Hi all

I have been searching the net thouroghly to get ideas on how to measure the region-specific camp production in tissue slices. But everywhere it seems that all such methods ends up with camp measurements by RIA on homogenates. I was hopping some of u would know how to do this or at least have some ideas?

I used to run cAMP in cross-chopped hippocampal slices. For methods see Neuropharmacol. 2003 Vol 45, 885-894. Ultimately you do have to boil and sonicate the slices in order to relase the cAMP for RIA though. Hope that helps.

prelabeling the slices with 3Hadenine works well (the 'Shimizu' technique). You then use dowex and alumina columns to isolate 3HATP and 3HcAMP fractions, measuring cAMP production as a percent conversion from labeled ATP stores.

It's pretty simple.