Zusammenfassung
Vorinkubation (30 min) von Rattendünndarmschnitten in Na+-freiem Puffer zerstört ihre Fähigkeit, L-Phenylalanin gegen einen Konzentrationsgradienten zu transportieren. Werden die Na+-Ionen durch Kalium, Lithium oder Tris-Ionen ersetzt, tritt ein völliger Verlust der Transportfähigkeit ein; werden sie dagegen durch Cholin-Ionen ersetzt, tritt eine weniger starke Schädigung ein. Vorinkubation in einem 1:1-Gemisch von Krebs-Puffer und K+-ersetztem Puffer bewirkt ebenfalls eine Verminderung der Transportfähigkeit.
Diese Ergebnisse können einem Mangel an verstoffwechselbarem Substrat innerhalb der Zelle während der Abwesenheit von Natrium nicht zugeschrieben werden. Sie lassen sich am besten durch die Annahme erklären, daß während der Vorinkubation die Natrium-Pumpe, die eng mit der Aminosäureaufnahme gekoppelt ist, zerstört wird. Wenn das Gewebe nach dieser Vorinkubation in Na+-haltige Lösung gebracht wird, dringen diese Ionen in die Zellen ein und zerstören die charakteristischen Eigenschaften der Zellmembran. Die Permeabilität des Gewebes für markiertes Natrium ist nach einer Vorinkubation in K+-reichem Medium signifikant größer gegenüber Kontroll-Versuchen. Entsprechend verhält sich die Permeabilität von Mannit.
Der Verlust der Transportfähigkeit nach Vorinkubation in Na+-freier Lösung scheint artspezifisch zu sein; Im Gegensatz zu Schnittpräparaten der Ratte und der Maus zeigen solche des Kaninchendünndarms diese Erscheinungen nicht. Nach Angaben der Literatur findet sich ein solcher Verlust der Transportfähigkeit ebenfalls nicht bei Dünndarmpräparaten der Kröte und des Goldhamsters. Da alle diese Gewebe ein Strophanthin-empfindliches Natrium-Transportsystem besitzen, wird diese enge Kopplung zwischen Empfindlichkeit gegenüber Na+-freier Vorinkubation und Unempfindlichkeit gegenüber Strophanthin als weiterer Hinweis für unterschiedliche Natrium-Transportsysteme bei verschiedenen Tierarten gewertet.
Summary
Preincubation of excised rings of rat intestine in sodium-free buffer for 30 min destroys the capacity of the tissue to absorb L-phenylalanine against a concentration gradient during a subsequent incubation in the presence of sodium ions. Use of potassium, lithium, or Tris ions as substituents for sodium causes complete loss of transport capacity, whereas the use of choline ions is somewhat less damaging. Preincubation in a 1:1 mixture of Krebs buffer and Na+-free, K+-substituted buffer also induces a reduction in subsequent transport capacity.
These results cannot be assigned to lack of metabolisable substrate within the cells when sodium is absent from the preincubation medium. They are best explained by suggesting that the sodium pump which is intimately linked with amino-acid accumulation within the epithelial cells is destroyed during the preincubation. Then when the tissue is replaced in a Na+-containing solution, the cells are flooded with these ions, which may then be responsible for the destruction of the characteristic properties of the cell membrane. Permeation of the tissue with radioactive sodium after a preincubation in K+-rich medium is significantly greater than in control samples, as is the penetration of mannitol.
This loss of activity following preincubation in the absence of sodium ions is partly species-specific; it is present in the rat and the mouse, and absent from the rabbit intestine (and according to the literature, from the toad and the hamster). Since the species from which the effect is absent have intestines possessing ouabainsensitive sodium pumps that are involved in non-electrolyte transport, it is suggested that this correlation between sensitivity to Na+-free preincubation and lack of susceptibility to ouabain provides further evidence for qualitative differences between the sodium-pump mechanisms in different animal species.
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This work was supported by a grant from the Deutsche Forschungsgemeinschaft, during the tenure of a visiting fellowship.
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Robinson, J.W.L. The loss of intestinal transport capacity following preincubation in sodium-free media in vitro. Pflügers Archiv 294, 182–200 (1967). https://doi.org/10.1007/BF00363605
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DOI: https://doi.org/10.1007/BF00363605