Keefer Donald A
Department of Anatomy, University of Virginia Medical School, Charlottesville, Virginia 22908.
J Histochem Cytochem. 1981 Jan;29(1A_suppl):167-174. doi: 10.1177/29.1A_SUPPL.7288152.
A refined version of the combined dry-mount auto-radiographic-immunocytochemical technique (Keefer DA, Stumpf WE, Petrusz P: Cell Tissue Res 166:25, 1976) is described in detail. In vivo nuclear estrogen uptake is measured by silver grain counting in immunocytochemically stained gonadotropes (G), somatotropes (S), lactotropes (L), corticotropes (C), and thyrotropes (T) of male rats. In rats 1 day after orchidectomy and adrenalectomy, the order of nuclear estrogen uptake was S = L G = C T, with T concentrating less than half as much radioactivity as L or S. Fifteen percent of anterior pituitary cells neither concentrated estrogen nor stained immunocytochemically. Estrogen uptake was examined in gonadotropes 1, 14, and 50 days after orchidectomy and was found to be identical at all three times. Estrogen uptake in gonadotropes of rats 14 days following orchidectomy and treatment with progesterone was reduced significantly. Guidelines for reliable quantification of the autoradiographic data is discussed.
本文详细描述了联合干片放射自显影 - 免疫细胞化学技术（Keefer DA，Stumpf WE，Petrusz P：《细胞与组织研究》166：25，1976）的改良版本。通过对雄性大鼠免疫细胞化学染色的促性腺激素细胞（G）、生长激素细胞（S）、催乳激素细胞（L）、促肾上腺皮质激素细胞（C）和促甲状腺激素细胞（T）进行银颗粒计数，来测量体内细胞核对雌激素的摄取。在去势和肾上腺切除术后1天的大鼠中，细胞核摄取雌激素的顺序为S = L＞G = C＞T，其中T聚集的放射性不到L或S的一半。垂体前叶细胞中有15%既不聚集雌激素也不被免疫细胞化学染色。对去势后1天、14天和50天的促性腺激素细胞中的雌激素摄取进行了检测，发现这三个时间点的摄取情况相同。去势14天后并用孕酮处理的大鼠促性腺激素细胞中的雌激素摄取显著降低。讨论了放射自显影数据可靠定量的指导原则。
