Deletion of the Creatine Transporter (Slc6a8) in Dopaminergic Neurons Leads to Hyperactivity in Mice

  • Zuhair I. Abdulla
  • Bahar Pahlevani
  • Kerstin H. Lundgren
  • Jordan L. Pennington
  • Kenea C. Udobi
  • Kim B. Seroogy
  • Matthew R. SkeltonEmail author


The lack of cerebral creatine (Cr) causes intellectual disability and epilepsy. In addition, a significant portion of individuals with Cr transporter (Crt) deficiency (CTD), the leading cause of cerebral Cr deficiency syndromes (CCDS), are diagnosed with attention-deficit hyperactivity disorder. While the neurological effects of CTD are clear, the mechanisms that underlie these deficits are unknown. Part of this is due to the heterogenous nature of the brain and the unique metabolic demands of specific neuronal systems. Of particular interest related to Cr physiology are dopaminergic neurons, as many CCDS patients have ADHD and Cr has been implicated in dopamine-associated neurodegenerative disorders, such as Parkinson’s and Huntington’s diseases. The purpose of this study was to examine the effect of a loss of the Slc6a8 (Crt) gene in dopamine transporter (Slc6a3; DAT) expressing cells on locomotor activity and motor function as the mice age. Floxed Slc6a8 (Slc6a8flox) mice were mated to DATIREScre expressing mice to generate DAT-specific Slc6a8 knockouts (dCrt−/y). Locomotor activity, spontaneous activity, and performance in the challenging beam test were evaluated monthly in dCrt−/y and control (Slc6a8flox) mice from 3 to 12 months of age. dCrt−/y mice were hyperactive compared with controls throughout testing. In addition, dCrt−/y mice showed increased rearing and hindlimb steps in the spontaneous activity test. Latency to cross the narrow bridge was increased in dCrt−/y mice while foot slips were unchanged. Taken together, these data suggest that the lack of Cr in dopaminergic neurons causes hyperactivity while sparing motor function.


Creatine Creatine transporter DAT-Cre Hyperactivity Motor function 



The authors would like to thank Marla K. Perna and Keila N. Miles for providing scientific input and proofreading this article.

Author Contributions

Mr. Abdulla, Dr. Seroogy, and Dr. Skelton designed the experiment and drafted and edited the manuscript. Mr. Abdulla, Ms. Pahlevani, Ms. Lundgren, Ms. Pennington, and Mr. Udobi conducted and scored all experiments.

Funding Information

This work was supported by National Institutes of Health grant HD080910 and a CARE grant from the Association of Creatine Deficiencies. Portions were supported by the Kerman Family Fund, the Selma Schottenstein Harris Lab for Research in Parkinson’s, the Gardner Family Center for Parkinson’s Disease and Movement Disorders, and the Parkinson’s Disease Support Network, Ohio, Kentucky, and Indiana.


  1. Abdulla, Z. I., J. L. Pennington, A. Gutierrez and M. R. Skelton (2019). “Creatine transporter knockout mice (Slc6a8) show increases in serotonin-related proteins and are resilient to learned helplessness.” bioRxiv: 641845Google Scholar
  2. Backman CM, Malik N, Zhang Y, Shan L, Grinberg A, Hoffer BJ, Westphal H, Tomac AC (2006) Characterization of a mouse strain expressing Cre recombinase from the 3′ untranslated region of the dopamine transporter locus. Genesis 44(8):383–390CrossRefGoogle Scholar
  3. Beninger RJ (1983) The role of dopamine in locomotor activity and learning. Brain Res Rev 6:173–196CrossRefGoogle Scholar
  4. Benjamini Y, Krieger AM, Yekutieli D (2006) Adaptive linear step-up procedures that control the false discovery rate. Biometrika 93(3):491–507CrossRefGoogle Scholar
  5. Birgner C, Nordenankar K, Lundblad M, Mendez JA, Smith C, le Greves M, Galter D, Olson L, Fredriksson A, Trudeau LE, Kullander K, Wallen-Mackenzie A (2010) VGLUT2 in dopamine neurons is required for psychostimulant-induced behavioral activation. Proc Natl Acad Sci U S A 107(1):389–394CrossRefGoogle Scholar
  6. Braissant O, Henry H, Loup M, Eilers B, Bachmann C (2001) Endogenous synthesis and transport of creatine in the rat brain: an in situ hybridization study. Mol Brain Res 86(1–2):193–201CrossRefGoogle Scholar
  7. Cecil KM, Salomons GS, William J, Ball S, Wong B, Chuck G, Verhoeven NM, Jakobs C, DeGrauw TJ (2001) Irreversible brain creatine deficiency with elevated serum and urine creatine: a creatine transporter defect? Ann Neurol 49:401–404CrossRefGoogle Scholar
  8. Cunha MP, Martin-de-Saavedra MD, Romero A, Parada E, Egea J, Del Barrio L, Rodrigues AL, Lopez MG (2013) Protective effect of creatine against 6-hydroxydopamine-induced cell death in human neuroblastoma SH-SY5Y cells: involvement of intracellular signaling pathways. Neuroscience 238:185–194CrossRefGoogle Scholar
  9. Cunha MP, Martin-de-Saavedra MD, Romero A, Egea J, Ludka FK, Tasca CI, Farina M, Rodrigues AL, Lopez MG (2014) Both creatine and its product phosphocreatine reduce oxidative stress and afford neuroprotection in an in vitro Parkinson’s model. ASN Neuro 6(6):175909141455494CrossRefGoogle Scholar
  10. deGrauw TJ, Salomons GS, Cecil KM, Chuck G, Newmeyer A, Schapiro MB, Jakobs C (2002) Congenital creatine transporter deficiency. Neuropediatrics 33:232–238CrossRefGoogle Scholar
  11. Faraone SV (2018) The pharmacology of amphetamine and methylphenidate: relevance to the neurobiology of attention-deficit/hyperactivity disorder and other psychiatric comorbidities. Neurosci Biobehav Rev 87:255–270CrossRefGoogle Scholar
  12. Fleming SM, (2004) Early and Progressive Sensorimotor Anomalies in Mice Overexpressing Wild-Type Human -Synuclein. J. Neurosci 24 (42):9434-9440Google Scholar
  13. Glajch KE, Fleming SM, Surmeier DJ, Osten P (2012) Sensorimotor assessment of the unilateral 6-hydroxydopamine mouse model of Parkinson’s disease. Behav Brain Res 230(2):309–316CrossRefGoogle Scholar
  14. Gomez-Lazaro M, Galindo MF, Concannon CG, Segura MF, Fernandez-Gomez FJ, Llecha N, Comella JX, Prehn JH, Jordan J (2008) 6-Hydroxydopamine activates the mitochondrial apoptosis pathway through p38 MAPK-mediated, p53-independent activation of Bax and PUMA. J Neurochem 104(6):1599–1612CrossRefGoogle Scholar
  15. Hautman ER, Kokenge AN, Udobi KC, Williams MT, Vorhees CV, Skelton MR (2014) Female mice heterozygous for creatine transporter deficiency show moderate cognitive deficits. J Inherit Metab Dis 37(1):63–68CrossRefGoogle Scholar
  16. Hemmerle AM, Dickerson JW, Herring NR, Schaefer TL, Vorhees CV, Williams MT, Seroogy KB (2012) (+/−)3,4-methylenedioxymethamphetamine (“ecstasy”) treatment modulates expression of neurotrophins and their receptors in multiple regions of adult rat brain. J Comp Neurol 520(11):2459–2474CrossRefGoogle Scholar
  17. Hosamani, R., S. R. Ramesh and Muralidhara (2010). “Attenuation of rotenone-induced mitochondrial oxidative damage and neurotoxicty in Drosophila melanogaster supplemented with creatine.” Neurochem Res 35(9): 1402–1412Google Scholar
  18. Klivenyi P, Calingasan NY, Starkov A, Stavrovskaya IG, Kristal BS, Yang L, Wieringa B, Beal MF (2004) Neuroprotective mechanisms of creatine occur in the absence of mitochondrial creatine kinase. Neurobiol Dis 15(3):610–617CrossRefGoogle Scholar
  19. Koob GF, Riley SJ, Smith SC, Robbins TW (1978) Effects of 6-hydroxydopamine lesions of the nucleus accumbens septi and olfactory tubercle on feeding, locomotor activity, and amphetamine anorexia in the rat. J Comp Physiol Psychol 92(5):917–927CrossRefGoogle Scholar
  20. Kostrzewa, J. P., Kostrzewa, R. A., Kostrzewa, R. M., Brus, R., & Nowak, P. (2015). Perinatal 6-Hydroxydopamine Modeling of ADHD. Curr Top Behav Neurosci. doi:10.1007/7854_2015_397Google Scholar
  21. Lowe MT, Kim EH, Faull RL, Christie DL, Waldvogel HJ (2013) Dissociated expression of mitochondrial and cytosolic creatine kinases in the human brain: a new perspective on the role of creatine in brain energy metabolism. J Cereb Blood Flow Metab 33(8):1295–1306CrossRefGoogle Scholar
  22. Matthews RT, Ferrante RJ, Klivenyi P, Yang L, Klein AM, Meuller G, Kaddurah-Daouk R, Beal MF (1999) Creatine and cyclocreatine attenuate MPTP neurotoxicity. Exp Neurol 157:142–149CrossRefGoogle Scholar
  23. Newmeyer A, Cecil KM, Schapiro M, Clark JF, Degrauw TJ (2005) Incidence of brain creatine transporter deficiency in males with developmental delay referred for brain magnetic resonance imaging. Dev Behav Pediatr 26(4):276–282CrossRefGoogle Scholar
  24. Numan S, Gall CM, Seroogy KB (2005) Developmental expression of neurotrophins and their receptors in postnatal rat ventral midbrain. J Mol Neurosci 27(2):245–260CrossRefGoogle Scholar
  25. O’neil B, Gu HH (2013) Amphetamine-induced locomotion in a hyperdopaminergic ADHD mouse model depends on genetic background. Pharmacol Biochem Behav 103 (3):455–459Google Scholar
  26. Ohtsuki S, Tachikawa M, Takanaga H, Shimizu H, Watanabe M, Hosoya K, Terasaki T (2002) The blood-brain barrier creatine transporter is a major pathway for supplying creatine to the brain. J Cereb Blood Flow Metab 22(11):1327–1335CrossRefGoogle Scholar
  27. Palmiter RD (2007) Is dopamine a physiologically relevant mediator of feeding behavior? Trends Neurosci 30(8):375–381CrossRefGoogle Scholar
  28. Pinto M, Nissanka N, Peralta S, Brambilla R, Diaz F, Moraes CT (2016) Pioglitazone ameliorates the phenotype of a novel Parkinson’s disease mouse model by reducing neuroinflammation. Mol. Neurodegener 11 (1).
  29. Przedborski S, Tieu K, Perier C, Vila M (2004) MPTP as a mitochondrial neurotoxic model of Parkinson’s disease. J Bioenerg Biomembr 36(4):375–379CrossRefGoogle Scholar
  30. Roberts DCS, Zis AP, Fibiger HC (1975) Ascending catecholamine pathways and amphetamine-induced locomotor activity: importance of dopamine and apparent non-involvment of norepinephrine. Brain Res 93:441–454CrossRefGoogle Scholar
  31. Rosenberg EH, Almeida LS, Kleefstra T, deGrauw RS, Yntema HG, Bahi N, Moraine C, Ropers H, Fryns J, Degrauw TJ, Jakobs C, Salomons GS (2004) High prevalence of SLC6A8 deficiency in X-linked mental retardation. Am J Hum Genet 75:97–105CrossRefGoogle Scholar
  32. Runegaard AH, Sorensen AT, Fitzpatrick CM, Jorgensen SH, Petersen AV, Hansen NW, Weikop P, Andreasen JT, Mikkelsen JD, Perrier JF, Woldbye D, Rickhag M, Wortwein G, Gether U (2018) Locomotor- and reward-enhancing effects of cocaine are differentially regulated by chemogenetic stimulation of Gi-signaling in dopaminergic neurons. eNeuro 5(3):ENEURO.0345–ENEU17.2018CrossRefGoogle Scholar
  33. Sakai K, Gash DM (1994) Effect of bilateral 6-OHDA lesions of the substantia nigra on locomotor activity in the rat. Brain Res 633:144–150CrossRefGoogle Scholar
  34. Schallert T, Fleming SM, Leasure JL, Tillerson JL, Bland ST (2000) CNS plasticity and assessment of forelimb sensorimotor outcome in unilateral rat models of stroke, cortical ablation, parkinsonism and spinal cord injury. Neuropharmacology 39 (5):777–787Google Scholar
  35. Seroogy KB, Herman JP (1997) In situ hybridization approaches to the study of the nervous system. In: Turner AJ, Bachelard HS (eds) Neurochemistry: a practical approach (2nd edition). Oxford University Press, Oxford, pp 121–150Google Scholar
  36. Skelton MR, Schaefer TL, Graham DL, Degrauw TJ, Clark JF, Williams MT, Vorhees CV (2011) Creatine transporter (CrT; Slc6a8) knockout mice as a model of human CrT deficiency. PLoS One 6(1):e16187CrossRefGoogle Scholar
  37. Tirmenstein MA, Hu CX, Scicchitano MS, Narayanan PK, McFarland DC, Thomas HC, Schwartz LW (2005) Effects of 6-hydroxydopamine on mitochondrial function and glutathione status in SH-SY5Y human neuroblastoma cells. Toxicol in Vitro 19(4):471–479CrossRefGoogle Scholar
  38. Udobi KC, Kokenge AN, Hautman ER, Ullio G, Coene J, Williams MT, Vorhees CV, Mabondzo A, Skelton MR (2018) Cognitive deficits and increases in creatine precursors in a brain-specific knockout of the creatine transporter gene Slc6a8. Genes Brain Behav 17(6):e12461CrossRefGoogle Scholar
  39. Udobi, K. C., N. Delcimmuto, A. N. Kokenge, Z. I. Abdulla, M. K. Perna and M. R. Skelton (2019). “Deletion of the creatine transporter gene in neonatal, but not adult, mice lead to cognitive deficits.” bioRxiv: 582320Google Scholar
  40. Ullio-Gamboa G, Udobi KC, Dezard S, Perna MK, Miles KN, Costa N, Taran F, Pruvost A, Benoit JP, Skelton MR, Lonlay P, Mabondzo A (2019) Dodecyl creatine ester-loaded nanoemulsion as a promising therapy for creatine transporter deficiency. Nanomedicine (London) 14(12):1579–1593CrossRefGoogle Scholar
  41. van de Kamp JM, Betsalel OT, Mercimek-Mahmutoglu S, Abulhoul L, Grunewald S, Anselm I, Azzouz H, Bratkovic D, de Brouwer A, Hamel B, Kleefstra T, Yntema H, Campistol J, Vilaseca MA, Cheillan D, D'Hooghe M, Diogo L, Garcia P, Valongo C, Fonseca M, Frints S, Wilcken B, von der Haar S, Meijers-Heijboer HE, Hofstede F, Johnson D, Kant SG, Lion-Francois L, Pitelet G, Longo N, Maat-Kievit JA, Monteiro JP, Munnich A, Muntau AC, Nassogne MC, Osaka H, Ounap K, Pinard JM, Quijano-Roy S, Poggenburg I, Poplawski N, Abdul-Rahman O, Ribes A, Arias A, Yaplito-Lee J, Schulze A, Schwartz CE, Schwenger S, Soares G, Sznajer Y, Valayannopoulos V, Van Esch H, Waltz S, Wamelink MM, Pouwels PJ, Errami A, van der Knaap MS, Jakobs C, Mancini GM, Salomons GS (2013) Phenotype and genotype in 101 males with X-linked creatine transporter deficiency. J Med Genet 50(7):463–472CrossRefGoogle Scholar
  42. van de Kamp JM, Mancini GM, Salomons GS (2014) X-linked creatine transporter deficiency: clinical aspects and pathophysiology. J Inherit Metab Dis 37(5):715–733CrossRefGoogle Scholar
  43. Wallimann T, Tokarska-Schlattner M, Schlattner U (2011) The creatine kinase system and pleiotropic effects of creatine. Amino Acids 40(5):1271–1296CrossRefGoogle Scholar
  44. Wyss M, Kaddurah-Daouk R (2000) Creatine and creatinine metabolism. Physiol Rev 3(80):1107–1213CrossRefGoogle Scholar
  45. Xiong N, Long X, Xiong J, Jia M, Chen C, Huang J, Ghoorah D, Kong X, Lin Z, Wang T (2012) Mitochondrial complex I inhibitor rotenone-induced toxicity and its potential mechanisms in Parkinson’s disease models. Crit Rev Toxicol 42(7):613–632Google Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of PediatricsUniversity of Cincinnati College of Medicine and Division of Neurology, Cincinnati Children’s Research FoundationCincinnatiUSA
  2. 2.Department of Neurology, College of MedicineUniversity of CincinnatiCincinnatiUSA
  3. 3.Neuroscience Graduate ProgramUniversity of CincinnatiCincinnatiUSA

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