... according to the manufacturer’s protocol.Total RNA (1 lg) was reverse-transcribed to obtain single-stranded cDNA, using a kit (Toyobo, Tokyo, Japan). Spe-cific primers for human cholinergic receptors ... acetylcho-linesterase inhibitor physostigmine was obtained fromWako Pure Chemical Industries (Osaka, Japan). Donepezil,another acetylcholinesterase inhibitor, was a generous giftfrom Eisai (Tokyo, ... microspheres of human serum albumin withperflutren for contrast enhancement during ultrasoundimaging (Optison; GE Healthcare, Princeton, NJ, USA).The vectors were injected into the left quadriceps...