THE EFFECTS OF ELECTRICAL STIMULATION AND IONIC ALTERATIONS ON THE METABOLISM OF AMINO ACIDS AND PROTEINS IN EXCISED SUPERIOR CERVICAL GANGLIA OF THE RAT1

Authors

  • W. J. McBride,

    1. Department of Biochemistry, Schools of Medicine, Dentistry and Pharmacy, State University of New York at Buffalo, Buffalo, New York 14214
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    • 2

      Recipient of U.S. Public Health Service predoctoral Fellowship (1F1-GM-32, 889-02) NIGMS.

  • J. D. Klingman

    1. Department of Biochemistry, Schools of Medicine, Dentistry and Pharmacy, State University of New York at Buffalo, Buffalo, New York 14214
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  • 1

    Based in part on a dissertation submitted to the Graduate School, State University of New York at Buffalo in partial fulfillment of the requirements for the Ph.D. degree.

  • Present address: Section on Neurobiology, Institute of Psychiatric Research, Indiana University Medical Center, Indianapolis, Indiana 46202.

Abstract

Abstract— The effects of supramaximal electrical stimulation on the metabolism of amino acids and proteins in incubated superior cervical ganglia of the rat were studied by the use of a gas-liquid chromatographic (GLC) assay procedure. Stimulation at 5 Hz for 2 h caused an apparent increase in tissue levels of free amino acids, with alanine, serine, glycine, valine, threonine, isoleucine and aspartate (+ asparagine) most noticeably affected. The amino acid composition (partial) of the TCA-insoluble proteins of resting and stimulated ganglia was approximately the same after 60 min of incubation, but there was less TCA-insoluble protein in the stimulated ganglia. The addition of amino acids (at plasma concentrations) to the standard media had no apparent affect on the amino acid composition of this protein fraction. Stimulation for 0, 5 h initially increased the efflux of alanine, valine, proline and ornithine into the incubation media but prolonged stimulation (for 4–0 h) decreased the efflux of alanine, serine, glycine and isoleucine and increased the efflux of lysine into the incubation media. The leakage of amino acids from the ganglia appeared to be a sodium-dependent process.

The incorporation of 14C from [U-14C]glucose into glutamate (+ glutamine) and aspartate (+ asparagine) was greater in stimulated than in resting ganglia. However, the conversion of glutamate carbons from [U-14C]l-glutamate into aspartate was not affected by stimulation. Incorporation of 14C from [U-14C]glucose into glycine and serine was apparently not affected by stimulation during the 60 min of incubation. However, serine was the only amino acid which exhibited a higher specific radioactivity in stimulated ganglia than in resting ganglia incubated for 4 h in standard media. Lithium ions had the apparent specific effect of increasing the labelling with 14C from [U-14C]glucose into ornithine, and increasing the efflux and overall metabolism of serine in the ganglia. Incorporation of 14C from [U-14C]glucose into proteins was lower in the stimulated than in the resting ganglia if compensation was made for the higher radioactivity available in the total free amino acid pool of the stimulated ganglia.

The rate of 14C incorporation from [U-14C]glutamate into the TCA-insoluble proteins of resting ganglia was greater when no other amino acids at concentrations approximating plasma levels were added to the bathing media; this rate was lower in stimulated than in resting ganglia.

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