At present I am not aware of the availability of any transgenic mice expressing a red flurophore under the control of a microglial specific promoter. Currently, mice available from Jackson (CD11b and CX3CR1) are only available in eGFP and YFP. A red microglial specific mouse would be great for combining with fluro transgenics (as most are eGFP) for a whole host of in-vivo imaging projects. I am strongly considering getting a line made but I was wondering whether the community was aware of any before I started moving down that pathway.
You could make a cross between a Rosa26 Td Tomato mouse with a lox-stop-lox (Jax stock number 007909) and a glial-specific Cre transgenic mouse (not sure which is most appropriate for this, maybe Jax 004600). The Cre recombinase will delete the stop codon, and allow Td Tomato to be expressed in the tissues of interest. There are likely other fluorescent proteins that are driven in this manner, if TdTomato doesn't work for your purposes.
Jackson was going to be my suggestion to check. But if there isn't one there, then I have no idea...I've not heard of one, nor have my labmates that work in this field.
You could make a cross between a Rosa26 Td Tomato mouse with a lox-stop-lox (Jax stock number 007909) and a glial-specific Cre transgenic mouse (not sure which is most appropriate for this, maybe Jax 004600). The Cre recombinase will delete the stop codon, and allow Td Tomato to be expressed in the tissues of interest. There are likely other fluorescent proteins that are driven in this manner, if TdTomato doesn't work for your purposes.
No, but there are mice strains with EGFP and EYFP expressed from the fractalkine receptor (CX3CR1) locus that label microglia very well.
Dear Phillip,
Thanks for the suggestion. Using established lines is probably a better way to go than embryo transfer. I'm fairly sure that you can get a Cre-CD11b mouse commercially. Would you recommend that Jackson, or another company do the crossing and testing?
You can do the cross yourself, it's a simple matter of breeding the two strains and using the F1 progeny. Most of the transgenic Cre mice available from Jax are sold as homozygotes (and so is the Rosa26 TdTomato), so you may not even need to genotype them. I have never had the Jackson lab do any breeding for me, in this case it's probably unnecessary.
Thanks Steven. The problem that I anticipate having is that the vast majority of other readily available commercially available lines are also 'green' or 'yellow'. Further, the green-yellow distinction is itself a bit misleading because the emission spectra for GFP and YFP are so close as to be effectively impossible completely separate. There have definitely been a couple of papers reporting crosses involving different promoters genes driving GFP and YFP. However, when I contacted several of the authors they've indicated that there is quite a lot of 'post-processing' to extract one signal from another. The reason that I'm thinking about these issues is that if I wanted to look cell-cell contacts or co-localisation than spectral bleed through becomes a real drama!
You could also try using the Runx1-MERCreMER mouse with a floxed stop reporter mouse (see http://pluchino.brc.cam.ac.uk/labwiki/images/9/9f/ginhoux_et_al._science_10.pdf)
As for the reporter, I like these mice from Jackson (Ai14: 007908 or 007914), the tdTomato is very bright and the cre efficiency is the best I have seen.
Apparently there is also a CxcR3-Cre mouse, that you could breed with the tdTomato reporter line: http://www.ncbi.nlm.nih.gov/pubmed/23273845?dopt=Abstract
It's a long shot but there are FoxP3:RFP mice (Jackson 008374). FoxP3 has been reported to be upregulated following microglial activation and acts as a repressor [of microglial actovation see PMID:20544860]. This would in theory label activated microglia...
Dr. Ransohoff at the Cleveland Clinic has mice in which the the microglia are one color and the monocytes are another but I can't remember which one is red and which is green.
There are red -green mice, CX3CR1GFP/+ (microglia) CCR2RFP/+ (macrophage). However, the microglia are green. Good luck with your plan and please let us know what you wind up doing.
Thanks Bobbi. Now that I'm getting the bills through for our new Transgenic lines I'm starting to reconsider my need for the red microglia! The paper looks really interesting. Many thanks, Rohan.
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