In vitro trehalose synthesis in American cockroach fat body measured by NMR spectroscopy using ^<13>C-sugars and a trehalase inhibitor, validoxylamine A

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  • ^<13>C ラベル前駆体とトレハラーゼ阻害剤を用いた, In vitro でのゴキブリ脂肪体のトレハロース合成能の NMR による測定

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Abstract

Synthesis of trehalose in cultured tissues and organs of the American cockroach, Periplaneta americana, and effect of a trehalase inhibitor, validoxylamine A (VAA), on it were studied using a medium containing ^<13>C-labelled precursors, [^<13>C-1] glucose or [^<13>C-1] fructose by ^<13>C-NMR spectroscopy. Trehalose was produced only by the fat body among the tested tissues and organs. The abdominal fat body utilized glucose and fructose as an energy source as well as a material for trehalose synthesis in vitro. VAA enhanced the trehalose synthesis both in vivo and in vitro, even when the fat body used for the culture was obtained from a cockroach previously injected with VAA. The fat body from the female cockroach had the capacity to produce about 212 nmol of trehalose/hr/insect in a medium with 14mM [^<13>C-1] glucose and 10^<-6>M VAA. When fructose was used as a precursor, the fat body produced glucose as well as trehalose, but trehalose only in the presence of VAA. Feedback control of trehalose synthesis by trehalose was observed in the cultured fat body with fructose as a precursor. These results indicated that the fat body hydrolyzed trehalose along with the trehalose synthesis, and that VAA had no effect on the trehalose synthesis itself. ^<13>C-NMR with the tissue culture system containing [^<13>C-1] precursor and VAA was proved to give satisfactory results for estimating the synthetic activity of trehalose and sugar metabolism in the insect fat body.

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