Quantitative measurement of cerebral protein synthesis in vivo: theory and methodological considerations.
The true rate of cerebral protein synthesis can be calculated from the ratio of labeled proteins to integrated arterial plasma amino acid specific activity (SA) only when the fraction of amino acid precursor pool dilution is known. In the following, current experimental designs on the measurement of cerebral protein synthesis are discussed and compared to our own approach in which the determination of regional precursor pool dilution by recycled unlabeled leucine is combined with the quantitation of regional cerebral protein synthesis rates. For this purpose, a constant arterial plasma leucine SA level is maintained for 45 min by programmed intravenous infusion which is sufficient for complete equilibrium between tissue leucine pool SAs and plasma free leucine SA. In addition to the regional assessment of the precursor dilution factor, protein radioactivity can be determined in the same tissue sample or in parallel brain sections of the same animal by quantitative autoradiography. It is then possible to calculate the actual rate of protein synthesis using the correct fraction of precursor pool dilution. This renders our approach particularly suitable for the quantitative measurement of regional CPS under pathological conditions.
Mies
G
G
Max-Planck-Institute for Neurological Research, Department of Experimental Neurology, Köln (Lindenthal), Germany. gmies@mpin-koeln.mpg.de
Djuricic
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B
Paschen
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Hossmann
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KA
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D006207:Half-Life; D007700:Kinetics; D007930:Leucine / Q000097:blood / Q000493:pharmacokinetics; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000096:biosynthesis*; D012356:RNA, Transfer, Leu / Q000378:metabolism; D051381:Rats
D000596:Amino Acids; D009419:Nerve Tissue Proteins; D012356:RNA, Transfer, Leu; D007930:Leucine