In this paper, research on the temporal and spatial distribution of cholinergic-related molecules in the lower auditory brainstem, with an emphasis on nicotinic acetylcholine receptors (nAChRs), is reviewed. The possible functions of acetylcholine (ACh) in driving selective auditory neurons before the onset of hearing, inducing glutamate receptor gene expression, synaptogenesis, differentiation, and cell survival are discussed. Experiments conducted in other neuronal and non-neuronal systems are drawn on extensively to discuss putative functions of ACh and nAChRs. Data from other systems may provide insight into the functions of ACh and nAChRs in auditory processing. The mismatch of presynaptic and postsynaptic markers and novel endogenous agonists of nAChRs are discussed in the context of non-classical interneuronal communication. The molecular mechanism that may underlie the many functions of ACh and its agonists is the regulation of intracellular calcium through nAChRs. The possible reorganization that may take place in the auditory system by the exposure to nicotine during critical developmental periods is also briefly considered.