Jung-Testas I, Renoir M, Bugnard H, Greene G L, Baulieu E E
Université Paris XI, Laboratoire Hormones, Bicêtre, France.
J Steroid Biochem Mol Biol. 1992 Mar;41(3-8):621-31. doi: 10.1016/0960-0760(92)90394-x.
Primary cultures of rat glial cells were established from newborn rat forebrains. A mixed population of oligodendrocytes and astrocytes was obtained, as confirmed by indirect immunofluorescence staining with specific markers for each cell type. Receptors were measured 3 weeks after primary culture in glial cells cultured in the presence or not of 50 nM estradiol and we have identified progesterone, glucocorticoid, estrogen, and androgen receptors (PR, GR, ER and AR), but only PR was inducible by the estrogen treatment. This estrogen-induction of PR was more dramatic in glial cells derived from female offsprings than from males, as measured by binding studies and by immunohistochemical techniques with the KC 146 anti-PR monoclonal antibody. The antiestrogen tamoxifen inhibited the estrogen induction, but had no effect by itself on PR concentration. Specific binding sites for PR, GR, ER and AR were measured by whole cell assays after labeling cells with, respectively, [3H]R5020, [3H]dexamethasone, [3H]OH-tamoxifen or [3H]R1881. PR and GR were also analyzed by ultracentrifugation and after exposure of cells to agonists, both receptors were recovered from cytosol as a 9S form, and from the nuclear high-salt, tungstate ions-containing fraction as a 4-6S form. In contrast, when the antiprogestin- and antiglucocorticosteroid RU486 was used as a ligand, a non-activated 8.5S receptor complex was found for both receptors in this nuclear fraction. The 8.5S complex of the GR was further analyzed in the presence of specific antibodies and, in addition to GR, the presence of the heat shock protein hsp90 and of a 59 kDa protein was found. During primary culture, the effects of progesterone (P) and estradiol (E2) were tested on glial cell multiplication, morphology and differentiation. Cell growth was inhibited by P and stimulated by E2. Both hormones induced dramatic morphologic changes in oligodendrocytes and astrocytes and increased synthesis of the myelin basic protein in oligodendrocytes and of the glial fibrillary acidic protein in astrocytes.
大鼠神经胶质细胞的原代培养物取自新生大鼠的前脑。通过用每种细胞类型的特异性标记物进行间接免疫荧光染色证实，获得了少突胶质细胞和星形胶质细胞的混合群体。在原代培养3周后，对在存在或不存在50 nM雌二醇的情况下培养的神经胶质细胞中的受体进行了检测，我们鉴定出了孕酮、糖皮质激素、雌激素和雄激素受体（PR、GR、ER和AR），但只有PR可被雌激素处理诱导。通过结合研究以及使用KC 146抗PR单克隆抗体的免疫组织化学技术测定，这种PR的雌激素诱导在源自雌性后代的神经胶质细胞中比在雄性后代的神经胶质细胞中更显著。抗雌激素他莫昔芬抑制雌激素诱导，但自身对PR浓度没有影响。分别用[3H]R5020、[3H]地塞米松、[3H]羟基他莫昔芬或[3H]R1881标记细胞后，通过全细胞测定法测量PR、GR、ER和AR的特异性结合位点。还通过超速离心以及在细胞暴露于激动剂后对PR和GR进行了分析，两种受体均以9S形式从细胞质中回收，并以4 - 6S形式从含钨酸盐离子的核高盐级分中回收。相反，当使用抗孕激素和抗糖皮质激素RU486作为配体时，在该核级分中发现两种受体均存在非活化的8.5S受体复合物。在特异性抗体存在的情况下对GR的8.5S复合物进行了进一步分析，除了GR外，还发现了热休克蛋白hsp90和一种59 kDa蛋白的存在。在原代培养期间，测试了孕酮（P）和雌二醇（E2）对神经胶质细胞增殖、形态和分化的影响。P抑制细胞生长，E2刺激细胞生长。两种激素均诱导少突胶质细胞和星形胶质细胞发生显著的形态变化，并增加少突胶质细胞中髓鞘碱性蛋白以及星形胶质细胞中胶质纤维酸性蛋白的合成。
