The interaction between the stress axis and endogenous opioid systems has gained substantial attention as it is increasingly recognized that stress alters individual sensitivity to opiates. One site at which opiates and stress substrates may interact to have global effects on behavior is within the locus coeruleus (LC). We have previously described interactions of several opioid peptides (e.g. pro-opiomelanocortin, enkephalin) with the stress-related peptide, corticotropin-releasing factor (CRF) in the LC. To further examine interactions between DYN, ENK and CRF in the LC, sections were processed for detection of DYN, CRF or DYN and ENK in rat brain. DYN-and CRF-containing axon terminals overlapped noradrenergic dendrites in this region. Dual immunoelectron microscopy showed coexistence of DYN and CRF with 35% of axon terminals containing DYN that were also immunoreactive for CRF. In contrast, few axon terminals contained both DYN and ENK. A potential DYN/CRF afferent is the central nucleus of the amygdala (CeA). Dual in situ hybridization showed that in CeA neurons, 31% of DYN mRNApositive cells co-localized with CRF mRNA while 53% of CRF mRNAcontaining cells colocalized with DYN mRNA. Finally, to determine whether limbic DYN afferents target the LC, the CeA was electrolytically lesioned. Light-level densitometry of DYN labeling in the LC showed a significant decrease in immunoreactivity on the side of the lesion. Taken together, these data indicate that DYN-and CRF-labeled axon terminals, most likely arising from amygdalar sources are positioned to dually impact LC function whereas DYN and ENK function in parallel.