Kasemeier-Kulesa Jennifer C, Lefcort Frances, Kulesa Paul M
Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
CSH Protoc. 2007 Jul 1;2007:pdb.prot4791. doi: 10.1101/pdb.prot4791.
INTRODUCTIONThe peripheral nervous system (PNS) regulates key events within the body including breathing, heart rate, and pain and temperature sensation. In the trunk of the developing embryo, neural crest (NC) cells that follow a ventro-medial pathway form the dorsal root ganglia (DRG) and sympathetic ganglia (SG) of the PNS. The cellular and molecular mechanisms that mediate the formation of the PNS are not completely understood due to the lack of a model system to monitor NC cell migratory behaviors deep within the embryo. Here, we describe a unique sagittal explant culture technique that allows for visualization of DRG and SG formation in the relatively intact chick embryo to assess potential molecules and signals influencing the formation of the PNS. By making a midline cut down the chick antero-posterior axis, each half of the embryo is an explant that can be imaged either from the medial perspective, to reveal events occurring deep within the embryo (SG formation), or from the lateral surface, to image events occurring in the dorso-lateral plane (DRG formation).
引言
周围神经系统（PNS）调节体内的关键活动，包括呼吸、心率以及疼痛和温度感知。在发育中的胚胎躯干中，沿腹内侧路径迁移的神经嵴（NC）细胞形成了PNS的背根神经节（DRG）和交感神经节（SG）。由于缺乏监测胚胎内部深处NC细胞迁移行为的模型系统，介导PNS形成的细胞和分子机制尚未完全明确。在此，我们描述了一种独特的矢状外植体培养技术，该技术可在相对完整的鸡胚中观察DRG和SG的形成，以评估影响PNS形成的潜在分子和信号。通过沿鸡胚前后轴作中线切口，胚胎的每一半均成为一个外植体，既可以从内侧视角成像，以揭示胚胎内部深处发生的事件（SG形成），也可以从侧面成像，以观察背外侧平面发生的事件（DRG形成）。
