Biochemical and autoradiographical determination of protein synthesis in experimental brain tumors of rats.
The rate of leucine incorporation into brain proteins was studied in rats with experimental brain tumors produced by intracerebral transplantation of the glioma clone F98. Incorporation was measured with [14C]leucine using a controlled infusion technique for maintaining constant specific activity of [14C]leucine in plasma, followed by quantitative autoradiography and biochemical tissue analysis. After 45 min the specific activity of free [14C]leucine in plasma was 2.5-3 times higher than in brain and brain tumor, indicating that the precursor pool for protein synthesis was fueled both by exogenous (plasma-derived) and endogenous (proteolysis-derived) amino acids. Endogenous recycling of amino acids amounted to 73% of total free leucine pool in brain tumors and to 60-70% in normal brain. Taking endogenous amino acid recycling into account, leucine incorporation was 78.7 +/- 16.0 nmol/g of tissue/min in brain tumor, and 17.2 +/- 4.2 and 9.7 +/- 3.3 nmol/g/min in normal frontal cortex and striatum, respectively. Leucine incorporation within tumor tissue was markedly heterogeneous, depending on the local pattern of tumor proliferation and necrosis. Our results demonstrate that quantitative measurement of leucine incorporation into brain proteins requires estimation of recycling of amino acids derived from proteolysis and, in consequence, biochemical determination of the free amino acid precursor pool in tissue samples. With the present approach such measurements are possible and provide the quantitative basis for the evaluation of therapeutic interventions.
Widmann
R
R
Department of Experimental Neurology, Max-Planck-Institute for Neurological Research, Cologne, F.R.G.
Kocher
M
M
Ernestus
R I
RI
Hossmann
K A
KA
D000818:Animals; D001345:Autoradiography; D001932:Brain Neoplasms / Q000097:blood / Q000378:metabolism*; D007930:Leucine / Q000097:blood / Q000378:metabolism; D008297:Male; D009363:Neoplasm Proteins / Q000096:biosynthesis*; D051381:Rats; D011919:Rats, Inbred Strains
D009363:Neoplasm Proteins; D007930:Leucine