Traber P G
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor.
Biochem Biophys Res Commun. 1990 Dec 31;173(3):765-73. doi: 10.1016/s0006-291x(05)80853-8.
The expression of sucrase-isomaltase mRNA was investigated along the crypt-villus axis of rat small intestine using differentially isolated cells and in situ hybridization. A partial rat sucrase-isomaltase cDNA was cloned which coded for a protein that was predicted to be 88% homologous to those encoded by the rabbit and human cDNAs. Southern blot analysis of rat genomic DNA indicated that the cDNA hybridized to a single gene. Northern blots of RNA extracted from subpopulations of intestinal epithelial cells that were isolated from villus and crypt compartments showed that this cDNA hybridized to a 6.5 kb band predominantly in villus RNA. In situ hybridization using 35[S]-labeled RNA probes demonstrated that autoradiographic grains were detected over eptithelial cells located on villi with the greatest number of grains located at the crypt-villus junction and in the lower to mid-villus region; from mid-villus to the villus tip there was a decline in sucrase-isomaltase mRNA. We conclude that expression of sucrase-isomaltase as enterocytes emerge from intestinal crypts is regulated primarily at the level of mRNA accumulation which, most likely, is a result of activation of sucrase-isomaltase gene transcription.
利用差异分离细胞和原位杂交技术,沿着大鼠小肠的隐窝-绒毛轴研究蔗糖酶-异麦芽糖酶mRNA的表达。克隆了大鼠蔗糖酶-异麦芽糖酶cDNA的一个片段,该片段编码的蛋白质预计与兔和人cDNA编码的蛋白质具有88%的同源性。大鼠基因组DNA的Southern印迹分析表明,该cDNA与单个基因杂交。从绒毛和隐窝区分离的肠上皮细胞亚群中提取的RNA进行Northern印迹分析,结果显示该cDNA主要与绒毛RNA中的一条6.5 kb条带杂交。使用35[S]标记的RNA探针进行原位杂交表明,在绒毛上的上皮细胞上检测到放射自显影颗粒,其中数量最多的颗粒位于隐窝-绒毛交界处和绒毛中下部区域;从绒毛中部到绒毛顶端,蔗糖酶-异麦芽糖酶mRNA水平下降。我们得出结论,随着肠上皮细胞从肠隐窝中出现,蔗糖酶-异麦芽糖酶的表达主要在mRNA积累水平上受到调节,这很可能是蔗糖酶-异麦芽糖酶基因转录激活的结果。