The proliferative compartment of the intestinal crypt is critical in the process of intestinal epithelial cell homeostasis. The ability of these progenitor crypt cells to resist apoptosis and ensure restitution during a potentially lethal insult, but retain the ability to remove damaged or altered cells afterward, is necessary for preservation of the crypt-villus unit. We have examined the ability of cAMP to transiently inhibit apoptosis via the extracellular signal-regulated kinases 1 and 2 (ERK1/2), in T84 cells, an intestinal crypt-like cell line. Using the cAMP analog 8-bromo-cAMP and cholera toxin (CT), cAMP-mediated ERK1/2 activation was first measured by Western blot analysis of the phosphorylated (activated) and total (activated and inactivated) forms of ERK1/2. Cyclic AMP activated ERK1/2 in a time- and dose-dependent manner, and the effect was inhibited by PD098059, an inhibitor of the ERK1/2 signaling pathway. However, inhibition of protein kinase A (PKA) did not alter the activation of ERK1/2. CT transiently inhibited both staurosporine and Fas antibody mediated apoptosis as measured by a caspase-3 activation assay and the detection of nucleosomes in an apoptosis based enzyme-linked immunosorbent assay. This inhibitory effect was reversed by the simultaneous addition of PD098059. Our data suggest that in the T84 cell line, cAMP activates ERK1/2 in a PKA independent fashion and a physiological consequence of this activated pathway is the transient inhibition of apoptosis. These findings suggest a novel pathway that intestinal cells use to protect against injury while maintaining the overall ability to remove damaged cells and preserve intestinal homeostasis.