Single or repeated mild stress increases synthesis and release of hypothalamic corticotropin-releasing factor

The effect of a specific mild stress on the levels of corticotropin-releasing factor immunoreactivity (CRF-ir) in the hypothalamus of adult male rats was determined using a radioimmunoassay specific for rat CRF. A single 5 min restraint significantly increased CRF-ir in the median eminence 24 h later compared to appropriate controls (P less than 0.025), with no change detected earlier. Plasma ACTH, an indirect index of CRF release, was significantly elevated within 15 min (P less than 0.025). Repetition of a mild stress daily for 9 days (P less than 0.01), or a single episode of handling (P less than 0.05), both resulted in significantly increased CRF-ir in the whole hypothalamus 24 h later. Blockade of axonal transport by intracisternal colchicine decreased CRF-ir in the median eminence 24 h later (P less than 0.005). Inhibition of protein synthesis by anisomycin during a single 5 minute restraint resulted in significantly decreased CRF-ir in the median eminence 24 h later compared to vehicle-injected stressed rats (P less than 0.005) or to anisomycin-injected unstressed controls (P less than 0.025). These data show that mild stress increased net hypothalamic CRF content as a result of the balance between augmented synthesis and augmented release.