Ohno N, Hashimoto K, Yunoki S, Takahara J, Ofuji T
Acta Med Okayama. 1981 Feb;35(1):27-35. doi: 10.18926/AMO/31292.
A perifusion method has been 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. But it did not cross reacted 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 sodium or glucose, and AVP release was stimulated from HNS by prostaglandin E2, but not by dopamine. These results show that the perifusion methods using AVP-RIA is a useful method to examine the AVP release from HNS or neural lobe.
已开发出一种使用大鼠下丘脑 - 神经垂体系统（HNS）或神经叶的灌流方法，以研究精氨酸加压素（AVP）释放的控制机制。开发了一种针对AVP的特异性放射免疫测定法（RIA），以使用通过免疫兔子获得的抗AVP血清来测量灌流培养基中的AVP。在最终稀释度为1/12,000时，该抗血清与赖氨酸加压素和催产素的交叉反应性分别低于0.66%和0.01%。但它与其他肽类激素没有交叉反应。加压素的最低可检测水平为0.5 pg/管。测定内变异系数平均为10.4%。灌流培养基的稀释曲线与AVP测定的标准曲线良好平行。在灌流开始后90 - 120分钟内，HNS或神经叶释放的AVP逐渐下降至稳定水平。通过每60分钟用60 mM KCl进行10分钟的灌流重复刺激，可观察到神经叶释放AVP具有良好的重复性。HNS以与钠或葡萄糖的渗透压刺激相关的剂量方式释放AVP，并且前列腺素E2可刺激HNS释放AVP，但多巴胺不能。这些结果表明，使用AVP - RIA的灌流方法是检查HNS或神经叶释放AVP的有用方法。
