Endogenous plasma membrane proteins are sorted from two sites in the human intestinal epithelial cell line Caco‐2. Apical proteins are transported from the Golgi apparatus to the apical domain along a direct pathway and an indirect pathway via the basolateral membrane. In contrast, basolateral proteins never appear in the apical plasma membrane. Here we report on the effect of the microtubule‐active drug nocodazole on the post‐synthetic transport and sorting of plasma membrane proteins. Pulse‐chase radiolabeling was combined with domain‐specific cell surface assays to monitor the appearance of three apical and one basolateral protein in plasma membrane domains. Nocodazole was found to drastically retard both the direct transport of apical proteins from the Golgi apparatus and the indirect transport (transcytosis) from the basolateral membrane to the apical cell surface. In contrast, neither the transport rates of the basolateral membrane nor the sorting itself were significantly affected by the nocodazole treatment. We conclude that an intact microtubular network facilitates, but is not necessarily required for, the transport of apical membrane proteins along the two post‐Golgi pathways to the brush border.
Abstract. Novel subcellular fractionation procedures and pulse-chase techniques were used to study the intracellular transport of the microvillar membrane hydrolases sucrase-isomaltase and dipeptidylpeptidase IV in the differentiated colon adenocarcinoma cell line Caco-2. The overall rate of transport to the cell surface was two fold faster for dipeptidylpeptidase IV than for sucrase-isomaltase, while no significant differences were observed in transport rates from the site of complex glycosylation to the brush border. The delayed arrival of sucrase-isomaltase in the compartment where complex glycosylation occurs was only in part due to exit from the endoplasmic reticulum. A major slow-down could be ascribed to maturation in and transit of this enzyme through the Golgi apparatus. These results suggest that the observed asynchronism is due to more than one rate-limiting step along the rough endoplasmic reticulum to trans-Golgi pathway.
Summary. In anesthetized, pontine-deeerebrated rabbits it was shown that the tendency of inspirations as well as exspirations to preferably begin within certain sections of the time course of a cardiac cycle, did still persist after bilateral vagotomy. This linking between cardiac and respiratory rhythm is compared with other linkings which have already been pointed out for rabbits. Reflections on eardiorespiratory synchronism in men are made.
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