Microglial morphology and its transformation after challenge by extracellular ATP in vitro

The novel morphological characteristics of N9 microglial cells and primary cultured rat microglial cells were examined using the bitotin-IB4 and streptavidin-FITC system. Numerous fine, long processes of both microglial cell preparations formed a network, beginning after 30 min in culture. Dye coupling studies did not show communication between neighbouring cells via the processes in normal conditions. The network of microglial cell processes was well formed into a 'resting state' by 16-24 hr after re-plating. After being challenged by 3 mM ATP the microglial cells were activated, became amoeboid-like cells within 2 hr and finally floated in the culture medium. The complicated network of processes did not retract to the microglial cell body. Flow cytometry analysis showed that the majority of these floating cells were alive and could recover to the resting state after ATP was removed from the culture medium.