Effect of cell density on intracellular levels of the Alzheimer's amyloid precursor protein.

The precise function of APP (Alzheimer's amyloid precursor protein) remains to be fully elucidated, but various lines of evidence suggest that it may be involved in cell adhesion processes. Because APP is a transmembrane glycoprotein, variations in its expression level may have direct bearing on its putative role in cell adhesion. Our results revealed that although APP levels did not change markedly with increasing cell density (ICD), there was a small but reproducible increase in APP expression at subconfluent conditions. Higher expression APP levels led to corresponding increases in the amount of APP processed and secreted APP (sAPP) released into the cell media. Given that phorbol esters stimulate the non-amyloidogenic pathway at the expense of reducing production of Abeta (the peptide found deposited as neuritic plaques in the brains of patients with Alzheimer's disease), thus providing an interesting therapeutic focus, we tested the effect of the phorbol 12-myristate 13-acetate (PMA) on APP processing at ICD. PMA not only stimulated sAPP release at all densities tested, but also produced a corresponding decrease in the intracellular levels of APP. Further experimentation revealed that increased APP expression with ICD was dependent on factors present in conditioned medium. Interestingly, exposing cells to the Abeta peptide itself could mimic these results, thus providing evidence for a potential positive feedback mechanism between Abeta production and intracellular APP levels.