Microglia are resident phagocytic cells in the central nervous system (CNS), and can be activated in response to various stimuli including neurotransmitters. Using fura-2 imaging, we investigated the effects of carbachol (CCh), a cholinergic agonist, on [Ca2+]i in cultured human microglia. Treatment of microglia with CCh (100 microM) produced a transient increase in [Ca2+]i, which was atropine-sensitive and was associated with release from intracellular Ca2+ stores. Successive applications of CCh showed a change in the amplitude of the [Ca2+]i signal consistent with desensitization. These results show that human microglia express functional muscarinic receptors and respond to cholinergic agonists. The rapid change of [Ca2+]i in microglia may serve as a second messenger to trigger downstream cascades which contribute to signalling pathways in CNS pathology.