Localization of erythropoietin mRNA in the rat kidney by polymerase chain reaction
Localization of erythropoietin mRNA in the rat kidney by polymerase chain reaction
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聚合酶链式反应对大鼠肾脏促红细胞生成素 mRNA 的定位
DOI：
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发表时间：
1994
影响因子：
4
通讯作者：
N. Abraham
中科院分区：
文献类型：
--
作者：
J. da Silva;M. Schwartzman;Alvin Goodman;R. Levere;N. Abraham
促红细胞生成素（Epo）是肾细胞分泌的一种糖蛋白，在红细胞生成过程中起着重要的调节作用。通过免疫组织化学研究和原位杂交定位Epo生产尚未明确建立，仍然是一个有争议的问题。采用逆转录酶/聚合酶链式反应（RT/PCR）偶联方法测定对照和CoCl2处理大鼠总RNA中Epo和甘油醛3 -脱氢酶（GAPDH） mRNA水平。根据扩增产物的数量，在CoCl2处理的大鼠肾脏中，Epo mRNA水平高出数倍。相比之下，对照组和CoCl2处理大鼠的GAPDH mRNA水平相似。RT/PCR法检测肾细胞切片中Epo和GAPDH mRNA表达水平。在对照组或CoCl2处理的大鼠中，所有测试的肾单元段都缺乏可检测到的Epo mRNA水平。另一方面，小管周围细胞（毛细血管部分：传入/传出小动脉，直血管）是唯一检测到Epo mRNA的细胞。使用GAPDH特异性引物，RT/PCR方法可以在所有未表达Epo mRNA的微解剖肾细胞片段中鉴定GAPDH mRNA。因此，显微解剖肾单元片段和RT/PCR相结合使我们能够检测到所有肾单元片段中的GAPDH mRNA群体，而未能检测到除毛细血管部分外所有片段中的Epo mRNA，这是由于Epo基因在该部分的特异性和局部表达。
Erythropoietin (Epo) is a glycoprotein secreted by kidney cells which plays an important role in the regulation of erythropoiesis. Localization of the Epo production by immunohistochemical studies and in situ hybridization has not been definitively established and is still a matter of controversy. Epo and glyceraldehyde 3‐dehydrogenase (GAPDH) mRNA levels were determined in total RNA isolated from control and CoCl2‐treated rats using a coupled reverse transcriptase/polymerase chain reaction method (RT/PCR). As indicated by the amount of amplification product, Epo mRNA levels were several‐fold higher in CoCl2‐treated rat kidney. In contrast, GAPDH mRNA levels were similar in control and CoCl2‐treated rats. This RT/PCR method was also used to assess the level of Epo and GAPDH mRNA in microdissected nephron segments. All nephron segments tested lacked any detectable levels of Epo mRNA in either control or CoCl2‐treated rats. On the other hand, peritubular cells (capillary fraction: afferent/efferent arteriole, vasa recta) were the only cells where the Epo mRNA was detected. Using a specific primer for GAPDH, the RT/PCR method could identify GAPDH mRNA in all microdissected nephron segments where the Epo mRNA was not expressed. Thus, a combination of microdissected nephron segments and RT/PCR enabled us to detect GAPDH mRNA populations in all nephron segments, whereas the failure to detect Epo mRNA in all segments but the capillary fraction, is due to the specific and localized expression of the Epo gene to this fraction.
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影响因子：
4.3
作者：
John B. Sharefkin;Scott L. Diamond;S. Eskin;L. McIntire;Carl W. Dieffenbach
通讯作者：
John B. Sharefkin;Scott L. Diamond;S. Eskin;L. McIntire;Carl W. Dieffenbach
肾促红细胞生成素信使 RNA 的生理调节和组织定位。
影响因子：
20.3
作者：
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通讯作者：
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影响因子：
2.6
作者：
Lin,JH;Grandchamp,B;Abraham,NG
通讯作者：
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影响因子：
20.3
作者：
S. Schuster;EV Badiavas;P. Costa-Giomi;R. Weinmann;A. Erslev;J. Caro
通讯作者：
S. Schuster;EV Badiavas;P. Costa-Giomi;R. Weinmann;A. Erslev;J. Caro
血红素诱导的人红白血病细胞分化过程中早期原癌基因的表达。
影响因子：
2.7
作者：
Griffin,MO;Nelson,JC;Abraham,NG
通讯作者：
Abraham,NG