A PERIFUSION METHOD FOR EXAMINING ARGININE VASOPRESSIN (AVP) RELEASE FROM HYPOTHALAMO-NEUROHYPOPHYSEAL SYSTEM

Abstract

A perifusion method was developed using rat hypothalamo-neurohypophyseal system (HNS) or neural lobe to investigate the control mechanism of arginine vasopressin (AVP) release. A specific radioimmunoassay (RIA) for AVP was developed to measure AVP in perifusion medium employing anti-AVP serum which was obtained by immunizing rabbits. At a final dilution of 1/12,000 the antiserum showed less than 0.66 and 0.01% cross reactivity with lysine-vasopressin and oxytocin, respectively. It did not cross react with other peptide hormones. The lowest detectable level of vasopressin was 0.5 pg/tube. The intra-assay coefficient of variation averaged 10.4%. The dilution curve of perifused medium was well paralled to the standard curve of AVP assay. AVP release from HNS or neural lobe gradually declined to the stable level in 90-120 min after the initiation of perifusion. Good repeatability of the AVP release from neural lobe was recognized by repeated stimulation with 10 min perifusion of 60 mM KCl at every 60 min. HNS released AVP in dose related manner to the osmotic challenge of Na or glucose, and AVP release was stimulated from HNS by prostaglandin E2, but not by dopamine. The perifusion method using AVP-RIA is a useful method to examine the AVP release from HNS or neural lobe.