The present study was aimed at measuring the distribution of ethylcholine aziridinium (AF64A)-induced cholinotoxicity within the hippocampus 6 days after bilateral (icv) administration of 1, 2 or 3 nmol, or vehicle. The dissected hippocampus was sectioned with a vibratome into 5 parallel sectors distributed along its long axis from its thalamic surface (medial) to its cortical surface (lateral). In vehicle-treated rats, the high affinity cholinergic transport (HAChT), choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activities were distributed according to a gradient of increasing activity, extending from the lateral to the medial surface of the hippocampus. After treatment with AF64A, the normal gradient of enzyme activity was profoundly disrupted at all doses of AF64A and the core sectors of the hippocampus were significantly more affected than the superficial sectors. The HAChT gradient was progressively abolished with increasing doses of toxin, and the effect was maximal at 2 nmol.