ESTROGEN RECEPTOR-IMMUNOSTAINING OF NEURONAL CYTOPLASMIC PROCESSES AS WELL AS CELL-NUCLEI IN GUINEA-PIG BRAIN
ESTROGEN RECEPTOR-IMMUNOSTAINING OF NEURONAL CYTOPLASMIC PROCESSES AS WELL AS CELL-NUCLEI IN GUINEA-PIG BRAIN
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DOI：
10.1016/0006-8993(89)91219-5
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发表时间：
1989-08-21
期刊：
影响因子：
2.9
通讯作者：
TURCOTTE, JC
中科院分区：
文献类型：
--
作者：
BLAUSTEIN, JD;TURCOTTE, JC
我们最近开发了一种免疫细胞化学技术，用于在豚鼠大脑中的一些细胞的细胞核中染色雌激素受体。通过对这一技术的优化，我们现在已经成功地对豚鼠脑内许多神经元的突起进行了雌激素受体免疫反应的染色，这些神经元也含有雌激素受体免疫反应细胞核。虽然在各种条件下，反应产物在细胞核中都可见，但神经元突起只有在对程序进行优化以获得最高灵敏度的情况下才会被深色免疫染色，例如，多桥免疫细胞化学法。去掉Triton X-100和/或二甲基亚砜，通常用来增加抗体的穿透性，对突起或细胞核的免疫染色没有影响，在振动切片和冷冻切片上都有明显的免疫染色。注射雌二醇，但不注射黄体酮，导致胞浆雌激素受体免疫染色完全消失，这与胞浆免疫染色与细胞核免疫染色一样是由于雌激素受体的存在而引起的观点是一致的。在所有含有雌激素受体免疫反应细胞核的神经解剖区域，部分(但不是全部)细胞的相关突起也被免疫染色。然而，在某些区域，如中脑中央灰质和视前区，细胞质染色特别深。导致一些雌激素受体免疫反应阳性细胞免疫染色的细胞特征正在调查中。
We have recently developed an immunocytochemical technique for staining estrogen receptors in cell nuclei of some cells in the guinea pig brain. With optimization of this technique, we have now been successful in staining estrogen receptor-immunoreactiivty in processes of many neurons in the guinea pig brain that also contain estrogen receptor-immunoreactive cell nuclei. While reaction product is visible in cell nuclei under a wide variety of conditions, neuronal processes are darkly immunostained only with modifications of the procedure optimized for maximum sensitivity, for example, the multiple-bridge immunocytochemical procedure. Omission of Triton X-100 and/or dimethysulfoxide, usually used to increase penetration of the antibodies, had no effect on immunostaining in processes or cell nuclei, and immunostaining was apparent in both vibratome-cut and freezing microtome-cut sections. Injection of estradiol, but not progesterone, caused the total loss of cytoplasmic estrogen receptor-immunostaining, consistent with the idea that the cytoplasmic immunostaining, like the cell nuclear immunostaining is due to the presence of estrogen receptors. In all neuroanatomical regions containing estrogen receptor-immunoreactive cell nuclei, associated processes of some, but not all cells were also immunostained. However, in certain areas, such as the midbrain central gray and the preoptic area, cytoplasmic staining was particularly dark. The cellular characteristics that result in immunostaining in some estrogen receptor-immunoreactive cells, and not in others is under investigation.