Ruthazer Edward S, Schohl Anne, Schwartz Neil, Tavakoli Aydin, Tremblay Marc, Cline Hollis T
Cold Spring Harb Protoc. 2013 Aug 1;2013(8):771-5. doi: 10.1101/pdb.prot076471.
Individual neurons in the developing nervous system of Xenopus laevis can be visualized by the targeted delivery of a fluorophore. The fluorophore can be delivered as a fluorescent dye or DNA that encodes a fluorescent protein. Local iontophoresis is a method that works well for transfer of fluorescent dye to retinal ganglion cells (RGCs) in the eye, but it does not give a high yield for delivery of DNA. This is largely because the degree of pigmentation of the eyes, even in albino strains, makes it difficult to visualize RGC somata during pipette positioning. Bulk retinal electroporation is a better approach for delivery of plasmid DNA to RGC. The method described here works best in tadpoles older than stage 42.
通过荧光团的靶向递送，可以观察到非洲爪蟾发育中的神经系统中的单个神经元。荧光团可以作为荧光染料或编码荧光蛋白的DNA进行递送。局部离子电渗法是一种将荧光染料转移到眼中视网膜神经节细胞（RGC）的有效方法，但它在递送DNA时产量不高。这主要是因为即使是白化病品系，眼睛的色素沉着程度也使得在移液管定位过程中难以观察到RGC胞体。批量视网膜电穿孔是将质粒DNA递送至RGC的更好方法。此处描述的方法在42期以上的蝌蚪中效果最佳。
