and have so far not been amenable to robust transfection or live-cell imaging. The ability to transfect these cells with fluorescent plasmid constructs to label subcellular structures, combined with high resolution time-lapse imaging has the potential to provide invaluable insight into how peripheral neurons coordinate their regenerative response, and which specific cellular structures are involved in this process. Here we describe a protocol that facilitates transfection and subsequent live-imaging of adult rat DRG neurons.
摘要：
指导周围神经再生的细胞内在机制在很大程度上仍未被研究。从而限制了我们对这些过程的理解，并限制了新型临床疗法的发展。体外培养的原代成年大鼠背根神经节（DRG）神经元的使用已经确立。尽管如此,这些细胞可能对培养具有挑战性，并且到目前为止还不适合进行强大的转染或活细胞成像。用荧光质粒构建体转染这些细胞以标记亚细胞结构的能力，结合高分辨率延时成像有可能提供宝贵的洞察力，以了解周围神经元如何协调其再生反应，以及该过程涉及哪些特定的细胞结构。在这里，我们描述了一种促进成年大鼠DRG神经元的转染和随后的活体成像的方案。
