Perret C, Desplan C, Thomasset M
Eur J Biochem. 1985 Jul 1;150(1):211-7. doi: 10.1111/j.1432-1033.1985.tb09009.x.
In view of the possible physiological importance of the 9-kDa cholecalcin (a 9000-Mr cholecalciferol-induced calcium-binding protein) in the intestinal transport of calcium in mammals, the gene expression of this protein has been analysed. Its regulation in the digestive tract of the growing rat by calcitriol (1,25-dihydroxycholecalciferol) was studied using a specific cloned [32P]cDNA to 9-kDa cholecalcin. Northern hybridisation studies show that the cDNA sequence hybridises to a single 500-600-nucleotide species throughout the digestive tract and therefore demonstrate identical 9-kDa-cholecalcin mRNA processing in the whole of the intestine and caecum. The highest concentrations of cholecalcin mRNA occur in the duodenum, proximal jejunum and caecum. The observed differences in 9-kDa-cholecalcin mRNA levels correlate well with both the in vivo variations in cholecalcin itself and with the known intestinal sites of calcium absorption. The whole intestine is able to respond to exogenous calcitriol but the response of the distal intestine and caecum, as measured by the increase in cholecalcin mRNA and corresponding protein, was proportionally higher than in the duodenum. The rapid production of fully functional cholecalcin mRNA, which was detectable as early as 1 h after a single dose of calcitriol to vitamin-D-deficient rats, provides convincing evidence that calcitriol increases 9-kDa cholecalcin production by increasing cholecalcin gene expression at the transcriptional level.
鉴于9 kDa胆钙化蛋白(一种分子量为9000的胆钙化醇诱导的钙结合蛋白)在哺乳动物肠道钙转运中可能具有的生理重要性,对该蛋白的基因表达进行了分析。使用针对9 kDa胆钙化蛋白的特异性克隆[32P]cDNA,研究了生长中大鼠消化道中该蛋白受骨化三醇(1,25 - 二羟基胆钙化醇)的调节情况。Northern杂交研究表明,该cDNA序列在整个消化道中与单一的500 - 600核苷酸种类杂交,因此证明在整个肠道和盲肠中9 kDa胆钙化蛋白mRNA的加工过程相同。胆钙化蛋白mRNA浓度最高的部位是十二指肠、空肠近端和盲肠。观察到的9 kDa胆钙化蛋白mRNA水平差异与胆钙化蛋白本身的体内变化以及已知的钙吸收肠道部位密切相关。整个肠道都能够对外源性骨化三醇作出反应,但以胆钙化蛋白mRNA和相应蛋白的增加来衡量,远端肠道和盲肠的反应比十二指肠的反应比例更高。早在给维生素D缺乏的大鼠单次注射骨化三醇后1小时就能检测到完全功能性胆钙化蛋白mRNA的快速产生,这提供了令人信服的证据,表明骨化三醇通过在转录水平增加胆钙化蛋白基因表达来提高9 kDa胆钙化蛋白的产生。