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通过原位杂交技术检测血小板生成素mRNA在肝脏中的细胞定位。

Cellular localization of thrombopoietin mRNA in the liver by in situ hybridization.

作者信息

Nomura S, Ogami K, Kawamura K, Tsukamoto I, Kudo Y, Kanakura Y, Kitamura Y, Miyazaki H, Kato T

机构信息

Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan.

出版信息

Exp Hematol. 1997 Jul;25(7):565-72.

PMID:9216731
Abstract

The expression of thrombopoietin (TPO) mRNA is observed in several tissues, including liver, kidney, brain, skeletal muscle, intestine, spleen, and bone marrow. Among these organs, the highest expression of TPO mRNA is detected in the liver. We identified cells producing TPO by means of in situ hybridization of adult rat liver using digoxigenin-11-UTP-labeled cRNA probes. We found that the cells expressing TPO mRNA also expressed serum albumin mRNA. TPO mRNA was detected in parenchymal cells (hepatocytes) but not in non-parenchymal cells (including endothelial cells, epithelial cells, and so forth). To determine the location of TPO expression in embryogenesis, sections of fetal mice were further analyzed by in situ hybridization. TPO mRNA was detected only in hepatocytes of fetal liver, which was also the major site of hematopoiesis. The expression of TPO mRNA in fetal liver was observed from 12.5 days postcoitus. Northern blot analysis showed that mouse liver transcribed the same size of TPO mRNA in the fetus and in the adult. These results clearly demonstrate that hepatocytes are the primary site of TPO production in the liver from fetus to adult.

摘要

在包括肝脏、肾脏、大脑、骨骼肌、肠道、脾脏和骨髓在内的多种组织中均观察到血小板生成素(TPO)mRNA的表达。在这些器官中,TPO mRNA的表达在肝脏中最高。我们使用地高辛素-11-UTP标记的cRNA探针通过成年大鼠肝脏的原位杂交来鉴定产生TPO的细胞。我们发现表达TPO mRNA的细胞也表达血清白蛋白mRNA。在实质细胞(肝细胞)中检测到TPO mRNA,而在非实质细胞(包括内皮细胞、上皮细胞等)中未检测到。为了确定胚胎发育过程中TPO表达的位置,通过原位杂交进一步分析了胎鼠切片。仅在胎肝的肝细胞中检测到TPO mRNA,而胎肝也是造血的主要部位。在交配后12.5天观察到胎肝中TPO mRNA的表达。Northern印迹分析表明,小鼠肝脏在胎儿和成年期转录的TPO mRNA大小相同。这些结果清楚地表明,从胎儿到成年,肝细胞是肝脏中TPO产生的主要部位。

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