Koivusalo F, Leppäluoto J, Knip M, Rajaniemi H
Acta Endocrinol (Copenh). 1981 Jul;97(3):398-404. doi: 10.1530/acta.0.0970398.
Isolated rat pancreatic islet extracts were submitted to reverse phase high pressure chromatography (HPLC) and assayed for thyrotrophin releasing factor (TRF), glucagon and insulin in radioimmunoassays (RIA). Our anti-TRF serum was prepared by immunizing rabbits with TRF conjugated to bovine thyroglobulin (bTG) by bisdiazobenzidine. Anti-glucagon and antiinsulin were from a commercial source (Novo, Denmark). The concentrations of TRF, glucagon and insulin were 0.05 +/- 0.02, 0.06 +/- 0.02 and 1.0 +/- 0.2 pmol/islet, respectively. Formalin-fixed pancreatic sections were stained for TRF, glucagon and insulin by peroxidase, antiperoxidase (PAP) complex method. When the adjacent sections were stained for glucagon or insulin, it was observed that TRF and glucagon-specific peroxidase reactions were confined to the marginal islet cells, but insulin reactions to the central cells. The TRF-specific peroxidase reaction was clearly reduced when the anti-TRF serum was unchanged when the antiserum was pre-incubated with synthetic TRF. The present HPLC results suggest that the islets contained TRF. Immunocytochemical studies show that the TRF-immunoreactive material, either synthesized or bound, is localized in the marginal islet cells.
将分离的大鼠胰岛提取物进行反相高压液相色谱（HPLC）分析，并通过放射免疫测定法（RIA）检测促甲状腺激素释放因子（TRF）、胰高血糖素和胰岛素。我们的抗TRF血清是通过用双偶氮联苯胺将TRF与牛甲状腺球蛋白（bTG）偶联后免疫兔子制备的。抗胰高血糖素和抗胰岛素抗体购自商业来源（丹麦诺和诺德公司）。TRF、胰高血糖素和胰岛素的浓度分别为0.05±0.02、0.06±0.02和1.0±0.2 pmol/胰岛。用过氧化物酶-抗过氧化物酶（PAP）复合物法对福尔马林固定的胰腺切片进行TRF、胰高血糖素和胰岛素染色。当相邻切片用胰高血糖素或胰岛素染色时，观察到TRF和胰高血糖素特异性过氧化物酶反应局限于胰岛边缘细胞，而胰岛素反应则在中央细胞。当抗血清与合成TRF预孵育时，抗TRF血清未改变时，TRF特异性过氧化物酶反应明显减弱。目前的HPLC结果表明胰岛中含有TRF。免疫细胞化学研究表明，合成或结合的TRF免疫反应性物质定位于胰岛边缘细胞。
