It has been suggested that cholinergic effects in the rat cochlear nucleus (CN) are mediated by muscarinic acetylcholine receptors. In this study, immunohistochemistry for muscarinic subtype 2 (m2) receptors using a monoclonal subtype-specific antibody (Levey et al. [1995] J. Comp. Neurol. 351:339-356) revealed an m2-like system in the rat CN. A prominent lamina of m2-immunoreactive fibers and puncta was located in a subgranular layer of the caudal anteroventral cochlear nucleus (AVCN) and the posteroventral cochlear nucleus (PVCN). The superficial granular layer of the rostral AVCN and the medial sheet region also contained notable immunoreactivity for m2. Some labeled somata and their processes were found in magnocellular regions of the ventral CN. A network of neurites and puncta was located in the fusiform soma and deep layers of the dorsal CN. The olivocochlear bundle and its branches to the CN were also m2 immunoreactive and possibly contributed m2-labeled fibers and terminals to the CN. Some similarities and some differences were found between this m2 receptor distribution pattern and previous results for choline acetyltransferase (ChAT), acetylcholinesterase (AChE), and muscarinic acetylcholine receptor immunohistochemistry and binding in the CN. The results suggest that m2 receptors that are located both pre- and postsynaptically mediate many cholinergic effects in the rat CN.