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兔肠道中质膜钙泵PMCA1转录本的细胞及区域表达

Cellular and regional expression of transcripts of the plasma membrane calcium pump PMCA1 in rabbit intestine.

作者信息

Freeman T C, Howard A, Bentsen B S, Legon S, Walters J R

机构信息

Sanger Centre, Hinxton, Cambridge, United Kingdom.

出版信息

Am J Physiol. 1995 Jul;269(1 Pt 1):G126-31. doi: 10.1152/ajpgi.1995.269.1.G126.

Abstract

The plasma membrane Ca(2+)-pumping adenosinetriphosphatase (PMCA) is the energy-dependent step in the active vitamin D-dependent absorption of dietary Ca2+ by the enterocyte. Studies of the various PMCA genes and splicing variants in humans and rats have indicated that the isoform known as PMCA1b is the predominant form expressed in small intestine. Using an oligonucleotide probe, we have studied the regional and cellular distribution of PMCA1 transcripts in rabbit intestinal tissues by in situ hybridization. On small intestinal RNA blots, this hybridized to species similar in size to those detected by PMCA1-specific cDNA probes; an additional larger transcript was present in rabbit than in rat or human. In situ hybridization signals were principally in the enterocyte population of the mucosa and were maximal in differentiating enterocytes on the lower part of the villus, a pattern similar to that previously demonstrated for other nutrient transporters. Reflecting the capacity of the different small intestinal segments to transport Ca2+, much higher levels of transcript were detected by both methods proximally (in duodenum) than distally (in jejunum and ileum) and were also higher in cecum and ascending colon mucosa than in descending colon. We conclude that as enterocytes differentiate in regions that absorb Ca2+, they express high levels of mRNA for PMCA1. These results confirm the importance of transcriptional regulation of this gene for active Ca2+ absorption.

摘要

质膜钙泵三磷酸腺苷酶(PMCA)是肠细胞对膳食钙进行维生素D依赖的主动吸收过程中的能量依赖步骤。对人和大鼠中各种PMCA基因及剪接变体的研究表明,被称为PMCA1b的同工型是小肠中表达的主要形式。我们使用寡核苷酸探针,通过原位杂交研究了兔肠道组织中PMCA1转录本的区域和细胞分布。在小肠RNA印迹上,该探针与大小与PMCA1特异性cDNA探针检测到的相似的条带杂交;与大鼠或人类相比,兔中还存在一个额外的较大转录本。原位杂交信号主要存在于黏膜的肠细胞群体中,在绒毛下部正在分化的肠细胞中信号最强,这一模式与先前其他营养转运蛋白的模式相似。反映不同小肠段运输钙的能力,两种方法在近端(十二指肠)检测到的转录本水平比远端(空肠和回肠)高得多,盲肠和升结肠黏膜中的转录本水平也比降结肠高。我们得出结论,随着肠细胞在吸收钙的区域分化,它们会表达高水平的PMCA1 mRNA。这些结果证实了该基因转录调控对钙的主动吸收的重要性。

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