Clinical Investigation

Kidney International (1973) 3, 315–326; doi:10.1038/ki.1973.50

Transport of peroxidase in flounder kidney tubules studied by electron microscope histochemistry

Peter D Ottosen1 and Arvid B Maunsbach1

1Department of Cell Biology, Institute of Anatomy, University of Aarhus, Aarhus, Denmark

Correspondence: Dr Peter D Ottosen, Department of Cell Biology, Institute of Anatomy, University of Aarhus, DK-8000 Aarhus, Denmark.

Received 14 November 1972; Revised 3 January 1973.

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Abstract

Transport of peroxidase in flounder kidney tubules studied by electron microscope histochemistry. The aim of the present study was to determine at the ultrastructural level how protein is transported through the renal tubule wall. Flounders were injected intravenously with peroxidase, and renal tubules dissected and incubated in vitro. Peroxidase was located by electron microscope histochemistry and determined biochemically. At the start of incubation peroxidase was located 1) in the extracellular space in the tubule wall (intercellular space and basal invaginations) and 2) in apical cytoplasmic vacuoles. During the first hour of incubation peroxidase located in the extracellular space disappeared and simultaneously the peroxidase content in the incubation fluid increased. Peroxidase located in the apical vacuoles was transferred to lysosomes. In control experiments tubules from uninjected fish were incubated in a medium containing peroxidase. Within 30 min peroxidase penetrated retrograde into the extracellular space across the basement membrane. The present study shows that peroxidase 1) can penetrate into the extracellular space via the basement membrane and 2) diffuse out into the peritubular fluid during incubation in vitro. The latter process may simulate transtubular transport of intact protein.

Transport de la péroxydase par le tube rénal du flet étudié par histochimie en microscopie électronique. Le but de ce travail est de déterminer, au niveau ultrastructur, la façon dont les protéines sont transportées à travers la paroi tubulaire rénale. Les flets ont reçu de la péroxydase par voie intraveineuse et les tubes rénaux ont été disséqués et incubés in vitro. La péroxydase a été localisée par des méthodes histochimiques en microscopie électronique et sa concentration déterminée biochimiquement. Au début de l'incubation la péroxydase était localisée 1) dans l'espace extracellulaire de la paroi tubulaire (espace intercellulaire et invaginations de la basale) et 2) dans les vacuoles apicales du cytoplasme. Pendant la première heure d'incubation la péroxydase localisée dans l'espace extracellulaire disparaît et, simultanément, le contenu en péroxydase du liquide d'incubation augmente. La péroxydase des vacuoles apicales est transférée aux lysosomes. Au cours des expériences témoins des tubules de poissons non injectés ont été incubés dans un milieu contenant de la péroxydase. En 30 minutes la péroxyadse a pénétré par voie rétrograde dans l'espace extracellulaire, à travers la membrane basale. Ce travail montre que la péroxydase 1) peut pénétrer dans l'espace extracellulaire à travers la membrane basale et 2) diffuse dans le liquide péritubulaire au cours de l'incubation in vitro. Ce dernier processus peut simuler un transport transtubulaire de protéine intacte.

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