Herman M A, Schulz C A, Claude P
Department of Physiology, University of Wisconsin, Madison 53715.
Exp Cell Res. 1992 Jun;200(2):370-8. doi: 10.1016/0014-4827(92)90185-b.
Nerve growth factor (NGF) promotes the outgrowth of neurites from cultured adrenal chromaffin cells from adult rhesus monkeys, but little is known about the distribution, at the cellular level, of the NGF receptors (NGFR) responsible for this response. We examined changes in immunostaining for NGFR in chromaffin cells cultured for 4 weeks in the presence or absence of NGF, with or without dexamethasone (DEX), which inhibits neuritic outgrowth from these cells. Purified cultures of adrenal chromaffin cells from adult rhesus monkeys were grown for up to 9 weeks in NGF, DEX, NGF plus DEX, or control medium. Cells were immunolabeled with three different monoclonal antibodies directed against different epitopes of the human NGFR. Although the distribution of immunolabeling was not uniform from cell to cell, the overall intensity of NGFR immunolabeling varied dramatically between different growth conditions. Of greatest interest, DEX-treated cells stained the most intensely at all time points, while the intensity of immunolabeling was much fainter in NGF-treated cells and decreased with time in culture. In contrast to the intensity of labeling, the proportion of chromaffin cells with immunoreactivity increased with time in all treatment groups. Thus, GCs do not appear to antagonize the effects of NGF merely by decreasing the total number of immunoreactive NGFR on the surface of these cells.
神经生长因子(NGF)可促进成年恒河猴培养的肾上腺嗜铬细胞长出神经突,但对于介导此反应的NGF受体(NGFR)在细胞水平的分布情况却知之甚少。我们检测了在有无NGF以及有无地塞米松(DEX,其可抑制这些细胞的神经突生长)存在的情况下,培养4周的嗜铬细胞中NGFR免疫染色的变化。将成年恒河猴的肾上腺嗜铬细胞纯化培养物在NGF、DEX、NGF加DEX或对照培养基中培养长达9周。用三种针对人NGFR不同表位的不同单克隆抗体对细胞进行免疫标记。尽管细胞间免疫标记的分布并不均匀,但在不同生长条件下,NGFR免疫标记的总体强度差异很大。最令人感兴趣的是,DEX处理的细胞在所有时间点染色都最深,而NGF处理的细胞免疫标记强度要弱得多,且随着培养时间的延长而降低。与标记强度相反,所有处理组中具有免疫反应性的嗜铬细胞比例随时间增加。因此,糖皮质激素似乎并非仅仅通过减少这些细胞表面免疫反应性NGFR的总数来拮抗NGF的作用。