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Effect of ascorbic acid deficiency on catecholamine synthesis in adrenal glands of SMP30/GNL knockout mice

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Abstract

Purpose

The effect of an AA deficiency on catecholamine biosynthesis in adult mice in vivo is unknown. Therefore, we quantified catecholamine and the expression of catecholamine synthetic enzymes in the adrenal glands of senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice placed in an AA-deficient state.

Methods

At 30 days of age, mice were divided into the following 4 groups: AA (−) SMP30/GNL KO, AA (+) SMP30/GNL KO, AA (−) wild type (WT), and AA (+) WT. The AA (+) groups were given water containing 1.5 g/L AA, whereas the AA (−) groups received water without AA until the experiment ended. In addition, all mice were fed an AA-depleted diet. Catecholamine levels were measured by a liquid chromatographic method. Tyrosine hydroxylase, dopa decarboxylase, dopamine β-hydroxylase, and phenylethanolamine N-methyltransferase mRNA expression levels were measured with the quantitative real-time polymerase chain reaction (qPCR). Tyrosine hydroxylase and dopamine β-hydroxylase protein levels were quantified by Western blot analysis.

Results

In the adrenals of AA (−) SMP30/GNL KO mice, noradrenaline and adrenaline levels decreased significantly compared to other three groups of mice, although there were no significant differences in dopamine β-hydroxylase or phenylethanolamine N-methyltransferase mRNA content. Moreover, there was no significant difference in their dopamine β-hydroxylase protein levels. On the other hand, AA depletion did not affect dopamine levels in adrenal glands of mice.

Conclusion

An AA deficiency decreases the noradrenaline and adrenaline levels in adrenal glands of mice in vivo.

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Abbreviations

AA:

Ascorbic acid

AD:

Adrenaline

ANOVA:

Analysis of variance

BH3 · radical:

Trihydrobiopterin radical

BH4 :

Tetrahydrobiopterin

cDNA:

Complementary DNA

Cu+ :

Cuprous ion

Ct:

Threshold cycle

DA:

Dopamine

DBH:

Dopamine β-hydroxylase

DDC:

Dopa decarboxylase

ECD:

Electrochemical detector

EDTA:

Ethylenediaminetetraacetic acid

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

GNL:

Gluconolactonase

HPLC:

High-performance liquid chromatography

KO:

Knockout

NA:

Noradrenaline

PNMT:

Phenylethanolamine N-methyltransferase

qPCR:

Quantitative real-time polymerase chain reaction

RGN:

Regucalcin

SMP30:

Senescence marker protein-30

SVCT:

Sodium-dependent vitamin C transporter

TH:

Tyrosine hydroxylase

WT:

Wild type

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Acknowledgments

The authors thank Ms. Phyllis Minick for the excellent English editorial assistance. Vitamin C powder was kindly provided by DSM Nutrition Japan. This study is supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number 24380073 (A.I.) and 23590441 (N.M.) and by JSPS Research Fellowship for Young Scientists Grant Number 00234778 (A.A.).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Molecular Regulation of Aging, Tokyo Metropolitan Institute of Gerontology (TMIG), 35-2 Sakae-cho, Itabashi-ku, Tokyo, 173-0015, Japan

    Akiko Amano, Naoki Maruyama & Akihito Ishigami

  2. Cellular Genetics, Graduate School of Science and Engineering, Tokyo Metropolitan University, Tokyo, 192-0397, Japan

    Akiko Amano & Toshiro Aigaki

  3. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, 113-0033, Japan

    Makoto Tsunoda

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  1. Akiko Amano
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Correspondence to Akihito Ishigami.

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Amano, A., Tsunoda, M., Aigaki, T. et al. Effect of ascorbic acid deficiency on catecholamine synthesis in adrenal glands of SMP30/GNL knockout mice. Eur J Nutr 53, 177–185 (2014). https://doi.org/10.1007/s00394-013-0515-9

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