What fluorescent protein is best for bright, well-defined axonal labeling in culture?
tdTomato is OK, but it’s cytoplasmic and I notice its much brighter in the soma.
My goal is to look at axon degeneration in live cultures (no immunostaining).
ChR2-EYFP from the Ai32 mice looks really nice because it’s membrane-localized. But I don’t want to use ChR2 as the fluor.
What is a good, membrane localized fluor is readily available and looks bright in DRG neurons? Any paper references, especially for axon degeneration stuff, would be great.
I have used ArchT-GFP CRE-reporter mice from Jax, which works well because it is inserted into the membrane. Are you transfecting or looking for transgenic lines? You just have to make sure you have a promoter that isn’t downregulated during degeneration, which can be a challenge. Maybe ATF3-CRE? Myristoylated GFP should also work well if you don’t want to put channels in there. There are plenty of membrane-targeted AAV constructs available these days, as well as some reporter mice available from Jax for that if you want to avoid using viral vectors.
Yes, that’s the one I’m using. I haven’t used the Mapt mice but I have seen some beautiful images from them during development. I would think that would work well as long as you don’t need to distinguish between sensory and sympathetic neurons in vivo.
Revisiting this topic beause I still don’t have a good solution.
Great discussion on Twitter.
I’m going to try Archon-GFP virus since it shouldn’t be affected by blue light (visualization)
