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Multicompartment analysis of protein-restricted phenylketonuric mice reveals amino acid imbalances in brain

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Background

The mainstay of therapy for phenylketonuria (PKU) remains dietary protein restriction. Developmental and neurocognitive outcomes for patients, however, remain suboptimal. We tested the hypothesis that mice with PKU receiving protein-restricted diets would reveal disruptions of brain amino acids that shed light on these neurocognitive deficits.

Method

Phenylalanine hydroxylase-deficient (PKU) mice and parallel controls (both wild-type and heterozygous) were fed custom diets containing 18, 6, and 4 % protein for 3 weeks, after which tissues (brain, liver, sera) were collected for amino acid analysis profiling.

Results

Phenylalanine (phe) was increased in all tissues (p < 0.0001) of PKU mice and improved with protein restriction. In sera, decreased tyrosine (p < 0.01) was corrected (defined as not significantly different from the level in control mice receiving 18 % chow) with protein restriction, whereas protein restriction significantly increased many other amino acids. A similar trend for increased amino acid levels with protein restriction was also observed in liver. In brain, the effects of protein restriction on large neutral amino acids (LNAAs) were variable, with some deficit correction (threonine, methionine, glutamine) and no correction of tyrosine under any dietary paradigm. Protein restriction (4 % diet) in PKU mice significantly decreased lysine, arginine, taurine, glutamate, asparagine, and serine which had been comparable to control mice under 18 % protein intake.

Conclusion

Depletion of taurine, glutamate, and serine in the brain of PKU mice with dietary protein restriction may provide new insight into neurocognitive deficits of PKU.

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Abbreviations

Phe:

Phenylalanine

tyr:

Tyrosine

trp:

Tryptophan

lys:

Lysine

arg:

Arginine

glu:

Glutamate

asp:

Aspartic acid

val:

Valine

ile:

Isoleucine

leu:

Leucine

thr:

Threonine

met:

Methionine

gln:

Glutamine

his:

Histidine

gly:

Glycine

pro:

Proline

ala:

Alanine

β-ala:

β-alanine

ser:

Serine

cit:

Citrulline

tau:

Taurine

PEA:

Phosphoethanolamine

AADA:

2-aminoadipic acid

AABA:

α-aminobutyric acid

asn:

Asparagine

cys:

Cystine

orn:

Ornithine

EA:

Ethanolamine

BCAA:

Branched chain amino acids (leu, ile, val)

LNAA:

Large neutral amino acids, including phe, tyr, trp, thr, met, gln, his, val, ile, leu

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Acknowledgments

The authors acknowledge the technical assistance of Ms. Erica Thorson.

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Authors and Affiliations

  1. Division of Experimental and Systems Pharmacology, College of Pharmacy, Washington State University, Pharmaceutical and Biomedical Sciences Building, Room 347, 412 E. Spokane Falls Blvd, Spokane, WA, 99204, USA

    Kara R. Vogel & K. Michael Gibson

  2. Kimberly H. Courtwright and Joseph W. Summers Institute of Metabolic Disease, Baylor Research Institute, Dallas, TX, USA

    Erland Arning & Teodoro Bottiglieri

Authors
  1. Kara R. Vogel
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  2. Erland Arning
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  3. Teodoro Bottiglieri
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  4. K. Michael Gibson
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Corresponding author

Correspondence to K. Michael Gibson.

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Animal rights

All institutional and national guidelines for the care and use of laboratory animals were followed.

Additional information

Communicated by: Nenad Blau

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Supplemental Fig. 1

Concentrations of small neutral amino acids in brain, liver, and sera derived from control and PKU mice receiving 18 %, 6 %, and 4 % protein diets for 3 weeks. Abbreviations employed: gly, glycine; pro, proline; ala, alanine; β−ala, β-alanine; ser, serine. Other parameters are described in the legend for Fig. 1. β-alanine was not detectable in sera. (GIF 205 kb)

High resolution image (TIF 497 kb)

Supplemental Fig. 2

Concentrations of other amino acids, including non-protein amino acids and metabolites, in brain, liver, and sera derived from control and PKU mice receiving 18 %, 6 %, and 4 % protein diets for 3 weeks. Abbreviations employed: cit, citrulline; tau, taurine; PEA, phosphoethanolamine; AADA, 2-aminoadipic acid; AABA, α-aminobutyric acid. AADA represents a metabolite on the lysine/saccharopine catabolic pathway. AABA is also recognized structurally as homoalanine, and is generated via transamination of 2-oxobutyrate, a metabolite in the isoleucine biosynthetic pathway. Other parameters are described in the legend for Fig. 1. Neither AADA nor AABA were detectable in sera. (GIF 182 kb)

High resolution image (TIF 457 kb)

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Vogel, K.R., Arning, E., Bottiglieri, T. et al. Multicompartment analysis of protein-restricted phenylketonuric mice reveals amino acid imbalances in brain. J Inherit Metab Dis 40, 227–235 (2017). https://doi.org/10.1007/s10545-016-9984-3

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  • DOI: https://doi.org/10.1007/s10545-016-9984-3

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