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正常膀胱中的激活素及其受体:免疫组织化学、原位杂交和逆转录聚合酶链反应

Activin and activin receptors in the normal urinary bladder: immunohistochemistry, in situ hybridization, and RT-PCR.

作者信息

Ying S Y, Zhang Z

机构信息

Department of Cell and Neurobiology, University of Southern California School of Medicine, Los Angeles 90033, USA.

出版信息

Life Sci. 1995;57(17):1599-603. doi: 10.1016/0024-3205(95)02135-6.

Abstract

To determine whether rat urinary bladder produces activin-A, a multifunctional growth factor in the transforming growth factor beta superfamily (TGF beta), we have conducted immunohistochemistry using specific antibodies for activin beta A-subunit which were raised against a synthetic cyclic fragment of the beta A-subunit of activin. The mature activin-A molecule was identified at transitional epithelial cells, smooth muscle, and endothelial cells. To determine if messenger RNAs for the beta A-subunits of activin-A and activin receptors are expressed in these cells, both in situ hybridization with specific probes and the reverse transcription-polymerase chain reaction (RT-PCR) technique with primers specific for the beta A-subunit of activin and activin receptors, respectively, were used. Messenger RNA expression of the beta A-subunit of activin-A and activin receptors were detected by RT-PCR and localized in the transitional epithelial, smooth muscle, and endothelial cells as determined by in situ hybridization. In addition, the identity of the cDNA product of RT-PCR was verified with DNA sequencing. The localization of mature activin-A protein and its corresponding message as well as that of activin receptors to urinary bladder cells suggest that activin-A may have an autocrine function in the urinary bladder, perhaps in the transitional epithelial, smooth muscle, and endothelial cells themselves.

摘要

为了确定大鼠膀胱是否产生激活素-A(一种转化生长因子β超家族中的多功能生长因子),我们使用针对激活素βA亚基的特异性抗体进行了免疫组织化学研究,这些抗体是针对激活素βA亚基的合成环状片段制备的。在移行上皮细胞、平滑肌细胞和内皮细胞中鉴定出了成熟的激活素-A分子。为了确定激活素-A的βA亚基和激活素受体的信使核糖核酸是否在这些细胞中表达,分别使用了与特异性探针的原位杂交技术以及分别针对激活素βA亚基和激活素受体的引物进行逆转录-聚合酶链反应(RT-PCR)技术。通过RT-PCR检测到激活素-A的βA亚基和激活素受体的信使核糖核酸表达,并通过原位杂交确定其定位于移行上皮细胞、平滑肌细胞和内皮细胞。此外,通过DNA测序验证了RT-PCR的cDNA产物的同一性。成熟激活素-A蛋白及其相应信使以及激活素受体在膀胱细胞中的定位表明,激活素-A可能在膀胱中具有自分泌功能,也许在移行上皮细胞、平滑肌细胞和内皮细胞自身中发挥作用。

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