Age and hormonal dependence of tonin levels in rat submandibular gland as determined by a new direct radioimmunoassay.
Age and hormonal dependence of tonin levels in rat submandibular gland as determined by a new direct radioimmunoassay.
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通过新的直接放射免疫测定确定大鼠颌下腺色调水平的年龄和激素依赖性。
DOI：
10.1042/bj2380145
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Chao,J
中科院分区：
文献类型：
--
作者：
Shih,HC;Chao,L;Chao,J
建立了一种简便、灵敏的托宁直接放射免疫分析方法(EC.3.4.99)。该分析结合了一种改进的和方便的聚乙二醇法分离游离和结合的托宁。用终稀释度为1：160,000的兔抗血清，用乳过氧化物酶标记法对提纯的内毒素进行~(125)I标记。它每管检测到20pg的免疫活性托宁。大鼠颌下腺提取液系列稀释度与托宁标准曲线完全平行。与大鼠组织激肽释放酶无交叉反应。批内和批间误差分别为3.2%和5.6%。用这种方法，大鼠出生后3周(体重约50-60克)的颌下腺中就检测到了免疫反应阳性的托宁。扁桃体激素水平与年龄和性别有关，雄性大鼠的水平明显高于雌性大鼠。去势后大鼠的内分泌水平下降，17α-甲基睾酮的替代使其水平回升至高于假手术对照组。皮质醇治疗可增加去势大鼠颌下腺中的托宁水平，但甲状腺激素或雌二醇对其无影响。这种新开发的放射免疫分析方法现在可以用来测量各种组织和体液中低水平的托宁，以解决有关其调节和功能意义的问题。
A simple and sensitive direct radioimmunoassay for tonin (EC.3.4.99.-) has been developed. This assay incorporates a modified and convenient poly(ethylene glycol) technique for separation of free from bound tonin. A rabbit antiserum in a final dilution of 1:160,000 was used and the purified tonin was labelled with 125I by using a lactoperoxidase method. It detects 20 pg of immunoreactive tonin per tube. Serial dilutions of rat submandibular gland extracts showed complete parallelism with tonin standard curves. No cross-reactivity with rat tissue kallikrein was seen. Intra- and inter-assay errors were 3.2 and 5.6%, respectively. Using this assay, immunoreactive tonin was detected in the rat submandibular gland as early as 3 weeks after birth (body wt. approximately 50-60 g). Tonin levels are shown to be dependent on age and sex with significantly higher levels in male than in female rats. Castration results in decrease of tonin levels and 17 alpha-methyltestosterone replacement reversed the level to higher than the sham-operated control rats. Cortisol treatment increased, but thyroxine or oestradiol had no effect, on tonin levels in the submandibular gland of castrated rats. This newly developed radioimmunoassay can now be used to measure low levels of tonin in various tissues and body fluids to address questions about its regulation and functional significance.