In situ hybridization of H-K-ATPase beta-subunit mRNA in rat and rabbit kidney.
In situ hybridization of H-K-ATPase beta-subunit mRNA in rat and rabbit kidney.
复制标题
大鼠和兔肾中 H-K-ATPase β 亚基 mRNA 的原位杂交。
DOI：
10.1152/ajprenal.1995.269.3.f345
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
McGuigan,JE
中科院分区：
文献类型：
--
作者：
Campbell-Thompson,ML;Verlander,JW;Curran,KA;Campbell,WG;Cain,BD;Wingo,CS;McGuigan,JE
一些研究人员通过各种技术证明了肾脏集合管中存在H-K-腺苷三磷酸酶(H-K-ATPase)，表明该酶在调节肾脏的酸碱平衡和钾排泄方面发挥着重要的生理作用。本研究用非放射性原位杂交技术对H-K-ATPaseβ亚基mRNA在大鼠和兔肾中的表达细胞进行了定位。以兔肾H-K-ATPaseβ亚基570bpDNA片段为模板，体外转录制备地高辛标记的核探针。Northern印迹杂交显示大鼠胃含氧酸粘膜和肾脏中均有转录。肾组织切片原位杂交结果显示，H-K-ATPaseβ亚基在大鼠和兔肾上皮细胞中均有分布，包括连接段、皮质和髓质集合管的间质细胞、外髓集合管内条的主细胞和内髓集合管细胞。这些观察结果提供了H-K-ATPaseβ亚单位mRNA存在于肾脏集合管的证据。这一信息的分布与H-K-ATPase在外髓和内髓集合管吸收碳酸氢盐中的作用是一致的。
Through a variety of techniques, several investigators have demonstrated the presence of an H-K-adenosinetriphosphatase (H-K-ATPase) enzyme in the renal collecting duct, suggesting that this enzyme serves an important physiological role in the regulation of acid-base balance and potassium excretion by the kidney. The present study was designed to localize cells expressing H-K-ATPase beta-subunit mRNA in rat and rabbit kidney by nonradioactive in situ hybridization. A 570-bp DNA fragment of rabbit renal H-K-ATPase beta-subunit was used to produce digoxigenin-labeled riboprobes by in vitro transcription. Northern blot hybridization demonstrated transcripts in rat gastric oxyntic mucosa and kidney. In situ hybridization on kidney tissue sections demonstrated H-K-ATPase beta-subunit mRNA localization in epithelial cells, including intercalated cells in the connecting segment and cortical and medullary collecting duct, principal cells in the inner stripe of the outer medullary collecting duct, and inner medullary collecting duct cells in both the rat and the rabbit. These observations provide evidence that H-K-ATPase beta-subunit mRNA is present throughout the collecting duct of the kidney. The distribution of this message is consistent with a role for H-K-ATPase in bicarbonate absorption in both the outer and inner medullary collecting duct.