An analysis of nucleotides and catecholamines in bovine medullary granules by anion exchange high pressure liquid chromatography and fluorescence: Evidence that most of the catecholamines in chromaffin granules are stored without associated atp

Knox Van Dyke, Robert Robinson, Pedro Urquilla, David Smith, Michael Taylor, Michael Trush, Mark Wilson

Research output: Contribution to journalArticle

Abstract

The catecholamine and nucleotide content of chromaffin granules from bovine adrenal medulla was analyzed both qualitatively and quantitatively. Nucleotides were examined by separation on anion exchange high pressure liquid chromatography and ultraviolet absorption, as well as by anion exchange thin-layer chromatography and luciferin-luciferase light emission analysis. Catecholamines were quantified via automated contin-uous-flow analysis using fluorescence. We found the average catecholamine/ATP ratio to be 8.2:1 with an SD of 1.98 and an SE of 0.50 for 16 separate glands. Relative percentages of epinephrine and norepinephrine were found to be 79.8 and 20.2, respectively. The granules contained 80.9% ATP, 12.3% GTP, and 6.8% UTP, as well as detectable amounts of ADP, AMP, GDP, and UDP. If we accept the catecholamine/ATP ratio of 4:1 proposed by Hillarp earlier, then those catecholamines constituting the remainder of our ratio of 8.2:1 should represent a nucleotide-unassociated pool. In view of recent evidence of a direct agonistic function of nucleotides, we propose that the granule-associated nucleotides may act locally as coagonists with certain biogenic amines, and may additionally provide a circulating pool of purines and pyrimidines for use by the heart and lungs.

Original languageEnglish (US)
Pages (from-to)377-391
Number of pages15
JournalPharmacology
Volume15
Issue number5
DOIs
StatePublished - 1977
Externally publishedYes

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Chromaffin Granules
Catecholamines
Anions
Nucleotides
Fluorescence
High Pressure Liquid Chromatography
Adenosine Triphosphate
Pyrimidines
Purines
Uridine Triphosphate
Adrenal Medulla
Biogenic Amines
Uridine Diphosphate
Adenosine Monophosphate
Thin Layer Chromatography
Guanosine Triphosphate
Luciferases
Adenosine Diphosphate
Epinephrine
Norepinephrine

Keywords

  • Adenosine triphosphate
  • Adrenal Bovine Catecholamines
  • Epinephrine
  • Granules
  • Guanosine triphosphate
  • High pressure liquid chromatography
  • Norepinephrine
  • Nucleotides
  • Uridine triphosphate

ASJC Scopus subject areas

  • Pharmacology

Cite this

An analysis of nucleotides and catecholamines in bovine medullary granules by anion exchange high pressure liquid chromatography and fluorescence : Evidence that most of the catecholamines in chromaffin granules are stored without associated atp. / Van Dyke, Knox; Robinson, Robert; Urquilla, Pedro; Smith, David; Taylor, Michael; Trush, Michael; Wilson, Mark.

In: Pharmacology, Vol. 15, No. 5, 1977, p. 377-391.

Research output: Contribution to journalArticle

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abstract = "The catecholamine and nucleotide content of chromaffin granules from bovine adrenal medulla was analyzed both qualitatively and quantitatively. Nucleotides were examined by separation on anion exchange high pressure liquid chromatography and ultraviolet absorption, as well as by anion exchange thin-layer chromatography and luciferin-luciferase light emission analysis. Catecholamines were quantified via automated contin-uous-flow analysis using fluorescence. We found the average catecholamine/ATP ratio to be 8.2:1 with an SD of 1.98 and an SE of 0.50 for 16 separate glands. Relative percentages of epinephrine and norepinephrine were found to be 79.8 and 20.2, respectively. The granules contained 80.9{\%} ATP, 12.3{\%} GTP, and 6.8{\%} UTP, as well as detectable amounts of ADP, AMP, GDP, and UDP. If we accept the catecholamine/ATP ratio of 4:1 proposed by Hillarp earlier, then those catecholamines constituting the remainder of our ratio of 8.2:1 should represent a nucleotide-unassociated pool. In view of recent evidence of a direct agonistic function of nucleotides, we propose that the granule-associated nucleotides may act locally as coagonists with certain biogenic amines, and may additionally provide a circulating pool of purines and pyrimidines for use by the heart and lungs.",
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