Creatine transport and pathological changes in creatine transporter deficient mice
Adam M. Wawro
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorChandresh R. Gajera
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorSteven A. Baker
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorJeffrey J. Nirschl
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorHannes Vogel
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorCorresponding Author
Thomas J. Montine
Department of Pathology, Stanford University, Stanford, California, USA
Correspondence
Thomas J. Montine, Department of Pathology, School of Medicine, Stanford University, 300 Pasteur Dr Rm 235, Stanford, CA 94305, USA.
Email: [email protected]
Search for more papers by this authorAdam M. Wawro
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorChandresh R. Gajera
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorSteven A. Baker
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorJeffrey J. Nirschl
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorHannes Vogel
Department of Pathology, Stanford University, Stanford, California, USA
Search for more papers by this authorCorresponding Author
Thomas J. Montine
Department of Pathology, Stanford University, Stanford, California, USA
Correspondence
Thomas J. Montine, Department of Pathology, School of Medicine, Stanford University, 300 Pasteur Dr Rm 235, Stanford, CA 94305, USA.
Email: [email protected]
Search for more papers by this authorFunding information: Farmer Family Foundation; Stanford ChEM-H Metabolomics Knowledge Center
Abstract
The severe impact on brain function and lack of effective therapy for patients with creatine (Cr) transporter deficiency motivated the generation of three ubiquitous Slc6a8 deficient mice (−/y). While each mouse knock-out line has similar behavioral effects at 2 to 3 months of age, other features critical to the efficient use of these mice in drug discovery are unclear or lacking: the concentration of Cr in brain and heart differ widely between mouse lines, there are limited data on histopathologic changes, and no data on Cr uptake. Here, we determined survival, measured endogenous Cr and uptake of its deuterium-labeled analogue Cr-d3 using a liquid chromatography coupled with tandem mass spectrometry assay, and performed comprehensive histopathologic examination on the Slc6a8−/y mouse developed by Skelton et al. Our results show that Slc6a8−/y mice have widely varying organ-specific uptake of Cr-d3, significantly diminished growth with the exception of brain, progressive vacuolar myopathy, and markedly shortened lifespan.
CONFLICT OF INTEREST
The authors declare that they have no conflict of interest.
Open Research
DATA AVAILABILITY STATEMENT
Data generated during this study are available from the corresponding author on reasonable request.
Supporting Information
| Filename | Description |
|---|---|
| jimd12358-sup-0001-Figures.docxWord 2007 document , 17.6 MB | Figure S1 Formalin-fixed paraffin-embedded sections of muscle from WT and Slc6a8−/y mice aged 6 months and stained H&E, PAS with and without diastase pre-treatment shows no evidence of PAS-positive or PAS-D resistant inclusions (magnification main = 200×, inset = 400×. Scale bar = 50 μm). Figure S2. Formalin-fixed paraffin-embedded sagittal sections of the brain including representative cortex, hippocampus, and cerebellum at 6 months show no significant histopathologic changes in wild type and knockout tissue on expert evaluation (magnification = 100×. Scale bar = 100 μm). Note that neuroanatomical sections were not stereotactically obtained and do not necessarily represent identical sagittal planes between two conditions. Figure S3. Formalin-fixed paraffin-embedded sections of the kidney, liver, testes, and cardiac muscle (ventricle) at 6 months show no significant histopathologic changes in wild type and knockout tissue on expert evaluation (magnification = 100×. Scale bar = 100 μm). |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Cantoni GL, Vignos PJ. Enzymatic mechanism of creatine synthesis. J Biol Chem. 1954; 209(2): 647-659.
- 2Wyss M, Kaddurah-Daouk R. Creatine and creatinine metabolism. Physiol Rev. 2000; 80(3): 1107-1213.
- 3Ohtsuki S, Tachikawa M, Takanaga H, et al. The blood–brain barrier creatine transporter is a major pathway for supplying creatine to the brain. J Cereb Blood Flow Metab. 2002; 22(11): 1327-1335.
- 4Braissant O, Henry H, Loup M, Eilers B, Bachmann C. Endogenous synthesis and transport of creatine in the rat brain: an in situ hybridization study. Mol Brain Res. 2001; 86(1–2): 193-201.
- 5Perasso L, Cupello A, Lunardi GL, Principato C, Gandolfo C, Balestrino M. Kinetics of creatine in blood and brain after intraperitoneal injection in the rat. Brain Res. 2003; 974(1): 37-42.
- 6Salomons GS, van Dooren SJ, Verhoeven NM, et al. X-linked creatine-transporter gene (SLC6A8) defect: a new creatine-deficiency syndrome. Am J Hum Genet. 2001; 68(6): 1497-1500.
- 7Cecil KM, Salomons GS, Ball WS Jr, et al. Irreversible brain creatine deficiency with elevated serum and urine creatine: a creatine transporter defect? Ann Neurol. 2001; 49(3): 401-404.
- 8de Grauw TJ, Salomons GS, Cecil KM, et al. Congenital creatine transporter deficiency. Neuropediatrics. 2002; 33(5): 232-238.
- 9Salomons GS, Van Dooren SJM, Verhoeven NM, et al. X-linked creatine transporter defect: an overview. J Inherit Metab Dis. 2003; 26(2–3): 309-318.
- 10de Grauw TJ, Cecil KM, Byars AW, Salomons GS, Ball WS, Jakobs C. The clinical syndrome of creatine transporter deficiency. Mol Cell Biochem. 2003; 244(1–2): 45-48.
- 11Anselm IA, Alkuraya FS, Salomons GS, et al. X-linked creatine transporter defect: a report on two unrelated boys with a severe clinical phenotype. J Inherit Metab Dis. 2006; 29(1): 214-219.
- 12Póo-Argüelles P, Arias A, Vilaseca MA, et al. X-linked creatine transporter deficiency in two patients with severe mental retardation and autism. J Inherit Metab Dis. 2006; 29(1): 220-223.
- 13Mancardi MM, Caruso U, Schiaffino MC, et al. Severe epilepsy in X-linked creatine transporter defect (CRTR-D). Epilepsia. 2007; 48(6): 1211-1213. https://doi.org/10.1111/j.1528-1167.2007.01148.x.
- 14Sempere A, Fons C, Arias A, et al. Creatine transporter deficiency in two adult patients with static encephalopathy. J Inherit Metab Dis. 2009; 32(suppl 1): S91-S96.
- 15van de Kamp JM, Betsalel OT, Mercimek-Mahmutoglu S, et al. Phenotype and genotype in 101 males with X-linked creatine transporter deficiency. J Med Genet. 2013; 50(7): 463-472.
- 16Lion-François L, Cheillan D, Pitelet G, et al. High frequency of creatine deficiency syndromes in patients with unexplained mental retardation. Neurology. 2006; 67(9): 1713-1714.
- 17Puusepp H, Kall K, Salomons GS, et al. The screening of SLC6A8 deficiency among Estonian families with X-linked mental retardation. J Inherit Metab Dis. 2010; 33(S3): 5-11. https://doi.org/10.1007/s10545-008-1063-y.
- 18Clark AJ, Rosenberg EH, Almeida LS, et al. X-linked creatine transporter (SLC6A8) mutations in about 1% of males with mental retardation of unknown etiology. Hum Genet. 2006; 119(6): 604-610.
- 19Fons C, Sempere A, Arias A, et al. Arginine supplementation in four patients with X-linked creatine transporter defect. J Inherit Metab Dis. 2008; 31(6): 724-728.
- 20van de Kamp JM, Mancini GM, Salomons GS. X-linked creatine transporter deficiency: clinical aspects and pathophysiology. J Inherit Metab Dis. 2014; 37(5): 715-733.
- 21Bruun TUJ, Sidky S, Bandeira AO, et al. Treatment outcome of creatine transporter deficiency: international retrospective cohort study. Metab Brain Dis. 2018; 33(3): 875-884.
- 22Skelton MR, Schaefer TL, Graham DL, et al. Creatine transporter (CrT; Slc6a8) knockout mice as a model of human CrT deficiency. PLoS One. 2011; 6(1):e16187.
- 23Baroncelli L, Alessandrì MG, Tola J, et al. A novel mouse model of creatine transporter deficiency. F1000Res. 2014; 3: 228.
- 24Stockebrand M, Sasani A, Das D, et al. A mouse model of creatine transporter deficiency reveals impaired motor function and muscle energy metabolism. Front Physiol. 2018; 9: 773.
- 25Kurosawa Y, Degrauw TJ, Lindquist DM, et al. Cyclocreatine treatment improves cognition in mice with creatine transporter deficiency. J Clin Invest. 2012; 122(8): 2837-2846.
- 26Brubaker WF. Creatine prodrugs, compositions and methods of use thereof. US Patent. Published online April 11, 2017. Accessed October 22, 2020. https://patentimages.storage.googleapis.com/58/a5/b8/8d7cdee363a713/US9617230.pdf
- 27Ullio-Gamboa G, Udobi KC, Dezard S, et al. Dodecyl creatine ester-loaded nanoemulsion as a promising therapy for creatine transporter deficiency. Nanomedicine. 2019; 14(12): 1579-1593.
- 28Cacciante F, Gennaro M, Sagona G, et al. Cyclocreatine treatment ameliorates the cognitive, autistic and epileptic phenotype in a mouse model of creatine transporter deficiency. Sci Rep. 2020; 10(1): 18361.
- 29Baroncelli L, Molinaro A, Cacciante F, et al. A mouse model for creatine transporter deficiency reveals early onset cognitive impairment and neuropathology associated with brain aging. Hum Mol Genet. 2016; 25(19): 4186-4200.
- 30Marescau B, Deshmukh DR, Kockx M, et al. Guanidino compounds in serum, urine, liver, kidney, and brain of man and some ureotelic animals. Metabolism. 1992; 41(5): 526-532.
- 31Girardi E, César-Razquin A, Lindinger S, et al. A widespread role for SLC transmembrane transporters in resistance to cytotoxic drugs. Nat Chem Biol. 2020; 16(4): 469-478.
- 32Hanna-El-Daher L, Braissant O. Creatine synthesis and exchanges between brain cells: what can be learned from human creatine deficiencies and various experimental models? Amino Acids. 2016; 48(8): 1877-1895.
- 33Udobi KC, Kokenge AN, Hautman ER, et al. Cognitive deficits and increases in creatine precursors in a brain-specific knockout of the creatine transporter gene Slc6a8. Genes Brain Behav. 2018; 17(6):e12461.
- 34Iyer GS, Krahe R, Goodwin LA, et al. Identification of a testis-expressed creatine transporter gene at 16p11.2 and confirmation of the X-linked locus to Xq28. Genomics. 1996; 34(1): 143-146.
- 35Abplanalp J, Laczko E, Philp NJ, et al. The cataract and glucosuria associated monocarboxylate transporter MCT12 is a new creatine transporter. Hum Mol Genet. 2013; 22(16): 3218-3226.
- 36Joncquel-Chevalier Curt M, Voicu P-M, Fontaine M, et al. Creatine biosynthesis and transport in health and disease. Biochimie. 2015; 119: 146-165.
- 37Farr CV, El-Kasaby A, Freissmuth M, Sucic S. The creatine transporter unfolded: a knotty premise in the cerebral creatine deficiency syndrome. Front Synapt Neurosci. 2020; 12:588954.
- 38Flanagan ME, Cholerton B, Latimer CS, et al. TDP-43 neuropathologic associations in the Nun Study and the Honolulu-Asia Aging Study. J Alzheimers Dis. 2018; 66(4): 1549-1558.
- 39Bharadwaj R, Cimino PJ, Flanagan ME, et al. Application of the condensed protocol for the NIA-AA guidelines for the neuropathological assessment of Alzheimer's disease in an academic clinical practice. Histopathology. 2018; 72(3): 433-440.
- 40Berg S, Kutra D, Kroeger T, et al. ilastik: interactive machine learning for (bio)image analysis. Nat Methods. 2019; 16(12): 1226-1232.
- 41McQuin C, Goodman A, Chernyshev V, et al. CellProfiler 3.0: next-generation image processing for biology. PLoS Biol. 2018; 16(7):e2005970.
