Localization of cytochrome P-450 4A isoforms along the rat nephron
Localization of cytochrome P-450 4A isoforms along the rat nephron
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DOI：
10.1152/ajprenal.1998.274.2.f395
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发表时间：
1998-02-01
影响因子：
4.2
通讯作者：
Roman, RJ
中科院分区：
文献类型：
--
作者：
Ito, O;Alonso-Galicia, M;Roman, RJ
使用逆转录和聚合酶链反应在从 Sprague-Dawley 大鼠肾脏显微解剖的各个肾单位节段和肾小球前小动脉中检查了负责形成 20-羟基二十碳四烯酸 (80-HETE) 的 P-450 4A 亚型的表达。从全肾提取的RNA中可检测到细胞色素P-450 4A1、4A2、4A3和4A8 mRNA的表达。雄性和雌性大鼠肾脏中 P-450 4A1、4A3 和 4A8 mRNA 的表达相似，而雄性大鼠肾脏中 4A2 mRNA 的表达量是雌性大鼠肾脏的四倍。在单肾单位水平，在肾小球前小动脉或任何肾单位节段中均未检测到 P-450 4A1 mRNA。 P-450 4A2 mRNA 在肾小球前小动脉、肾小球、近曲小管 (PCT)、近端直管 (PST)、髓质厚升肢 (MTAL)、皮质厚升肢 (CTAL)、皮质集合管 (CCD)、外髓集合管 (OMCD) 和内髓集合管 (OMCD) 中很容易检测到。 髓质集合管（IMCD）。 P-450 4A3 mRNA 在每个肾单位节段中均检测到，但在肾小球前小动脉中几乎检测不到该亚型的表达。在肾小球、PCT、PST、CTAL和CCD中检测P-450 4A8 mRNA的表达。在肾小球前小动脉、MTAL、OMCD 或 IMCD 中未检测到它。使用 P-450 4A 抗体进行的免疫印迹分析显示近曲小管中 P-450 4A 蛋白的强信号。在肾小球、MTAL 和肾小球前小动脉中也观察到较小的信号，但在 IMCD 中未检测到信号。在用该抗体免疫染色的肾脏切片中观察到类似的 P-450 4A 蛋白表达模式。这些结果表明，P-450 4A 亚型在大鼠肾脏中的表达具有性别依赖性，并且不同的 P-450 4A 亚型在大鼠的各个肾单位节段和肾脉管系统中表达。
The expression of P-450 4A isoforms responsible for the formation of 20-hydroxyeicosatetraenoic acid (80-HETE) was examined using the reverse transcription and polymerase chain reaction in various nephron segments and preglomerular arterioles microdissected from the kidneys of Sprague-Dawley rats. Expression of cytochrome P-450 4A1, 4A2, 4A3, and 4A8 mRNA could be detected in RNA extracted from the whole kidney. The expression of P-450 4A1, 4A3, and 4A8 mRNA was similar in the kidney of male and female rats, whereas the expression of 4A2 mRNA was fourfold greater in the kidney of male vs. female rats. At the single-nephron level, P-450 4A1 mRNA could not be detected in either preglomerular arterioles or any nephron segments. P-450 4A2 mRNA was readily detected in preglomerular arterioles, glomeruli, proximal convoluted tubule (PCT), proximal straight tubule (PST), medullary thick ascending limb (MTAL), cortical thick ascending limb (CTAL), cortical collecting duct (CCD), outer medullary collecting duct (OMCD), and inner medullary collecting duct (IMCD). P-450 4A3 mRNA was also detected in every nephron segment, but the expression of this isoform was barely detectable in preglomerular arterioles. The expression of P-450 4A8 mRNA was detected in the glomerulus, PCT, PST, CTAL, and CCD. It was not detectable in preglomerular arterioles, MTAL, OMCD, or IMCD. Immunoblot analysis using a P-450 4A antibody exhibited a strong signal for P-450 4A protein in the proximal tubule. Smaller signals were also observed in glomerulus, MTAL, and preglomerular arterioles, but no signal could be detected in the IMCD. A similar pattern of P-450 4A protein expression was seen in kidney sections immunostained with this antibody. These results indicate that the expression of P-450 4A isoforms in the kidney of rats is sex dependent and that different P-450 4A isoforms are expressed throughout various nephron segments and the renal vasculature of rats.