Muscarinic and purinergic receptors expressed in keratinocytes are an important part of a functional system for cell growth. While several aspects of this process are clearly dependent on Ca(2+) homeostasis, less is known about the mechanisms controlling Ca(2+) entry during epidermal receptor stimulation. We used patch-clamp technique to study responses to carbachol (CCh) and adenosine triphosphate (ATP) in HaCaT human keratinocytes. Both agonists induced large currents mediated by cation-selective channels about three times more permeable to Ca(2+) than Na(+), suggesting that they play an important role in receptor-operated Ca(2+) entry. CCh- and ATP-induced currents were inhibited by 1-[6-([(17beta)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino)hexyl]-1H-pyrrole-2,5-dione, a phospholipase C (PLC) blocker. Investigation of the pathways downstream of PLC activation revealed that InsP(3) did not affect the agonist responses. In contrast, 1-oleoyl-2-acetyl-sn-glycerol (OAG), a membrane-permeable analog of 1,2-diacylglycerol (DAG), evoked a similar cation current. This action appears to be direct, since the effects of activators or inhibitors of protein kinase C were comparatively small. Finally, transient receptor potential canonical 7 (TRPC7) specific knockdown by antisense oligonucleotides led to a decrease in ATP- and CCh-induced calcium entry, as well as OAG-evoked current. We concluded that activation of both muscarinic and purinergic receptors via a common DAG-dependent link opens Ca(2+)-permeable TRPC7 channels.