Abstract
The creatine/phosphocreatine system is essential for cellular phosphate coupled energy storage and production. We investigated the utility of creatine monohydrate supplementation in two different creatine deficient knockout mouse models. Following weaning, female Arginine: Glycine Amidinotransferase (AGAT) and Guanidinoacetate: methyltransferase (GAMT) knockouts and wild type mice were studied based on their genotypes and dietary supplementation (creatine free or 2% creatine monohydrate supplemented diet) for 10 weeks, using a series of behavioral tests and biochemical analyzes. An improved Rota rod performance was observed in both AGAT (p = 0.02) and GAMT knockout mice (p < 0.001) supplemented with 2% creatine. During Morris water maze probe trial, creatine supplemented AGAT knockout mice took less time to reach virtual platform (p = 0.03) and more frequently crossed this area (p = 0.001) than mice on creatine free diet. Similar observations were recorded for GAMT knockout mice. Urinary creatinine concentrations for AGAT (p = 0.001) and GAMT (p = 0.05) knockout mice were increased following creatine supplementation. Creatine supplementation has a potential to improve neuro-muscular coordination, spatial learning in both AGAT and GAMT knockout mice. Long term Creatine supplementation results in increased urine creatinine concentrations indicating improved creatine metabolism in knockout mice.
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References
AlmediaL S, Vilarinho L, Darmin PS, Rosenberg EH, Martinez-Munoz C, Jakobs C et al (2007) A prevalent pathogenic GAMT mutation (c.59G>C) in Portugal. Mol Genet Metab 91:1–6
Almeida LS, Salomons GS, Hogenboom F, Jakobs C, Schoffelmeer AN (2006) Exocytotic release of creatine in rat brain. Synapse 60(2):118–123
Amital D, Vishne T, Roitman S, Kotler M, Levine J (2006) Open study of creatinemonohydrate in treatment-resistant posttraumatic stress disorder. J Clin Psychiat 67:836–837
Battini B, Euzzi V, Carducci C, Tosetti M, Binachi MC, Item CB et al (2002) Creatine depletion in a new case with AGAT deficiency: clinical and genetic study in a large pedigree. Mol Gen Metab 77:326–331
Bianchi MC, Tosetti M, Fornai F, Alessandri MG, Cipriani P, De Vito G et al (2000) Reversible brain creatine deficiency in two sisters with normal blood creatine level. Ann Neurol 47:511–513
Bianchi MC, Tosetti M, Battini R, Leuzzi V, Alessandri MG, Carducci C et al (2007) Treatment monitoring of brain creatine deficiency syndromes: a 1H- and 31P-MR spectroscopy study. Am J Neuroradiol 28:548–554
Bodamer OA, Bloesch SM, Gregg AR, Stockler-Ipsiroglu S, Brien EO (2001) Analysis of guanidinoacetate and creatine by isotope dilution electrospray tandem mass spectrometry. Clin Chim Acta 308:173–178
Cecil M, Salomons WS, Ball B, Wong G, Chuck NM, Verhoeven CJ et al (2001) Reversible brain creatine deficiency with elevated serum and urine creatine: a creatine transporter defect? Ann Neurol 49:401–404
Choe C, Nabuurs C, Stockebr M, Neu A, Nune P, Morellini F et al (2013) L-arginine:glycineamidinotransferase (AGAT) deficiency protects from metabolic syndrome. Human Mol Genet 22(1):110–123
Dhar SU, Scaglia F, Li FY, Smith L, Barshop BA, Eng CM et al (2009) Expanded clinical and molecular spectrum of guanidinoacetate methyltransferase (GAMT) deficiency. Mol Genet Metabol 96:38–43
Figura V (2001) Hanefeld F, Isbrandt D. StocklerIpsiroglu S. Guanidinoacetate methyltransferase deficiency. The metabolic and molecular basis of inherited disease. McGraw Hill, New York
Gualano B, Novaes RB, Artioli GG, Freire TO, Coelho DF, Scagliusi FB et al (2007) Effects of creatine supplementation on glucose tolerance and insulin sensitivity in sedentary healthy males undergoing aerobic training. Amino Acids 34:245–250
Humm A, Fritsche E, Steinbacher S, Huber R (1997) Crystal structure and mechanism of human L-arginine:glycineamidinotransferase: a mitochondrial enzyme involved in creatine biosynthesis. EMBO J 16:3373
Iqbal F (2009) Neuroprotective roleof creatine defined in mouse models of Arginine: Glycine Amidino Transferase (AGAT) and Guanidinoacetate Methyltransferase (GAMT) deficiency. PhD Thesis. Medical University Vienna, Austria
Iqbal F (2015) Human guanidinoacetate n-methyl transferase (GAMT) deficiency:a treatable inborn error of metabolism. Pak J Pharmaceut Sci 28(6):2207–2211
Iqbal S, Ali M, Iqbal F (2015a) Long-term creatine monohydrate supplementation, following neonatal hypoxic ischemic insult, improves neuromuscular coordination and spatial learning in male albinomouse. Brain Res 1603:76–83
Iqbal S, Ali M, Akbar A, Iqbal F (2015b) Effectsofdietary Creatine supplementationfor 8 weeks on neuromuscular coordination and learning in male albino mouse following neonatal hypoxic ischemic insult. Neurol Sci 36(5):765–770
Item CB (2004) Mercimek-Mahmutoglu, Battini R, Edlinger-Horvat C, Stromberger C, Bodamer O, et al., characterization of seven novel mutations in seven patients with GAMT deficiency. Hum Mut 23:524–530
Item CB, Stöckler-Ipsiroglu S, Stromberger C, Mühl A, Alessandari MG, Bianchi MC et al (2001) Arginine:glycineamidinotransferase deficiency: the third inborn error of metabolism in man. Am J Hum Genet 69:1127–1133
Komura K, Hobbiebrunken E, Wilichowski EK, Henefeld FA (2003) Effectiveness of creatine monohydrate in mitochondrial encephalomyopathies. Pediatr Neurol 28:53–58
Kreider RB, Ferreira M, Wilson M, Grindstaff P, Plisk S, Reinardy J et al (1998) Effects of creatine supplementation on body composition, strength and sprint performance. Med Sci Sport Exer 30:73–82
Mercimek-Mahmutoglu S, Stoeckler-Ipsiroglu S, Adami A, Appleton R, Araújo HC, Duran M et al (2006) GAMT deficiency: features, treatment, and outcome in an inborn error of creatine synthesis. Neurol 67:480–484
MuddSH EMH, Scriver CR (1980) Labile methyl group balances in the human: the role of sarcosine. Metabol 29:707–720
Nabuurs CI, Choe CU, Veltien A, Kan HE, van Loon LJC, Rodenburg RJT et al (2013) Disturbed energy metabolism and muscular dystrophy caused by pure creatine deficiency are reversible by creatine intake. J Physiol 591(2):571–592
Rae C, Digney AL, Mc Ewan SR, Bates TC (2003) Oral creatine monohydrate supplementation improves brain performance: a double-blind, placebo controlled, cross-over trial. Proc Biol Sci 270:2147–2150
Salmons GS, van Dooren SJM, Verhoeven NM, Cecil KM, Ball WS, DeGrauw TS et al (2001) X-linked creatine transporter (SLC6A8 gene) defect: a new creatine deficiency syndrome. Am J Hum Genet 68:1497–1500
Schmidt A, Marescau B, Boehm EA, Renema WKJ, Peco R, Das A et al (2004) Severely altered guanidino compound levels, disturbed body weight homeostasis and impaired fertility in a mouse model of guanidinoacetate N-methyltransferase (GAMT) deficiency. Hum Mol Genet 13:905–921
Schulze A, Hess T, Wevers R, Mayatepek E, Bachert P, Marescau BV et al (1997) Creatine deficiency syndrome caused by guanidinoacetate methyltransferase deficiency: diagnostic tools or a new inborn error of metabolism. J Pediatr 131:626–631
Sipilä I, Simell O, Arjoma P (1980) Gyrate atrophy of the choroid and retina with hyperornithinemia. J Clin Invest 6:684
Stöckler S, Holzbach U, Hanefeld F, Marquardt I, Helms M, Requart M et al (1994) Creatine deficiency in the brain: a new, treatable inborn error of metabolism. Pediatr Res 36:409–413
Stöckler S, Marescau B, De Deyn PP, Trijbels JMF, Hanefeld F (1997) Guanidino compounds in guanidinoacetate methyltransferase deficiency, a new inborn error of creatine synthesis. Metabol 46:1189–1193
Torremans A, Marescau B, Possemiers I, Dam DV, Hooge RD, Isbrandt D et al (2005) Biochemical and behavioural phenotyping of a mouse model for GAMT deficiency. J Neuol Sci 231:49–55
Valenzuela MJ, Jones M, Wen W, Rae C, Graham S, Shnier R et al (2003) Memory training alters hippocampal neurochemistry in healthy elderly. Neuro report 14:1333–1337
Wallimann T, Tokarska-Schlattner M, Schlattner U (2011) The creatine kinase system and pleiotropic effects of creatine. Ami Acids 40:1271–1296
Watanabe A, Kato N, Kato T (2001) Effects of creatine on mental fatigue and cerebral hemoglobin oxygenation. Neurosci Res 42:279–285
Wyss M, Kaddurah-Daouk R (2000) Creatine and creatinine metabolism. Physiol Rev 80:1107–1213
Acknowledgements
The authors are grateful to Prof. Dr. Dirk Isbrandt, University of Hamburg, Hamburg, Germany for donating breeding pairs of AGAT and GAMT mouse models.
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This work was part of the PhD (FI). He was sponsored by the Higher Education Commission (HEC) of Pakistan through their overseas PhD fellowship program.
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Supplementary Figure 1
Comparison of weight gain in the four experimental treatments for AGAT (A) and GAMT (B) mice after 10 weeks of special diet supplementation. (JPEG 104 kb)
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Supplementary Figure Table 1 (DOC 38.5 kb)
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Supplementary Figure Table 2 (DOC 37.5 kb)
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Iqbal, F., Hoeger, H., Lubec, G. et al. Biochemical and behavioral phenotype of AGAT and GAMT deficient mice following long-term Creatine monohydrate supplementation. Metab Brain Dis 32, 1951–1961 (2017). https://doi.org/10.1007/s11011-017-0092-3
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DOI: https://doi.org/10.1007/s11011-017-0092-3