LOCALIZATION OF ATRIAL-NATRIURETIC-PEPTIDE MESSENGER-RNA AND IMMUNOREACTIVITY IN THE RAT-HEART AND HUMAN ATRIAL APPENDAGE
LOCALIZATION OF ATRIAL-NATRIURETIC-PEPTIDE MESSENGER-RNA AND IMMUNOREACTIVITY IN THE RAT-HEART AND HUMAN ATRIAL APPENDAGE
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DOI：
10.1073/pnas.84.19.6760
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发表时间：
1987-10-01
影响因子：
11.1
通讯作者：
POLAK, JM
中科院分区：
文献类型：
--
作者：
HAMID, Q;WHARTON, J;POLAK, JM
使用 32P 和 35S 标记 RNA 探针原位杂交技术，在大鼠心脏的组织切片和培养物以及人右心耳切片中研究了编码前心房钠尿肽的 mRNA 的定位。在组织切片和新生大鼠心脏培养物中，大量心房肌细胞和心室肌细胞中均显示出大鼠心房钠尿肽（ANP）转录本。这些发现与通过大鼠心脏总RNA的RNA印迹分析获得的结果一致，表明.apprxeq的单个前原-ANP转录本。心室和心房中存在 900 个核苷酸。使用 35S 标记的 RNA 探针检测人 ANP mRNA，ANP 转录本也定位于人右心耳中的大多数肌细胞。当使用与大鼠或人 ANP mRNA 编码序列相同的 RNA 探针时，仅获得放射自显影标记的背景水平。在这些制剂中，ANP 免疫反应性分布与前 ANP 前体 mRNA 之间存在密切相关性。这些结果为 ANP 基因在大鼠心室和心房中的表达提供了明确的证据，因为这些组织中的肌细胞已被确定为 ANP 定位和前体生物合成的场所。心脏培养和原位杂交的联合使用可能对未来研究心肌细胞中 ANP 合成的调节具有重要意义。
The localization of mRNA encoding preproatrial natriuretic peptide was investigated in tissue sections and cultures of rat heart and in sections of human right atrial appendage using the technique of in situ hybridization with 32P- and 35S-labeled RNA probes. Rat atrial natriuretic peptide (ANP) transcripts were demonstrated in numerous atrial myocytes and, to a lesser extent, in ventricular myocytes in both tissue sections and newborn rat heart cultures. These findings are consistent with those obtained by RNA blot analysis of rat heart total RNA, indicating that a single prepro-ANP transcript of .apprxeq. 900 nucleotides was present in the ventricles as well as the atria. Using a 35S-labeled RNA probe for human ANP mRNA, ANP transcripts were also localized to the majority of myocytes in the human right atrial appendage. Only background levels of autoradiographic labeling were obtained when RNA probes identical to the coding sequence of rat or human ANP mRNA were used. A close correlation was found between the distribution of ANP immunoreactivity and prepro-ANP mRNA in these preparations. These results provide unequivocal evidence for the expression of the ANP gene in the rat ventricles, as well as the atria, because myocytes in these tissues have been established as the sites of both ANP localization and precursor biosynthesis. The combined use of cardiac cultures and in situ hybridization may be of value in future studies investigating the regulation of ANP synthesis in cardiac myocytes.