Acceleration of noradrenaline biosynthesis in the guinea-pig vas deferens by potassium.
1. Increasing the concentration of KCl in Krebs-Henseleit bicarbonate solution enhanced the formation of (14)C-noradrenaline ((14)C-NA) from (14)C-tyrosine in the guinea-pig vas deferens. In 52 mM KCl Krebs-Henseleit solution the specific activity of the newly formed (14)C-NA was double that of controls.2. The rate of synthesis of (14)C-NA from (14)C-tyrosine was constant for up to 2 h in 52 mM KCl Krebs-Henseleit solution and for 4 h in unmodified Krebs-Henseleit solution.3. There was no increase in NA formation in the presence of KCl rich Krebs-Henseleit solution if (14)C-DOPA was used as the starting substrate instead of (14)C-tyrosine.4. The specific activity of (14)C-tyrosine in the high KCl treated vas deferens was 80% of that of control tissues. Thus the enhanced synthesis of (14)C-NA in high KCl Krebs-Henseleit solution did not arise from an increase in the specific activity of precursor.5. The effect of K(+) on NA synthesis was not mimicked by ganglionic stimulants nor blocked by tetrodotoxin.6. Removal of Ca(2+) ions or increasing the concentration of Mg(2+) ions abolished the increase in synthesis of NA seen in high KCl Krebs-Henseleit solution but left the basal rate of NA synthesis in unmodified Krebs-Henseleit solution unaltered.7. The spontaneous release of newly synthesized catecholamines ((14)C-labelled) or tritiated noradrenaline ((3)H-NA) from vasa deferentia was increased in 52 mM KCl Krebs-Henseleit solution. Removal of Ca(2+) ions reduced the increased efflux of newly synthesized amine in high KCl media to that seen in unmodified Krebs-Henseleit solution. The efflux of (3)H-NA was reduced to one-third of its former rate in the absence of Ca(2+).8. High KCl Krebs-Henseleit solution caused a substantial contraction of the vas deferens which was not abolished by tetrodotoxin. Release of (3)H-NA paralleled the contractile response, and was likewise unaffected by tetrodotoxin.9. No evidence was obtained for any alterations in the activity of tyrosine hydroxylase, the rate limiting enzyme in the formation of NA from tyrosine, in homogenates of vas deferens which had been treated with 52 mM KCl Krebs-Henseleit solution.10. These results support the hypothesis that acceleration of NA synthesis occurs when tyrosine hydroxylase is freed from end-product inhibition by the release of noradrenaline, brought about in this case, by high concentrations of KCl.
D000818:Animals; D001337:Autonomic Agents / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002247:Carbon Isotopes; D002845:Chromatography; D002852:Chromatography, Ion Exchange; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D006899:Mixed Function Oxygenases; D009638:Norepinephrine / Q000096:biosynthesis*; D011188:Potassium / Q000494:pharmacology*; D013779:Tetrodotoxin / Q000494:pharmacology; D014316:Tritium; D014443:Tyrosine / Q000378:metabolism; D014649:Vas Deferens / Q000187:drug effects* / Q000378:metabolism
D001337:Autonomic Agents; D002247:Carbon Isotopes; D014316:Tritium; D014443:Tyrosine; D013779:Tetrodotoxin; D006899:Mixed Function Oxygenases; D008274:Magnesium; D011188:Potassium; D002118:Calcium; D009638:Norepinephrine