Skip to main content Accessibility help
×
×
Home

Distribution of vitamin C is tissue specific with early saturation of the brain and adrenal glands following differential oral dose regimens in guinea pigs

  • Stine Hasselholt (a1), Pernille Tveden-Nyborg (a1) and Jens Lykkesfeldt (a1)

Abstract

Vitamin C (VitC) deficiency is surprisingly common in humans even in developed parts of the world. The micronutrient has several established functions in the brain; however, the consequences of its deficiency are not well characterised. To elucidate the effects of VitC deficiency on the brain, increased knowledge about the distribution of VitC to the brain and within different brain regions after varying dietary concentrations is needed. In the present study, guinea pigs (like humans lacking the ability to synthesise VitC) were randomly divided into six groups (n 10) that received different concentrations of VitC ranging from 100 to 1500 mg/kg feed for 8 weeks, after which VitC concentrations in biological fluids and tissues were measured using HPLC. The distribution of VitC was found to be dynamic and dependent on dietary availability. Brain saturation was region specific, occurred at low dietary doses, and the dose–concentration relationship could be approximated with a three-parameter Hill equation. The correlation between plasma and brain concentrations of VitC was moderate compared with other organs, and during non-scorbutic VitC deficiency, the brain was able to maintain concentrations from about one-quarter to half of sufficient levels depending on the region, whereas concentrations in other tissues decreased to one-sixth or less. The adrenal glands have similar characteristics to the brain. The observed distribution kinetics with a low dietary dose needed for saturation and exceptional retention ability suggest that the brain and adrenal glands are high priority tissues with regard to the distribution of VitC.

Copyright

Corresponding author

* Corresponding author: Professor J. Lykkesfeldt, email jopl@sund.ku.dk

References

Hide All
1 US Department of Agriculture & Agricultural Research Service (2013) USDA National Nutrient Database for Standard Reference, Release 26. Nutrient Data Laboratory Home Page. http://www.ars.usda.gov/ba/bhnrc/ndl (accessed 18 March 2014).
2 Mosdol, A, Erens, B & Brunner, EJ (2008) Estimated prevalence and predictors of vitamin C deficiency within UK's low-income population. J Public Health (Oxf) 30, 456460.
3 Cahill, LE & El-Sohemy, A (2010) Haptoglobin genotype modifies the association between dietary vitamin C and serum ascorbic acid deficiency. Am J Clin Nutr 92, 14941500.
4 Schleicher, RL, Carroll, MD, Ford, ES, et al. (2009) Serum vitamin C and the prevalence of vitamin C deficiency in the United States: 2003–2004 National Health and Nutrition Examination Survey (NHANES). Am J Clin Nutr 90, 12521263.
5 Vitamin C Subcommittee of the Accessory Food Factors Committee, Medical Research Council (1948) Vitamin-C requirement of human adults; experimental study of vitamin-C deprivation in man. Lancet i, 853858.
6 Tveden-Nyborg, P, Johansen, LK, Raida, Z, et al. (2009) Vitamin C deficiency in early postnatal life impairs spatial memory and reduces the number of hippocampal neurons in guinea pigs. Am J Clin Nutr 90, 540546.
7 Myint, PK, Luben, RN, Welch, AA, et al. (2008) Plasma vitamin C concentrations predict risk of incident stroke over 10 y in 20 649 participants of the European Prospective Investigation into Cancer Norfolk prospective population study. Am J Clin Nutr 87, 6469.
8 Higdon, J & Angelo, Gu (2014) Vitamin C. The Linus Pauling Institute Micronutrient Information Center (MIC). http://lpi.oregonstate.edu/infocenter/vitamins/vitaminC/ (accessed 27 April 2014).
9 Tveden-Nyborg, P & Lykkesfeldt, J (2013) Does vitamin C deficiency increase lifestyle-associated vascular disease progression? Evidence based on experimental and clinical studies. Antioxid Redox Signal 19, 20842104.
10 Lykkesfeldt, J, Trueba, GP, Poulsen, HE, et al. (2007) Vitamin C deficiency in weanling guinea pigs: differential expression of oxidative stress and DNA repair in liver and brain. Br J Nutr 98, 11161119.
11 Hughes, RE, Hurley, RJ & Jones, PR (1971) The retention of ascorbic acid by guinea-pig tissues. Br J Nutr 26, 433438.
12 Harrison, FE, Dawes, SM, Meredith, ME, et al. (2010) Low vitamin C and increased oxidative stress and cell death in mice that lack the sodium-dependent vitamin C transporter SVCT2. Free Radic Biol Med 49, 821829.
13 Reiber, H, Martens, U, Prall, F, et al. (1994) Relevance of endogenous ascorbate and tocopherol for brain cell vitality indicated by photon emission. J Neurochem 62, 608614.
14 Tanaka, K, Hashimoto, T, Tokumaru, S, et al. (1997) Interactions between vitamin C and vitamin E are observed in tissues of inherently scorbutic rats. J Nutr 127, 20602064.
15 Hara, K & Akiyama, Y (2009) Collagen-related abnormalities, reduction in bone quality, and effects of menatetrenone in rats with a congenital ascorbic acid deficiency. J Bone Miner Metab 27, 324332.
16 Walmsley, AR, Batten, MR, Lad, U, et al. (1999) Intracellular retention of procollagen within the endoplasmic reticulum is mediated by prolyl 4-hydroxylase. J Biol Chem 274, 1488414892.
17 Yoshikawa, K, Takahashi, S, Imamura, Y, et al. (2001) Secretion of non-helical collagenous polypeptides of α1(IV) and α2(IV) chains upon depletion of ascorbate by cultured human cells. J Biochem 129, 929936.
18 Sotiriou, S, Gispert, S, Cheng, J, et al. (2002) Ascorbic-acid transporter Slc23a1 is essential for vitamin C transport into the brain and for perinatal survival. Nat Med 8, 514517.
19 Tomita, S, Ueno, M, Sakamoto, M, et al. (2003) Defective brain development in mice lacking the Hif-1α gene in neural cells. Mol Cell Biol 23, 67396749.
20 Flashman, E, Davies, SL, Yeoh, KK, et al. (2010) Investigating the dependence of the hypoxia-inducible factor hydroxylases (factor inhibiting HIF and prolyl hydroxylase domain 2) on ascorbate and other reducing agents. Biochem J 427, 135142.
21 Vissers, MC, Gunningham, SP, Morrison, MJ, et al. (2007) Modulation of hypoxia-inducible factor-1 α in cultured primary cells by intracellular ascorbate. Free Radic Biol Med 42, 765772.
22 Kuiper, C, Dachs, GU, Currie, MJ, et al. (2014) Intracellular ascorbate enhances hypoxia-inducible factor (HIF)-hydroxylase activity and preferentially suppresses the HIF-1 transcriptional response. Free Radic Biol Med 69, 308317.
23 Diliberto, EJ Jr & Allen, PL (1980) Semidehydroascorbate as a product of the enzymic conversion of dopamine to norepinephrine. Coupling of semidehydroascorbate reductase to dopamine-β-hydroxylase. Mol Pharmacol 17, 421426.
24 Diliberto, EJ Jr & Allen, PL (1981) Mechanism of dopamine-β-hydroxylation. Semidehydroascorbate as the enzyme oxidation product of ascorbate. J Biol Chem 256, 33853393.
25 Levine, M, Morita, K, Heldman, E, et al. (1985) Ascorbic acid regulation of norepinephrine biosynthesis in isolated chromaffin granules from bovine adrenal medulla. J Biol Chem 260, 1559815603.
26 May, JM, Qu, ZC, Nazarewicz, R, et al. (2013) Ascorbic acid efficiently enhances neuronal synthesis of norepinephrine from dopamine. Brain Res Bull 90, 3542.
27 Meredith, ME & May, JM (2013) Regulation of embryonic neurotransmitter and tyrosine hydroxylase protein levels by ascorbic acid. Brain Res 1539, 714.
28 Lane, DJ & Lawen, A (2013) The glutamate aspartate transporter (GLAST) mediates l-glutamate-stimulated ascorbate-release via swelling-activated anion channels in cultured neonatal rodent astrocytes. Cell Biochem Biophys 65, 107119.
29 Cammack, J, Ghasemzadeh, B & Adams, RN (1991) The pharmacological profile of glutamate-evoked ascorbic acid efflux measured by in vivo electrochemistry. Brain Res 565, 1722.
30 Heller, R, Unbehaun, A, Schellenberg, B, et al. (2001) l-Ascorbic acid potentiates endothelial nitric oxide synthesis via a chemical stabilization of tetrahydrobiopterin. J Biol Chem 276, 4047.
31 Huang, A, Vita, JA, Venema, RC, et al. (2000) Ascorbic acid enhances endothelial nitric-oxide synthase activity by increasing intracellular tetrahydrobiopterin. J Biol Chem 275, 1739917406.
32 Baker, TA, Milstien, S & Katusic, ZS (2001) Effect of vitamin C on the availability of tetrahydrobiopterin in human endothelial cells. J Cardiovasc Pharmacol 37, 333338.
33 Ward, MS, Lamb, J, May, JM, et al. (2013) Behavioral and monoamine changes following severe vitamin C deficiency. J Neurochem 124, 363375.
34 Harrison, FE, Bowman, GL & Polidori, MC (2014) Ascorbic acid and the brain: rationale for the use against cognitive decline. Nutrients 6, 17521781.
35 Tsukaguchi, H, Tokui, T, Mackenzie, B, et al. (1999) A family of mammalian Na+-dependent l-ascorbic acid transporters. Nature 399, 7075.
36 Wang, H, Dutta, B, Huang, W, et al. (1999) Human Na(+)-dependent vitamin C transporter 1 (hSVCT1): primary structure, functional characteristics and evidence for a non-functional splice variant. Biochim Biophys Acta 1461, 19.
37 Daruwala, R, Song, J, Koh, WS, et al. (1999) Cloning and functional characterization of the human sodium-dependent vitamin C transporters hSVCT1 and hSVCT2. FEBS Lett 460, 480484.
38 Godoy, A, Ormazabal, V, Moraga-Cid, G, et al. (2007) Mechanistic insights and functional determinants of the transport cycle of the ascorbic acid transporter SVCT2. Activation by sodium and absolute dependence on bivalent cations. J Biol Chem 282, 615624.
39 Rumsey, SC, Kwon, O, Xu, GW, et al. (1997) Glucose transporter isoforms GLUT1 and GLUT3 transport dehydroascorbic acid. J Biol Chem 272, 1898218989.
40 Ulloa, V, Garcia-Robles, M, Martinez, F, et al. (2013) Human choroid plexus papilloma cells efficiently transport glucose and vitamin C. J Neurochem 127, 403414.
41 Agus, DB, Gambhir, SS, Pardridge, WM, et al. (1997) Vitamin C crosses the blood–brain barrier in the oxidized form through the glucose transporters. J Clin Invest 100, 28422848.
42 Lindblad, M, Tveden-Nyborg, P & Lykkesfeldt, J (2013) Regulation of vitamin C homeostasis during deficiency. Nutrients 5, 28602879.
43 Lykkesfeldt, J & Poulsen, HE (2010) Is vitamin C supplementation beneficial? Lessons learned from randomised controlled trials. Br J Nutr 103, 12511259.
44 Frei, B, Birlouez-Aragon, I & Lykkesfeldt, J (2012) Authors' perspective: what is the optimum intake of vitamin C in humans? Crit Rev Food Sci Nutr 52, 815829.
45 EFSA Panel on Dietetic Products, Nutrition and Allergies (2013) Scientific Opinion on Dietary Reference Values for vitamin C. EFSA J 11, 68.
46 Burns, JJ & Evans, C (1956) The synthesis of l-ascorbic acid in the rat from d-glucuronolactone and l-gulonolactone. J Biol Chem 223, 897905.
47 Burns, JJ (1957) Missing step in man, monkey and guinea pig required for the biosynthesis of l-ascorbic acid. Nature 180, 553.
48 Burns, JJ, Peyser, P & Moltz, A (1956) Missing step in guinea pigs required for the biosynthesis of l-ascorbic acid. Science 124, 11481149.
49 Nishikimi, M, Kawai, T & Yagi, K (1992) Guinea pigs possess a highly mutated gene for l-gulono-γ-lactone oxidase, the key enzyme for l-ascorbic acid biosynthesis missing in this species. J Biol Chem 267, 2196721972.
50 Nishikimi, M, Fukuyama, R, Minoshima, S, et al. (1994) Cloning and chromosomal mapping of the human nonfunctional gene for l-gulono-γ-lactone oxidase, the enzyme for l-ascorbic acid biosynthesis missing in man. J Biol Chem 269, 1368513688.
51 Berger, J, Shepard, D, Morrow, F, et al. (1989) Relationship between dietary intake and tissue levels of reduced and total vitamin C in the nonscorbutic guinea pig. J Nutr 119, 734740.
52 Tveden-Nyborg, P, Vogt, L, Schjoldager, JG, et al. (2012) Maternal vitamin C deficiency during pregnancy persistently impairs hippocampal neurogenesis in offspring of guinea pigs. PLOS ONE 7, e48488.
53 Lykkesfeldt, J (2012) Ascorbate and dehydroascorbic acid as biomarkers of oxidative stress: validity of clinical data depends on vacutainer system used. Nutr Res 32, 6669.
54 Lykkesfeldt, J, Loft, S & Poulsen, HE (1995) Determination of ascorbic acid and dehydroascorbic acid in plasma by high-performance liquid chromatography with coulometric detection – are they reliable biomarkers of oxidative stress? Anal Biochem 229, 329335.
55 Lykkesfeldt, J (2002) Measurement of ascorbic acid and dehydroascorbic acid in biological samples. Curr Protoc Toxicol Chapter 7, 7·6·17·615.
56 Lykkesfeldt, J (2000) Determination of ascorbic acid and dehydroascorbic acid in biological samples by high-performance liquid chromatography using subtraction methods: reliable reduction with tris[2-carboxyethyl]phosphine hydrochloride. Anal Biochem 282, 8993.
57 Lykkesfeldt, J (2007) Ascorbate and dehydroascorbic acid as reliable biomarkers of oxidative stress: analytical reproducibility and long-term stability of plasma samples subjected to acidic deproteinization. Cancer Epidemiol Biomarkers Prev 16, 25132516.
58 Mortensen, A, Hasselholt, S, Tveden-Nyborg, P, et al. (2013) Guinea pig ascorbate status predicts tetrahydrobiopterin plasma concentration and oxidation ratio in vivo . Nutr Res 33, 859867.
59 Mun, GH, Kim, MJ, Lee, JH, et al. (2006) Immunohistochemical study of the distribution of sodium-dependent vitamin C transporters in adult rat brain. J Neurosci Res 83, 919928.
60 Harrison, FE, Green, RJ, Dawes, SM, et al. (2010) Vitamin C distribution and retention in the mouse brain. Brain Res 1348, 181186.
61 Bahney, J & von Bartheld, CS (2014) Validation of the isotropic fractionator: comparison with unbiased stereology and DNA extraction for quantification of glial cells. J Neurosci Methods 222, 165174.
62 Herculano-Houzel, S & Lent, R (2005) Isotropic fractionator: a simple, rapid method for the quantification of total cell and neuron numbers in the brain. J Neurosci 25, 25182521.
63 Rice, ME & Russo-Menna, I (1998) Differential compartmentalization of brain ascorbate and glutathione between neurons and glia. Neuroscience 82, 12131223.
64 Berger, UV & Hediger, MA (2000) The vitamin C transporter SVCT2 is expressed by astrocytes in culture but not in situ . Neuroreport 11, 13951399.
65 Mefford, IN, Oke, AF & Adams, RN (1981) Regional distribution of ascorbate in human brain. Brain Res 212, 223226.
66 Frikke-Schmidt, H, Tveden-Nyborg, P, Birck, MM, et al. (2011) High dietary fat and cholesterol exacerbates chronic vitamin C deficiency in guinea pigs. Br J Nutr 105, 5461.
67 Reiber, H, Ruff, M & Uhr, M (1993) Ascorbate concentration in human cerebrospinal fluid (CSF) and serum. Intrathecal accumulation and CSF flow rate. Clin Chim Acta 217, 163173.
68 Tallaksen, CM, Bohmer, T & Bell, H (1992) Concentrations of the water-soluble vitamins thiamin, ascorbic acid, and folic acid in serum and cerebrospinal fluid of healthy individuals. Am J Clin Nutr 56, 559564.
69 Angelow, S, Haselbach, M & Galla, HJ (2003) Functional characterisation of the active ascorbic acid transport into cerebrospinal fluid using primary cultured choroid plexus cells. Brain Res 988, 105113.
70 Vannucci, SJ (1994) Developmental expression of GLUT1 and GLUT3 glucose transporters in rat brain. J Neurochem 62, 240246.
71 Kuehne, LK, Reiber, H, Bechter, K, et al. (2013) Cerebrospinal fluid neopterin is brain-derived and not associated with blood–CSF barrier dysfunction in non-inflammatory affective and schizophrenic spectrum disorders. J Psychiatr Res 47, 14171422.
72 Søgaard, D, Lindblad, MM, Paidi, MD, et al. (2014) In vivo vitamin C deficiency in guinea pigs increases ascorbate transporters in liver but not kidney and brain Nutr Res 34, 639645.
73 Bornstein, SR, Yoshida-Hiroi, M, Sotiriou, S, et al. (2003) Impaired adrenal catecholamine system function in mice with deficiency of the ascorbic acid transporter (SVCT2). FASEB J 17, 19281930.
74 Lee, JH, Oh, CS, Mun, GH, et al. (2006) Immunohistochemical localization of sodium-dependent l-ascorbic acid transporter 1 protein in rat kidney. Histochem Cell Biol 126, 491494.
75 Corpe, CP, Tu, H, Eck, P, et al. (2010) Vitamin C transporter Slc23a1 links renal reabsorption, vitamin C tissue accumulation, and perinatal survival in mice. J Clin Invest 120, 10691083.
76 Levine, M, Conry-Cantilena, C, Wang, Y, et al. (1996) Vitamin C pharmacokinetics in healthy volunteers: evidence for a recommended dietary allowance. Proc Natl Acad Sci U S A 93, 37043709.
77 Levine, M, Wang, Y, Padayatty, SJ, et al. (2001) A new recommended dietary allowance of vitamin C for healthy young women. Proc Natl Acad Sci U S A 98, 98429846.
78 Macias, RI, Hierro, C, de Juan, SC, et al. (2011) Hepatic expression of sodium-dependent vitamin C transporters: ontogeny, subtissular distribution and effect of chronic liver diseases. Br J Nutr 106, 18141825.
79 Savini, I, Rossi, A, Pierro, C, et al. (2008) SVCT1 and SVCT2: key proteins for vitamin C uptake. Amino Acids 34, 347355.
80 Reidling, JC & Rubin, SA (2011) Promoter analysis of the human ascorbic acid transporters SVCT1 and 2: mechanisms of adaptive regulation in liver epithelial cells. J Nutr Biochem 22, 344350.
81 Meredith, ME, Harrison, FE & May, JM (2011) Differential regulation of the ascorbic acid transporter SVCT2 during development and in response to ascorbic acid depletion. Biochem Biophys Res Commun 414, 737742.
82 Loria, CM, Whelton, PK, Caulfield, LE, et al. (1998) Agreement among indicators of vitamin C status. Am J Epidemiol 147, 587596.
83 Timpson, NJ, Forouhi, NG, Brion, MJ, et al. (2010) Genetic variation at the SLC23A1 locus is associated with circulating concentrations of l-ascorbic acid (vitamin C): evidence from 5 independent studies with >15,000 participants. Am J Clin Nutr 92, 375382.
84 Eck, P, Erichsen, HC, Taylor, JG, et al. (2004) Comparison of the genomic structure and variation in the two human sodium-dependent vitamin C transporters, SLC23A1 and SLC23A2. Hum Genet 115, 285294.
85 Michels, AJ, Hagen, TM & Frei, B (2013) Human genetic variation influences vitamin C homeostasis by altering vitamin C transport and antioxidant enzyme function. Annu Rev Nutr 33, 4570.
86 Williams, BH (2012) Chapter 24 – non-infectious diseases. In The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents, pp. 685704 [Suckow, MA, Stevens, KA and Wilson, RP, editors]. Boston, MA: Academic Press.
87 Maeda, N, Hagihara, H, Nakata, Y, et al. (2000) Aortic wall damage in mice unable to synthesize ascorbic acid. Proc Natl Acad Sci U S A 97, 841846.
88 Kawai, T, Nishikimi, M, Ozawa, T, et al. (1992) A missense mutation of l-gulono-γ-lactone oxidase causes the inability of scurvy-prone osteogenic disorder rats to synthesize l-ascorbic acid. J Biol Chem 267, 2197321976.
89 Mizushima, Y, Harauchi, T, Yoshizaki, T, et al. (1984) A rat mutant unable to synthesize vitamin C. Experientia 40, 359361.
90 Ogiri, Y, Sun, F, Hayami, S, et al. (2002) Very low vitamin C activity of orally administered l-dehydroascorbic acid. J Agric Food Chem 50, 227229.
91 Cui, Y, Otsuka, M & Fujiwara, Y (2001) Reduction of dehydroerythorbic acid in vitamin C-deficient guinea pigs. J Nutr Sci Vitaminol (Tokyo) 47, 316320.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

British Journal of Nutrition
  • ISSN: 0007-1145
  • EISSN: 1475-2662
  • URL: /core/journals/british-journal-of-nutrition
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed