Optimal conditions for protein synthesis in incubated slices of rat brain.
Optimal conditions for protein synthesis in incubated slices of rat brain were determined to be: thickness 0.3 mm under air (for newborn 0.4-0.6 mm) or 0.5 mm under oxygen; temperature, 35-36 degrees C; hepes (N-2-hydroxyethylpiperazine-N'-2-ethane sulfonic acid) buffer; K+, 6-8 mM; Ca2+, 2-3 mM. Though maximum incorporation was found with a Na+ concentration of 110-12- mM. Though maximum incorporation was found with a Na+ concentration of 110-120mM, this requirement appears to be partly osmotic. The Na+ concentration may be reduced to 80 mM without inhibition of incorporation provided adjustment is made for osmotic balance. Mg2+. Optimal pH was 7.2-7.4. The rate of protein synthesis in this medium is 0.08-0.09% replacement of the protein amino acid/h in slices from adults and 1.6%/h in slices from 3-day-old rats. Thus slices from adults synthesize protein at 10-20% of the in vivo rate whereas slices at 3-day-old brainincorporate amino acid at 70-80% of the in vivo rate for young rats.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002021:Buffers; D002118:Calcium / Q000494:pharmacology; D003864:Depression, Chemical; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D012964:Sodium / Q000494:pharmacology; D013268:Stimulation, Chemical; D013696:Temperature
D000596:Amino Acids; D002021:Buffers; D009419:Nerve Tissue Proteins; D010042:Ouabain; D012964:Sodium; D008274:Magnesium; D011188:Potassium; D002118:Calcium