Acetylcholine is said to be involved in the neuronal transmission of photic information in the suprachiasmatic nucleus (SCN). The purpose of this study was to specify what subtypes of acetylcholine receptors play a major role using in vivo system and to examine the interaction of acetylcholine and excitatory amino acid receptors. To evaluate the effect of a selective agonist or antagonist for the nicotinic or muscarinic receptor on the neuronal transmission in the SCNT, N-acetyltransferase (NAT) activity in the pineal gland was measured after microinjection at this site. In the case of pretreatment with an antagonist, light stimulation was given after 20 min. Carbamilcholine chloride (carbachol) mimicked and only alpha-bungarotoxin (alpha-BTX) blocked the light effect; however, more selective agonists or antagonists were not effective. As for the interaction of these two cholinergic agents with an agonist or antagonist for N-methyl-D-aspartate (NMDA) receptor, alpha-BTX or D-2-amino-5-phosphonovalerate (D-APV) significantly blocked the suppressive effect of NMDA or carbachol, respectively. These data suggest that classical acetylcholine receptors do not play a direct role in neuronal transmission of photic information in the SCN in rats.