Epidermal Growth Factor Modulation of Corticotrope Function

本研究的目的是确定生长因子（EGF）的来源和靶细胞。此外，由于它已被证明可以刺激垂体前叶促肾上腺皮质激素，目的是确定垂体egf产生细胞是否对激活促肾上腺皮质激素分泌的相同刺激作出反应，即应激和促肾上腺皮质激素释放激素（CRH）。为了确定EGF的来源，将信使核糖核酸（mRNA）定位到EGF的标记协议将被完成。源细胞将通过垂体激素的双重标记进一步鉴定。一旦确定源细胞，将确定它们对应激或CRH刺激的反应，以了解它们是否可以通过增加EGF mRNA来激活。以EGF靶细胞为目标。这些细胞上的EGF受体将被标记为生物素化的EGF类似物或针对EGF受体的抗体。然后这些细胞将被进一步识别为激素的标签。然后源细胞将被富集并刺激分泌。培养基将与促肾上腺皮质激素一起使用，以确定细胞是否分泌具有生物活性的表皮生长因子。反应的特异性将通过向EGF中加入抗血清来中和任何EGF活性来确定。

The objective of this study is to determine the source and target cells for a growth factor (EGF). Furthermore, because it has been shown to stimulate anterior pituitary corticotropes, the aims will determine if pituitary EGF-producing cells respond to the same stimuli that activate secretion from the corticotropes namely stress and corticotropin releasing hormone (CRH). To identify the source of EGF, labeling protocols that localize messenger ribonucleic acid (mRNA) to EGF will be done. The source cells will then be further identified by dual labeling for pituitary hormones. Once the source cells are identified, their responses to stimulation by stress or CRH will be determined to learn if they can be activated by increasing EGF mRNA. Aim target cells for EGF. EGF receptors on these cells will be labeled either with a biotinylated analog of EGF or antibodies to the EGF receptor. Then these cells will be identified further with labels for the hormones. The source cells will then be enriched and stimulated to secrete. The media will be used with corticotropes to determine if the cells secrete biologically active EGF. The specificity of the reaction will be determined by adding antisera to EGF to neutralize any EGF activity.