In situ hybridization for creatine kinase-B messenger RNA in rat uterus and brain.
In situ hybridization for creatine kinase-B messenger RNA in rat uterus and brain.
复制标题
大鼠子宫和大脑中肌酸激酶 B 信使 RNA 的原位杂交。
DOI：
10.1016/0303-7207(93)90081-t
复制
发表时间：
1993
影响因子：
4.1
通讯作者：
Pfaff,DW
中科院分区：
文献类型：
--
作者：
Bergen,HT;Pentecost,BT;Dickerman,HW;Pfaff,DW
肌酸激酶-B (CKB) 存在于子宫和大脑中，在子宫中其合成（蛋白质和 mRNA）受雌激素调节。在本研究中，我们采用原位杂交技术检测子宫和大脑中的CKB mRNA，并确定这些组织中是否存在雌激素对CKB的细胞类型特异性诱导。组织取自卵巢切除 (ovx) 大鼠，这些大鼠在 2、8、24 和 72 小时前注射了雌激素（17β-雌二醇-3-苯甲酸酯和/或 17β-雌二醇）或单独的载体。取出大脑和子宫，冷冻，低温切片，并进行原位杂交组织化学处理。将子宫和脑切片与与 CKB mRNA 3' 片段互补的氚化 DNA 探针或与相同 3' 片段互补的对照正义探针一起孵育。在子宫平滑肌细胞中，在雌激素给药后 2 小时和 24 小时观察到 CKB mRNA 的诱导分别增加了 2.5 倍和 3.5 倍，并且水平在 72 小时时接近 ovx 对照。在子宫上皮中观察到较小的诱导（1.9 倍，2 小时），在基质中几乎没有 CKB mRNA 的诱导。在大脑中，CKB mRNA 在神经元中检测到，但在明确识别的神经胶质细胞中未检测到，仅偶尔在室管膜细胞中检测到。在含有雌激素受体的大脑区域中，没有证据表明雌激素对 CKB mRNA 水平有显着影响。一些大脑区域的神经元表达高于其他区域（例如内侧隔膜与区域），但表达广泛且不限于神经内分泌位点。
Creatine kinase-B (CKB) is present in both uterus and brain, and in uterus its synthesis (protein and mRNA) is regulated by estrogen. In the present study we have used in situ hybridization to detect CKB mRNA in uterus and brain, and to determine whether there is cell type specific induction of CKB by estrogen in these tissues. Tissue was taken from ovariectomized (ovx) rats that, had been injected with either estrogen (17β-estradiol-3-benzoate and/or 17β-estradiol) or vehicle alone, 2, 8, 24 and 72 h previously. The brains and uteri were removed, frozen, cryostat-sectioned, and processed for in situ hybridization histochemistry. The uterine and brain sections were incubated with a tritiated DNA probe complementary to a 3' fragment of CKB mRNA, or a control sense probe to the same 3' fragment. In uterine smooth muscle cells, a 2.5- and 3.5-fold induction of CKB mRNA was observed 2 and 24 h after estrogen administration, respectively, and levels approached ovx controls at 72 h. A smaller induction (1.9-fold, 2 h) was observed in uterine epithelium, with little induction of CKB mRNA in stroma. In the brain CKB mRNA was detected in neurons, but not in clearly identified glia, and only occasionally in ependymal cells. In brain regions containing estrogen receptors there was no evidence of a significant estrogen effect on CKB mRNA levels. Some brain regions had higher neuronal expression than others (e.g. medial septum vs. area), but expression was widespread and not limited to neuroendocrine sites.