Best transgenic fluoroscent protein for visualizing axons in DRG neurons in culture

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.

@adgaudet @tberta @smithtp

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.

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@dmolliver Thanks so much! Ai32 (ChR2) similarly looks great but I wanted to avoid activating ChR2 while imaging. ArchT-GFP is a good idea then.

Is this what you’re using?

I’d prefer a transgenic mouse over virus, but could do either. I may go the ArchT route. Great idea.

Do you have any thoughts on tauGFP? This looks like a myrGFP mouse and I’ve seen it used.

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.

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TauGFP (Mapt) looks pretty good in these corneal sensory afferents from the trigeminal

Will give it a try and report back.

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)