We are trying to replicate the endogenous tau found in TgF344 AD rats (Cohen et al. J Neurosci. 2013 Apr 10;33(15):6245-56. doi: 10.1523/JNEUROSCI.3672-12.2013). We have used many antibodies - CP13, AT8, MC1, PHF1 - with DAB &/or tyramide booster, and haven't found anything. Please let me know if you have tried anything that works.
Hi @Claire Galloway,
Have you tried To contact the authors of the publication you have mentioned (Cohen et al. J Neurosci. 2013)?
I would certainly give it a try first. Sometime you can't find all the details in material and methods section and contacting the authors might help.
Do you use paraffin section or cryopreservation tissue blocks?
For paraffin sections there are many different methods to improve exposure of antigen determinants. Boiling the slides in citrate buffer for 5 min, for example, is one way to enhance antigen display of denatured proteins. Such processing may increase background staining but if it works I consider it as an acceptable trade-off.
Hope this helps
Amiel
Wish I had the answer for you. I asked every TgF344-AD group I could find at SfN this question last year. Turns out many of them were also struggling to replicate the Cohen et al tau findings.
You’ve probably already seen this TgF344-AD study, but in case you haven’t, this is one of the few papers I’ve seen the literature that documents tau in these animals outside of the Cohen group.
https://www.biologicalpsychiatryjournal.com/article/S0006-3223(17)32145-5/pdf
I ended up shifting my focus to all things amyloid and neuron loss for the sake of finishing my dissertation next year. I do see amyloid beta for daaaays in my aged TgF344-AD rats with ThioS staining, and so do our offsite collabs.
Hope this helps.
Danielle E. Panoz-Brown thank you for your help! I will try to contact those authors and see if they have any suggestions.
Also Rorabaugh et al (https://academic.oup.com/brain/article/140/11/3023/4222757) found phosphorylated tau in the locus coeruleus of the TgF344 AD rats using CP13 Ser205 phospho-antibody, but we haven't had much luck with that one lately.
Hi Claire,
According to my experience pTau in such models is often not as you would expect and comparable to a human AD brain. Typically you should also find hyperphosphorylation of some neurites within neuritic plaques. What could help you is using TrueBlack as a quencher for autofluorescence and then label with MOAB2 + pThr231 Tau [ EPR2488]. This one may be less prone to conformational differences from fixation procedures, and if you are using citrate buffer steaming (if FFPE) you might be able to increase labeling results. pThr231 is typically also found naturally to some extent, so you should also know from WT that the labeling works. Beside that this p Site is early hyperphosphorylated in AD and is typically also one of the first to become positive in amyloid/PS models.
Just as idea,
Good luck,
Daniel
Thank you Daniel Havas , we will give that a try! Would you happen to have a full protocol with this technique that worked for your lab?
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