In vivo release of angiotensin II from the rat hypothalamus

Recent studies suggest that angiotensin II is released from neuronal tissue in vitro, but the occurrence of this phenomenon in the intact brain has not yet been demonstrated. To characterize the in vivo release of immunoreactive angiotensin II, push-pull cannulas were positioned in the anterior hypothalamus in 47 Sprague-Dawley rats (200-250 g) anesthetized with Inactin (100 mg/kg i.p.). Artificial cerebrospinal fluid was perfused at 20 microliters/min, and effluent samples were collected for 15-minute periods over 2 hours. Angiotensin II was detectable (greater than 2.5 pg/ml) in the push-pull cannula perfusate of the majority (76%) of the rats. Spontaneous release of immunoreactive angiotensin II was constant for 2 hours in 11 rats at values averaging from 4.4 +/- 1.5 to 8.2 +/- 2.2 pg/ml. In addition, bilateral nephrectomy performed 48 hours before did not affect the detection of angiotensin II (n = 3). Angiotensin immunoreactivity in the rat hypothalamus was further characterized by high performance liquid chromatography. The analysis showed that the perfusate contained authentic angiotensin II as well as other angiotensin metabolites. The effect of beta-adrenergic modulation on the release of angiotensin II was assessed in 20 rats by adding isoproterenol (10(-10), 10(-8), and 10(-6) M), propranolol (10(-6) M), or a combination of both. Neither activation nor inhibition of hypothalamic beta-receptors altered the spontaneous release of angiotensin II. These data demonstrate that angiotensin II and congener peptides are detectable in the microenvironment of the anterior hypothalamus of the anesthetized rat and that the release of angiotensin II immunoreactivity in the anterior hypothalamus is not modified by beta-adrenergic receptors.