Hashimoto K, Ohno N, Yunoki S, Kageyama J, Aoki Y, Takahara J, Ofuji T
Endocrinol Jpn. 1981 Feb;28(1):1-7. doi: 10.1507/endocrj1954.28.1.
Hypothalamic median eminence extracts (MEE) were subjected to gel filtration on Sephadex G-25 and G-75 columns to characterize the corticotropin releasing factor (CRF) in relation to arginine vasopressin (AVP). CRF activity was measured using monolayer cultured anterior pituitary cells, and AVP was measured by radioimmunoassay. Sephadex G-25 chromatography of AVP immunoreactivity of MEE (NIAMDD-Rat HE-RP-1) showed three peaks on elution with 0.1 N HCI and four peaks on elution with 0.2M acetic acid. But freshly prepared MEE showed only one peak on elution with 0.1 N HCI. These results suggest that AVP in NIAMDD-Rat HE-RP-1 is polymerized or aggregated on the Sephadex G-25 column, especially when eluted in 0.2M acetic acid. Two main peaks of CRF activity appeared consistently on both Sephadex G-25 and G-75 chromatography. One was near the void volume and the other was retarded. The small molecular CRF (small-CRF) peak was coeluted with immunoreactive AVP and 125I-AVP, on both chromatographies on elution with 0.1 N HCI. The large molecular CRF (big-CRF) appeared between the void volume and I-39 ACTH on Sephadex G-75 chromatography. Big-CRF from freshly prepared MEE had no AVP immunoreactivity. AVP showed CRF activity in pituitary cell cultures, but its CRF activity accounted for no more than 20% of the CRF activity of NIAMDD-Rat HE-RP-1. The log dose-response characteristics of the CRF activities of small-CRF and AVP differed. These results suggest that the median eminence has at least two substances with CRF activity: one is large molecular CRF, and the other is small molecular CRF which may have a vasopressin-like molecular weight. AVP may account for a part of the CRF activity of small molecular CRF but is not identical with genuine CRF.
对下丘脑正中隆起提取物（MEE）进行葡聚糖G - 25和G - 75柱凝胶过滤，以鉴定促肾上腺皮质激素释放因子（CRF）与精氨酸加压素（AVP）的关系。使用单层培养的垂体前叶细胞测量CRF活性，通过放射免疫测定法测量AVP。MEE（NIAMDD - 大鼠HE - RP - 1）的AVP免疫反应性经葡聚糖G - 25色谱分析，用0.1N HCl洗脱时出现三个峰，用0.2M乙酸洗脱时出现四个峰。但新鲜制备的MEE用0.1N HCl洗脱时仅出现一个峰。这些结果表明，NIAMDD - 大鼠HE - RP - 1中的AVP在葡聚糖G - 25柱上发生了聚合或聚集，尤其是在0.2M乙酸洗脱时。在葡聚糖G - 25和G - 75色谱上均始终出现两个主要的CRF活性峰。一个靠近空体积，另一个有延迟。在0.1N HCl洗脱的两种色谱上，小分子CRF（small - CRF）峰与免疫反应性AVP和125I - AVP共洗脱。大分子CRF（big - CRF）在葡聚糖G - 75色谱的空体积和I - 39促肾上腺皮质激素之间出现。新鲜制备的MEE中的big - CRF没有AVP免疫反应性。AVP在垂体细胞培养物中显示出CRF活性，但其CRF活性不超过NIAMDD - 大鼠HE - RP - 1的CRF活性的20%。small - CRF和AVP的CRF活性的对数剂量反应特征不同。这些结果表明，正中隆起至少有两种具有CRF活性的物质：一种是大分子CRF，另一种是可能具有类似加压素分子量的小分子CRF。AVP可能占小分子CRF的部分CRF活性，但与真正的CRF不同。
