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Historically, ACh has been implicated in learning and short-term memory functions. However, more recent studies have provided support for a role of cortical ACh in attentional effort, orienting and the detection of behavioral significant stimuli. The current review article summarizes studies in animals and humans which have investigated the role of ACh in attention and cognition. An attempt has been made to differentiate between brain regions involved in attentional processes versus those important for other cognitive functions. To this purpose, various experimental methods and interventions were used. Animal behavioral studies have injected the selective immunotoxin IgG-saporin to induce specific cholinergic lesions, employed electrochemical techniques such as microdialysis, or have administered cholinergic compounds into discrete parts of the brain. Human studies that give some indication on the link between central cholinergic signaling and cognition are obviously confined to less invasive, imaging methods such as fMRI. The brain areas that are deemed most important for intact attentional processing in both animals and humans appear to be the (pre)frontal, parietal and somatosensory (especially visual) regions, where ACh plays a vital role in the top-down control of attentional orienting and stimulus discrimination. In contrast, cholinergic signaling in the septohippocampal system is suggested to be involved in memory processes. Thus, it appears that the role of ACh in cognition is different per brain region and between nicotinic versus muscarinic receptor subtypes.