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Untersuchungen zum Proteinstoffwechsel des Wildtyps und der Letalmutante (ltr) von Drosophila melanogaster

Studies on protein metabolism in the wildtype and the lethal mutant “ltr” of drosophila melanogaster

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Summary

  1. 1.

    Comparative studies on the protein metabolism in larvae of different developmental stages of the wild-type (+/+) and the lethal mutant “lethal-translucida” (ltr; 3, 20.7 ±0.8) of Drosophila melanogaster have been carried out. These included the changes of wet weight, hemolymph volume, content of hemolymph proteins, activity of protein synthesis of the fat body in vitro and protease activity of hemolymph and fat body.

  2. 2.

    The wet weight of wild-type larvae doubles from the third to the fourth day of age. During the same period there is a threefold increase in the lethals. The wet weight of the ltr-fat body is, however, only 33 % of that of the normals.

  3. 3.

    The concentration of hemolymph proteins in the wildtype larvae increases from 2.6% at 72 hrs of age to 6.9% at 96 hrs, whereas in the lethals the corresponding value is only 0.7%. Nevertheless, their total protein content of hemolymph per larva amounts to 78% of the wild-type value since the lethal individuals contain about eight times more hemolymph than the normals.

  4. 4.

    The in vitro experiments with fat body of young wildtype larvae showed a high incorporation rate of 14C-valine into proteins, which decreases rapidly as development proceeds.

  5. 5.

    The fat body of the ltr -homozygotes aged 90 to 130 hrs incorporates approximately the same amount of valine into proteins as the fat body of a 65hrs-old-wild-type larva. Even at the end of the larval period there is no decrease in the incorporation rate.

  6. 6.

    The fat body of both genotypes releases proteins synthesized in vitro into the incubation medium. Based on their electrophoretic mobility in starch and polyacrylamide gel some of them have been identified as hemolymph proteins.

  7. 7.

    The young wild-type larvae show a high incorporation rate of injected 14C-valine into proteins of the fat body, hemolymph and other tissues.

  8. 8.

    The old lethal larvae are capable of replacing within 24 hrs most of the hemolymph lost by puncture. This restituted hemolymph has again a protein content of 0.7%.

  9. 9.

    In lethals the protease activity of hemolymph is 7.7 times higher than that in wild-type larvae. The corresponding enzyme activity is four times higher for the fat body.

  10. 10.

    The present results demonstrated that the abnormal protein metabolism in the ltr-lethals is characterized by a high rate of protein synthesis as well as a rapid protein degradation. The possibility that this abnormality is causally related to some failure in the osmoregulation of the mutant larvae is suggested.

Zusammenfassung

  1. 1.

    Es wurden bei Larven des Wildtyps und der Letalmutante „letal-translucida“ (ltr; 3, 20,7 ±0,8) von Drosophila melanogaster in verschiedenen Entwicklungsstadien die Veränderungen folgender Größen untersucht: Frischgewicht, Hämolymphvolumen, Blutproteingehalt, Proteinsynthese-Aktivität des Fettkörpers in vitro und Proteasenaktivität von Blut und Fettkörper.

  2. 2.

    Das Frischgewicht der Normallarven verdoppelt sich im Zeitabschnitt 72–96 Std. Die Letallarven erreichen vor der Verpuppung ein 3 mal höheres Frischgewicht als die Normalen. Dagegen beträgt das Frischgewicht des ltr-Fettkörpers in diesem Stadium nur 33% des Normal-Gewichtes.

  3. 3.

    Der Gehalt an Blutproteinen steigt bei Normallarven von 2,6% (72 h) auf 6,9% (96 h). Bei verpuppungsreifen Letallarven enthält das Blut nur 0,7% Eiweiße. Pro Larve berechnet, ergibt dies 78% des totalen Blutproteingehaltes der Normalen, da die Letalen ein 8 mal größeres Blutvolumen haben.

  4. 4.

    In vitro Versuche mit Fettkörper junger Normallarven zeigen einen hohen Einbau von 14C-Valin in die Proteine, dann folgt bis zur Verpuppung eine starke Abnahme des Einbaues.

  5. 5.

    Der Fettkörper von ltr-Homozygoten (90–130 h) baut ungefähr gleichviel Valin in Proteine ein wie ein 65stündiger Normalfettkörper und zeigt keine Abnahme der Einbauleistung gegen das Ende der Larvalperiode.

  6. 6.

    Der Fettkörper beider Genotypen gibt in vitro gebildete Proteine ins Inkubationsmedium ab, die an Hand der elektrophoretischen Beweglichkeit zum Teil als Blutproteine identifiziert werden können.

  7. 7.

    Junge Normallarven zeigen einen höheren Einbau von injiziertem 14C-Valin in die Proteine des Fettkörpers, des Blutes und der übrigen Gewebe.

  8. 8.

    Alte Letallarven können das durch Punktieren verlorene Blut innerhalb 24 Std weitgehend ersetzen, der Proteingehalt der Ersatzhämolymphe beträgt wieder 0,7%.

  9. 9.

    Gegenüber Normalen ist bei den Letalen die Proteasenaktivität im Blut 7,7-, im Fettkörper-Homogenat 4 mal höher (pro mg Protein der Enzymprobe).

  10. 10.

    Nach den vorliegenden Resultaten besteht die Abnormität im Proteinstoffwechsel der ltr-Letalen darin, daß in großem Maße Proteine auf- und abgebaut werden. Es wird auf einen möglichen Zusammenhang zwischen dieser Störung und einem Defekt in der Osmoregulation hingewiesen.

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  1. Zoologisch-vergleichend-Anatomisohes Institut der Universität Zürich, Switzerland

    Martin Kaspar Rüegg

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Rüegg, M.K. Untersuchungen zum Proteinstoffwechsel des Wildtyps und der Letalmutante (ltr) von Drosophila melanogaster. Z. Vergl. Physiol. 60, 275–307 (1968). https://doi.org/10.1007/BF00298603

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