Subcellular localization of dopamine β-hydroxylase and endogenous norepinephrine in the rat hypothalamus

doi:10.1016/0006-8993(74)90236-4

Brain Research

Volume 65, Issue 3, 18 January 1974, Pages 475-487

https://doi.org/10.1016/0006-8993(74)90236-4 Get rights and content

Abstract

The subcellular distribution of endogenous norepinephrine, dopamine β-hydroxylase (EC 1.14.2.1) and endogenous inhibitors to dopamine β-hydroxylase have been examined in the rat hypothalamus. Sixty percent of the recovered norepinephrine and activity of dopamine β-hydroxylase sedimented in the crude mitochondrial (P₂) fraction; fractionation on a linear sucrose gradient revealed that the particulate-bound amine and enzyme were localized in the synaptosomal fractions. After osmotic shock of the P₂ fraction the released storage vesicles were purified by differential centrifugation or centrifugation on a sucrose gradient; 40–70% of the dopamine β-hydroxylase activity released from the P₂ fraction was localized in the fractions reported to contain storage vesicles. In contrast, less than 10% of P₂-norephinephrine was found in these fractions. The fraction from the gradient that had the highest specific activity of dopamine β-hydroxylase was shown by electron microscopy to be markedly enriched with large, dense-core vesicles. Endogenous inhibitors to dopamine β-hydroxylase were found in all subcellular fractions with only a 2.5-fold difference between the least and most inhibitory fractions.

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Subcellular localization of dopamine β-hydroxylase and endogenous norepinephrine in the rat hypothalamus

Abstract

Int. Rev. Neurobiol.

Biochem. Pharmacol.

Tissue and Cell

Life Sci.

Biochem. Pharmacol.

Biochem. Pharmacol.

J. biol. Chem.

Biochem. Pharmacol.

Tissue and Cell

Observations on the localization of noradrenaline in homogenates of dog's hypothalamus

Evidence for two types of noradrenaline storage particles in dog spleen

Nature (Lond.)

Dopamine-β-hydroxylase in rat brain: developmental characteristics

J. Neurochem.

Catecholamines in fetal and newborn rat brain

J. Neurochem.

Catecholamine uptake by synaptosomes in homogenates of rat brain: stereospecificity in different areas

J. Pharmacol. exp. Ther.