The establishment of a primary culture system of proximal tubule segments using specific markers from normal mouse kidneys.
The establishment of a primary culture system of proximal tubule segments using specific markers from normal mouse kidneys.
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DOI：
10.3390/ijms13045098
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发表时间：
2012
影响因子：
5.6
通讯作者：
Kobori H
中科院分区：
文献类型：
--
作者：
Kamiyama M;Garner MK;Farragut KM;Kobori H
近端小管是肾脏中血管紧张素原基因和蛋白表达最高的区域，在肾素-血管紧张素系统中起着至关重要的作用。为了更详细地研究血管紧张素原在肾脏中的表达调节，需要将近端小管与肾单位的其余部分准确地分离出来，并将其分为三个部分。这项研究的目的是设计一种新的方法，使用特定的标记将近端小管细胞分离到三个近端小管段，并检测每一段血管紧张素原的表达。取C57BL/6J小鼠肾脏。从适当密度的Percoll梯度溶液区域抽吸近端小管。然后使用片段特异性膜蛋白将近端小管分成三个片段，然后每个片段用不同的特异性标记(片段1的钠-葡萄糖转运体2；片段2的碳酸氢酶IV；片段3的胞外三磷酸腺苷酶)来表征。成功地将近端肾小管分离成3段，证实了2、3段血管紧张素原mRNA和3段血管紧张素原蛋白的存在。该方案将有助于进一步研究肾内肾素-血管紧张素系统的详细机制。
The proximal tubule contains the highest expression of angiotensinogen mRNA and protein within the kidney and plays a vital role in the renal renin-angiotensin system. To study the regulation of angiotensinogen expression in the kidney in more detail, the proximal tubule needs to be accurately isolated from the rest of the nephron and separated into its three segments. The purpose of this study was to design a novel protocol using specific markers for the separation of proximal tubule cells into the three proximal tubule segments and to determine angiotensinogen expression in each segment. Kidneys were removed from C57BL/6J mice. The proximal tubules were aspirated from region of a Percoll gradient solution of the appropriate density. The proximal tubule was then separated into its three segments using segment-specific membrane proteins, after which each segment was characterized by a different specific marker (sodium-glucose transporter 2 for Segment 1; carbonic anhydrase IV for Segment 2; ecto-adenosine triphosphatase for Segment 3). The isolation of proximal tubules into three segments was successful, and angiotensinogen mRNA in Segment 2 and 3 and angiotensinogen protein in all three segments were confirmed. This protocol will be helpful for future studies of the detailed mechanisms of the intrarenal renin-angiotensin system.
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影响因子：
7.8
作者：
CHUMAN, L;FINE, LG;SAIER, MH
通讯作者：
SAIER, MH
影响因子：
19.6
作者：
DARBY, IA;CONGIU, M;COGHLAN, JP
通讯作者：
COGHLAN, JP
影响因子：
5.5
作者：
Christensen, BM;Marples, D;Nielsen, S
通讯作者：
Nielsen, S
影响因子：
8.3
作者：
Prieto-Carrasquero, MC;Harrison-Bernard, LM;Navar, LG
通讯作者：
Navar, LG
DOI：
10.1007/s00259-005-1793-0
发表时间：
2005-10-01
影响因子：
9.1
作者：
Melis, M;Krenning, EP;de Jong, M
通讯作者：
de Jong, M