, 1992). Lesions of the central nucleus of the amygdala that substantially diminish CRF innervation of the LC and peri-LC region have little effect on enkephalin innervation of the LC (Tjoumakaris et al., 2003). Moreover, few (2%) LC-projecting paraventricular hypothalamic nucleus neurons are enkephalin-containing, whereas 30% are immunoreactive for CRF (Reyes et al., 2005). Together these findings suggest that enkephalin and CRF axon terminals that converge onto LC neurons derive from different sources. Opioids acting at MOR on LC neurons have effects that are directly opposite to those

of CRF1 activation. MOR activation inhibits the formation of cyclic AMP and hyperpolarizes LC neurons through an increase in potassium conductance (Williams MLN2238 solubility dmso and North, 1984 and Aghajanian and Wang, 1987). In vivo MOR agonists bias LC activity towards a phasic mode, increasing synchrony and decreasing tonic discharge rate without changing or slightly increasing phasic evoked responses (Valentino and Cisplatin Wehby, 1988b and Zhu and Zhou, 2001). Like CRF, opioids

do not tonically regulate LC activity because neither MOR antagonists nor κ-opioid antagonists affect LC activity of unstressed rats (Chaijale et al., 2013, Curtis et al., 2001 and Kreibich et al., 2008). The initial evidence for stress-induced opioid regulation of LC activity came from the demonstration that systemic administration of the opioid antagonist, naloxone increased LC discharge rates of cats undergoing restraint stress, but not control cats (Abercrombie and Jacobs, 1988). Later studies using exposure to predator odor as a stress, provided evidence for CRF and enkephalin co-release during stress (Curtis et al., 2012). During this stress LC neurons shifted from a phasic to a high tonic mode, such that spontaneous discharge increased and LC and auditory-evoked discharge decreased. Administration of a CRF antagonist prior to the stress changed this response to a large inhibition of tonic

activity with slightly increased auditory-evoked activity, reminiscent of the effects of morphine administration and this was prevented by prior naloxone administration. Thus, in the presence of a CRF antagonist, exposure to the stressor tuclazepam unmasked an opioid inhibition, suggesting that both CRF and enkephalin were co-released during the stress to regulate LC discharge rate. Notably, removal of both the CRF and opioid influence in the LC by prior administration of both a CRF antagonist and naloxone rendered these neurons completely unresponsive to stressors suggesting that these afferents are the primary regulators of LC activity during acute stress (Curtis et al., 2012). CRF and opioid regulation of LC activity was also demonstrated during a physiological stressor, hypotensive stress, although the temporal aspects of opioid release during this stress were less clear (Valentino et al., 1991 and Curtis et al., 2001).