Couvineau A, Darmoul D, Blais A, Rouyer-Fessard C, Daviaud D, Voisin T, Paris H, Rouot B, Laburthe M
Institut National de la Santé et de la Recherche Médicale (INSERM) U. 239, Faculté de Médecine X. Bichat, Paris, France.
Am J Physiol. 1992 Jun;262(6 Pt 1):C1478-84. doi: 10.1152/ajpcell.1992.262.6.C1478.
The expression of subunits of the guanine nucleotide regulatory protein that mediates hormonal stimulation of adenylyl cyclase (Gs) and of the guanine nucleotide regulatory protein that mediates hormonal inhibition of adenylyl cyclase (Gi) was studied during cell migration and differentiation in the rat small intestine crypt-villus axis. Proliferative crypt cells were separated from nonproliferative mature villus cells and the following data were obtained: 1) alpha s subunits were more abundant in crypt cells than in villus cells as evidenced by cholera toxin-catalyzed [32P]NAD ribosylation and Western blotting of this relative molecular weight (M(r)) 42,000 protein; 2) alpha i2- and alpha i3-subunits (M(r) 40,000 and M(r) 41,000, respectively) were preferentially expressed in villus cells as evidenced by pertussis toxin-catalyzed [32P]NAD ribosylation and Western blotting (alpha i1-subunit was not detectable in intestinal epithelium by using these techniques); 3) Western blotting showed a higher expression of the common beta- (M(r) 36,000) subunit of G proteins in villus cells than in crypt cells; and 4) Northern blot analysis using an alpha s-subunit oligonucleotide probe showed a 1.9-kb mRNA that was more abundant in crypt cells than in villus cells. In contrast, alpha i2- and alpha i3-mRNA species (2.3 and 3.5 kb, respectively), analyzed by using specific cDNA probes, were much more abundant in villus cells than in crypt cells. Finally, two beta-subunit mRNA species of 3.3 and 1.8 kb were detectable in intestinal epithelial cells and were more abundant in villus cells than in crypt cells.(ABSTRACT TRUNCATED AT 250 WORDS)
在大鼠小肠隐窝 - 绒毛轴的细胞迁移和分化过程中，研究了介导激素刺激腺苷酸环化酶的鸟嘌呤核苷酸调节蛋白（Gs）亚基以及介导激素抑制腺苷酸环化酶的鸟嘌呤核苷酸调节蛋白（Gi）亚基的表达。将增殖的隐窝细胞与非增殖的成熟绒毛细胞分离，并获得以下数据：1）通过霍乱毒素催化的[32P]NAD核糖基化以及对这种相对分子质量（M(r)）为42,000的蛋白质进行蛋白质印迹分析表明，αs亚基在隐窝细胞中比在绒毛细胞中更丰富；2）通过百日咳毒素催化的[32P]NAD核糖基化以及蛋白质印迹分析表明，αi2 - 和αi3 - 亚基（分别为M(r) 40,000和M(r) 41,000）优先在绒毛细胞中表达（使用这些技术在肠上皮中未检测到αi1 - 亚基）；3）蛋白质印迹分析显示，G蛋白的共同β - 亚基（M(r) 36,000）在绒毛细胞中的表达高于隐窝细胞；4）使用αs - 亚基寡核苷酸探针进行的Northern印迹分析显示，一种1.9 - kb的mRNA在隐窝细胞中比在绒毛细胞中更丰富。相比之下，使用特异性cDNA探针分析的αi2 - 和αi3 - mRNA种类（分别为2.3和3.5 kb）在绒毛细胞中比在隐窝细胞中丰富得多。最后，在肠上皮细胞中可检测到两种3.3和1.8 kb的β - 亚基mRNA种类，并且在绒毛细胞中比在隐窝细胞中更丰富。（摘要截短于250字）
