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Studies in kidney tubulogenesis

Cytochemical localization of phosphatase and dehydrogenase activities during the formation of tubules in vitro

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Summary

The formation of secretory tubules in the embryonic mouse metanephrogenic mesenchyme in vitro has been produced with embryonic spinal cord serving as inductor. The activity of NADH-tetrazolium reductase, acid phosphatase, thiamine pyrophosphatase and ATP-ase activities was assayed cytochemically on frozen sections of cold-formalin fixed explants.

What was apparently the first phase of differentiation of kidney tubules became evident in the form of cell condensations. These were accompanied by an increased NADH-tetrazolium reductase activity, suggesting the maturation of both mitochondria (particulate reaction product) and endoplasmic reticulum (homogeneous staining). Furthermore, in the young explants there were visible areas of distinct enzyme activity, with no histological characteristics of the cell condensations. It was considered that these probably represented the earliest phase of differentiation detected by microscopic technique. Thiamine pyrophosphatase activity appeared in the Golgi apparatus of the cells during their organization into tubular epithelium and the formation of lumen. A subsequent increase in acid phosphatase activity of the cytoplasm was noticed. All of the active cytoplasmic organelles, marked by the enzyme activity cytochemically demonstrable, were moved to the apical side of the tubule cells in the course of their cytodifferentiation. After seven days of cultivation, the ATP-ase activity appeared first both in the cell membranes and in the cytoplasm of the tubule cells. At this phase of development, the kidney tubules induced in vitro thus displayed many of the cytochemical characteristics known to exist in the secretory tubules of the mature kidney.

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Supported by grant from the Sigrid Jusélius Foundation, by grant from Finnish Medical Research Council and by grant C-5347 from the National Cancer Institute, National Institutes of Health, Public Health Service to Prof. Toivonen and Dr. Saxén.

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Rapola, J., Niemi, M. Studies in kidney tubulogenesis. Z. Anat. Entwickl. Gesch. 124, 309–320 (1965). https://doi.org/10.1007/BF00523515

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