Theofan G, King M W, Hall A K, Norman A W
Division of Biomedical Sciences, University of California, Riverside 92521.
Mol Cell Endocrinol. 1987 Dec;54(2-3):135-40. doi: 10.1016/0303-7207(87)90150-x.
The availability of specific cDNA probes to the chick intestinal calbindin-D28K (CaBP) mRNA has allowed us to assess the regulation of this mRNA in response to 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) administration. It has previously been demonstrated that dietary calcium and phosphorus can effect alterations in the steady-state intestinal levels of chick CaBP. We have examined whether or not perturbations in dietary calcium and phosphorus have an effect on the expression of the intestinal mRNA coding for CaBP in the vitamin D-replete chick. We found altered protein levels of CaBP as expected; however there was surprisingly no difference in steady-state CaBP mRNA levels between the different dietary groups. These data suggest that calcium and phosphorus regulation of CaBP occurs at a post-transcriptional level. In addition, we have examined what effect dietary manipulations of calcium and phosphorus levels have on the response of the vitamin D-replete intestine to 1,25(OH)2D3 administration as assessed by CaBP mRNA changes. Administration of 1,25(OH)2D3 to vitamin D-replete chicks maintained on normal calcium and phosphorus levels resulted in a less than 2-fold increase in CaBP mRNA levels. Previous studies have demonstrated that receptor occupancy goes up 6-fold under these conditions; therefore there is apparently a very tight regulation of CaBP gene activity. 1,25(OH)2D3 administration to chicks raised on either low calcium, high calcium, or low phosphorus vitamin D-replete diets similarly showed only small changes in the intestinal CaBP mRNA levels; however there seemed to be qualitative differences in response attributable to the dietary alterations.
有了针对鸡肠道钙结合蛋白-D28K(CaBP)mRNA的特异性cDNA探针,使我们能够评估该mRNA对1,25-二羟维生素D3(1,25(OH)2D3)给药的反应调节情况。先前已经证明,日粮中的钙和磷可影响鸡CaBP的肠道稳态水平变化。我们研究了日粮中钙和磷的扰动是否会对维生素D充足的鸡中编码CaBP的肠道mRNA的表达产生影响。我们发现CaBP的蛋白水平如预期那样发生了变化;然而,不同日粮组之间的CaBP mRNA稳态水平出人意料地没有差异。这些数据表明,CaBP的钙和磷调节发生在转录后水平。此外,我们还研究了日粮中钙和磷水平的改变对维生素D充足的肠道对1,25(OH)2D3给药反应的影响(通过CaBP mRNA变化来评估)。给维持在正常钙和磷水平的维生素D充足的小鸡注射1,25(OH)2D3,导致CaBP mRNA水平增加不到2倍。先前的研究表明,在这些条件下受体占有率会增加6倍;因此,CaBP基因活性显然受到非常严格的调节。给饲养在低钙、高钙或低磷维生素D充足日粮上的小鸡注射1,25(OH)2D3,同样显示肠道CaBP mRNA水平只有很小的变化;然而,由于日粮改变,反应似乎存在质的差异。