Interferon-alpha (IFN-alpha) and transforming growth factor-beta 1 (TGF-beta 1) have been reported in different brain regions. The amygdala contains high levels of corticotropin releasing factor (CRF) and has been implicated as a central site for its stress-related autonomic and behavioral response. IFN-alpha will release arginine vasopressin (AVP) from both amygdala and hypothalamus, which further supports a role for the amygdala in neuroimmune interactions. In the present study, we compared the effects of these cytokines on the in vitro release of CRF from the amygdala and hypothalamus. In addition, we evaluated the possible involvement of guanylate cyclase-mediated signaling in CRF release. IFN-alpha stimulates CRF release from both amygdala and hypothalamus. The CRF release by IFN-alpha, Interleukin-2 (IL-2) and acetylcholine is blocked by guanylate cyclase inhibitors, indicating a role for cGMP accumulation in this CRF release. TGF-beta 1 had no effect on basal release of CRF, nor on the CRF-release induced by IL-2, but selectively blocked the acetylcholine-induced release in both amygdala and hypothalamus. Taken with a previous report that TGF-beta 1 specifically inhibits AVP release by acetylcholine, these results suggest that TGF-beta 1 may modulate HPA axis activation, by antagonizing (acetylcholine-evoked) CRF and AVP release. These data further support a role for the amygdala in the bidirectional communication between neuroendocrine and immune system.