REGULATION OF BRUSH-BORDER ENZYME-ACTIVITIES AND ENTEROCYTE MIGRATION RATES IN MOUSE SMALL-INTESTINE
REGULATION OF BRUSH-BORDER ENZYME-ACTIVITIES AND ENTEROCYTE MIGRATION RATES IN MOUSE SMALL-INTESTINE
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DOI：
10.1152/ajpgi.1992.262.6.g1047
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发表时间：
1992-06-01
影响因子：
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通讯作者：
DIAMOND, J
中科院分区：
文献类型：
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作者：
FERRARIS, RP;VILLENAS, SA;DIAMOND, J
我们采用了Weiser方法，以前用于大鼠和兔肠的肠上皮细胞，小鼠的小肠。通过组织学、形态学、酶促、组织化学和免疫细胞化学证据，该方法成功地去除小鼠肠上皮细胞顺序沿着隐窝绒毛轴，同时保持细胞活力和最小化细胞组分之间的混合。三种刷状缘酶[碱性磷酸酶（AP），蔗糖酶和γ-谷氨酰转肽酶（GGP）]的活性变化同时与饮食基质水平，肠道区域，和位置沿着隐窝-绒毛轴。所有这三种酶被证明是刺激的膳食底物：蔗糖酶由膳食蔗糖，AP和GGP由膳食蛋白质。我们还研究了细胞迁移率和寿命的放射自显影和我们的修改韦泽方法。通过这两种方法，注射[H-3]胸苷后短时间内几乎局限于隐窝细胞，而40-48小时后，它分布在整个绒毛，除了绒毛尖端的隐窝。绒毛高度降低两倍，从十二指肠到回肠，平行于区域细胞迁移率的下降，因为68小时的细胞寿命是独立的区域。当我们改变饮食中的碳水化合物和蛋白质水平，同时保持蛋白质高于维持水平时，细胞迁移率和细胞寿命都被证明与饮食无关。
We adapted the Weiser method, previously used to fractionate enterocytes of rat and rabbit intestine, to the much smaller intestine of mice. By histological, morphometric, enzymatic, histochemical, and immunocytochemical evidence, the method succeeded in removing mouse enterocytes sequentially along the crypt-villus axis while preserving cell viability and minimizing mixing among cell fractions. Activities of three brush-border enzymes [alkaline phosphatase (AP), sucrase, and gamma-glutamyl transpeptidase (GGP)] varied simultaneously with dietary substrate level, intestinal region, and position along the crypt-villus axis. All three enzymes proved to be stimulated by dietary substrate: sucrase by dietary sucrose, AP and GGP by dietary protein. We also studied cell migration rates and lifetimes by autoradiography and by our modified Weiser method. By both methods, injected [H-3]thymidine after short times was virtually confined to crypt cells, whereas after 40-48 h it was distributed from the crypt over the whole villus except for the villus tip. Villus height decreased twofold from duodenum to ileum, parallel to the regional decrease in cell migration rates because the cell lifetime of 68 h was independent of region. When we varied dietary carbohydrate and protein levels reciprocally while maintaining protein above the maintenance level, both cell migration rate and cell lifetime proved independent of diet.