Abstract
Autism is a severe developmental disorder, whose pathogenetic underpinnings are still largely unknown. Temporocortical gray matter from six matched patient–control pairs was used to perform post-mortem biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier (AGC), which participates in the aspartate/malate reduced nicotinamide adenine dinucleotide shuttle and is physiologically activated by calcium (Ca2+). AGC transport rates were significantly higher in tissue homogenates from all six patients, including those with no history of seizures and with normal electroencephalograms prior to death. This increase was consistently blunted by the Ca2+ chelator ethylene glycol tetraacetic acid; neocortical Ca2+ levels were significantly higher in all six patients; no difference in AGC transport rates was found in isolated mitochondria from patients and controls following removal of the Ca2+-containing postmitochondrial supernatant. Expression of AGC1, the predominant AGC isoform in brain, and cytochrome c oxidase activity were both increased in autistic patients, indicating an activation of mitochondrial metabolism. Furthermore, oxidized mitochondrial proteins were markedly increased in four of the six patients. Variants of the AGC1-encoding SLC25A12 gene were neither correlated with AGC activation nor associated with autism-spectrum disorders in 309 simplex and 17 multiplex families, whereas some unaffected siblings may carry a protective gene variant. Therefore, excessive Ca2+ levels are responsible for boosting AGC activity, mitochondrial metabolism and, to a more variable degree, oxidative stress in autistic brains. AGC and altered Ca2+ homeostasis play a key interactive role in the cascade of signaling events leading to autism: their modulation could provide new preventive and therapeutic strategies.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
American Psychiatric Association. Diagnostic and Statistical Manual of Mental Disorders, 4th edn. American Psychiatric Association: Washington, DC, 1994.
Rutter M . Incidence of autism spectrum disorders: changes over time and their meaning. Acta Paediatr 2005; 94: 2–15.
Persico AM, Bourgeron T . Searching for ways out of the autism maze: genetic, epigenetic and environmental clues. Trends Neurosci 2006; 29: 349–358.
Piven J, Palmer P, Jacobi D, Childress D, Arndt S . Broader autism phenotype: evidence from a family history study of multiple-incidence autism families. Am J Psychiatry 1997; 154: 185–190.
Berney TP . Autism, an evolving concept. Br J Psychiatry 2000; 176: 20–25.
Miller MT, Stromland K, Ventura L, Johansson M, Bandim JM, Gillberg C . Autism associated with conditions characterized by developmental errors in early embryogenesis: a mini review. Int J Dev Neurosci 2005; 23: 201–219.
Bauman ML, Kemper TL . Neuroanatomic observations of the brain in autism: a review and future directions. Int J Dev Neurosci 2005; 23: 183–187.
Teitelbaum O, Benton T, Shah PK, Prince A, Kelly JL, Teitelbaum P . Eshkol-Wachman movement notation in diagnosis: the early detection of Asperger's syndrome. Proc Natl Acad Sci USA 2004; 101: 11909–11914.
Sacco R, Militerni R, Frolli A, Bravaccio C, Gritti A, Elia M et al. Clinical, morphological, and biochemical correlates of head circumference in autism. Biol Psychiatry 2007; 62: 1038–1047.
Wakefield AJ, Ashwood P, Limb K, Anthony A . The significance of ileo-colonic lymphoid nodular hyperplasia in children with autistic spectrum disorder. Eur J Gastroenterol Hepatol 2005; 17: 827–836.
Jyonouchi H, Geng L, Ruby A, Zimmerman-Bier B . Dysregulated innate immune responses in young children with autism spectrum disorders: their relationship to gastrointestinal symptoms and dietary intervention. Neuropsychobiology 2005; 51: 77–85.
Reichelt WH, Knivsberg AM, Nodland M, Stensrud M, Reichelt KL . Urinary peptide levels and patterns in autistic children from seven countries, and the effect of dietary intervention after 4 years. Dev Brain Dysfunct 1997; 10: 44–55.
Chugani DC, Sundram BS, Behen M, Lee ML, Moore GJ . Evidence of altered energy metabolism in autistic children. Prog Neuropsychopharmacol Biol Psychiatry 1999; 23: 635–641.
Correia C, Coutinho AM, Diogo L, Grazina M, Marques C, Miguel T et al. Brief report: high frequency of biochemical markers for mitochondrial dysfunction in autism: no association with the mitochondrial aspartate/glutamate carrier SLC25A12 gene. J Autism Dev Disord 2006; 36: 1137–1140.
Ramoz N, Reichert JG, Smith CJ, Silverman JM, Bespalova IN, Davis KL et al. Linkage and association of the mitochondrial aspartate/glutamate carrier SLC25A12 gene with autism. Am J Psychiatry 2004; 161: 662–669.
Segurado R, Conroy J, Meally E, Fitzgerald M, Gill M, Gallagher L . Confirmation of association between autism and the mitochondrial aspartate/glutamate carrier SLC25A12 gene on chromosome 2q31. Am J Psychiatry 2005; 162: 2182–2184.
Palmieri F . The mitochondrial transporter family (SLC25): physiological and pathological implications. Pflugers Arch 2004; 447: 689–709.
Palmieri L, Pardo B, Lasorsa FM, del Arco A, Kobayashi K, Iijima M et al. Citrin and aralar1 are Ca2+-stimulated aspartate/glutamate transporters in mitochondria. EMBO J 2001; 20: 5060–5069.
Satrustegui J, Pardo B, del Arco A . Mitochondrial transporters as novel targets for intracellular calcium signaling. Physiol Rev 2007; 87: 29–67.
Ramos M, del Arco A, Pardo B, Martinez-Serrano A, Martinez-Morales JR, Kobayashi K et al. Developmental changes in the Ca2+-regulated mitochondrial aspartate–glutamate carrier aralar1 in brain and prominent expression in the spinal cord. Brain Res Dev Brain Res 2003; 143: 33–46.
del Arco A, Morcillo J, Martinez-Morales JR, Galian C, Martos V, Bovolenta P et al. Expression of the aspartate/glutamate mitochondrial carriers aralar1 and citrin during development and in adult rat tissues. Eur J Biochem 2002; 269: 3313–3320.
Tufty RM, Kretsinger RH . Troponin and parvalbumin calcium binding regions predicted in myosin light chain and T4 lysozyme. Science 1975; 187: 167–169.
Lasorsa FM, Pinton P, Palmieri L, Fiermonte G, Rizzuto R, Palmieri F . Recombinant expression of the Ca2+-sensitive aspartate/glutamate carrier increases mitochondrial ATP production in agonist-stimulated CHO cells. J Biol Chem 2003; 278: 38686–38692.
Brookes PS, Yoon Y, Robotham JL, Anders MW, Sheu SS . Calcium, ATP, and ROS: a mitochondrial love–hate triangle. Am J Physiol Cell Physiol 2004; 287: C817–C833.
Zilbovicius M, Meresse I, Chabane N, Brunelle F, Samson Y, Boddaert N . Autism, the superior temporal sulcus and social perception. Trends Neurosci 2006; 29: 359–366.
Campbell DB, D'Oronzio R, Garbett K, Ebert PJ, Mirnics K, Levitt P et al. Disruption of cerebral cortex MET signaling in autism spectrum disorder. Ann Neurol 2007; 62: 243–250.
Lintas C, Sacco R, Garbett K, Mirnics K, Militerni R, Bravaccio C et al. Involvement of the PRKCB1 gene in autistic disorder: significant genetic association and reduced neocortical gene expression. Mol Psychiatry 2008; [e-pub ahead of print].
Garbett K, Ebert PJ, Mitchell A, Lintas C, Manzi B, Mirnics K et al. Immune transcriptome alterations in the temporal cortex of subjects with autism. Neurobiol Dis 2008; 30: 303–311.
Darley-Usmar VM, Rickwood D, Wilson MT (eds). Mitochondria, A Practical Approach. IRL Press: Washington, DC, 1987.
Palmieri F, Indiveri C, Bisaccia F, Iacobazzi V . Mitochondrial metabolite carrier proteins: purification, reconstitution, and transport studies. Methods Enzymol 1995; 260: 349–369.
Grynkiewicz G, Poenie M, Tsien RY . A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 1985; 260: 3440–3450.
Conciatori M, Stodgell CJ, Hyman SL, O'Bara M, Militerni R, Bravaccio C et al. Morphogenetic effect of the HOXA1 A218G polymorphism on head circumference in patients with autism. Biol Psychiatry 2004; 55: 413–419.
Lord C, Rutter M, DiLavore PC, Risi S . ADOS, Autism Diagnostic Observation Schedule. Western Psychological Services: Los Angeles, 2002 (Italian version ed. by Tancredi R, Saccani M, Persico AM, Parrini B, Igliozzi R and Faggioli R. Organizzazioni Speciali: Florence, 2005).
Rutter M, Le Couter A, Lord C . ADI-R, Autism Diagnostic Interview—Revised. Western Psychological Services: Los Angeles, 2003 (Italian version ed. by Faggioli R, Saccani M, Persico AM, Tancredi R, Parrini B and Igliozzi R. Organizzazioni Speciali: Florence, 2005).
Sacco R, Papaleo V, Hager J, Rousseau F, Moessner R, Militerni R et al. Case–control and family-based association studies of candidate genes in autistic disorder and its endophenotypes: TPH2 and GLO1. BMC Med Genet 2007; 8: 11.
Barrett JC, Fry B, Maller J, Daly MJ . Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics 2005; 21: 263–265.
Horvath S, Xu X, Lake SL, Silverman EK, Weiss ST, Laird NM . Family-based tests for associating haplotypes with general phenotype data: application to asthma genetics. Genet Epidemiol 2004; 26: 61–69.
Spielman RS, Ewens WJ . The TDT and other family-based tests for linkage disequilibrium and association. Am J Hum Genet 1996; 59: 983–989.
Dudbridge F . Pedigree disequilibrium tests for multilocus haplotypes. Genet Epidemiol 2003; 25: 115–121.
Gabriel SB, Schaffner SF, Nguyen H, Moore JM, Roy J, Blumenstiel B et al. The structure of haplotype blocks in the human genome. Science 2002; 296: 2225–2229.
Pritchard JK, Stephens M, Donnelly P . Inference of population structure using multilocus genotype data. Genetics 2000; 155: 945–959.
Krey JF, Dolmetsch RE . Molecular mechanisms of autism: a possible role for Ca2+ signaling. Curr Opin Neurobiol 2007; 17: 112–119.
Splawski I, Timothy KW, Sharpe LM, Decher N, Kumar P, Bloise R et al. Ca(V)1.2 calcium channel dysfunction causes a multisystem disorder including arrhythmia and autism. Cell 2004; 119: 19–31.
Hope CI, Sharp DM, Hemara-Wahanui A, Sissingh JI, Lundon P, Mitchell EA et al. Clinical manifestations of a unique X-linked retinal disorder in a large New Zealand family with a novel mutation in CACNA1F, the gene responsible for CSNB2. Clin Experiment Ophthalmol 2005; 33: 129–136.
Laumonnier F, Roger S, Guerin P, Molinari F, M'rad R, Cahard D et al. Association of a functional deficit of the BKCa channel, a synaptic regulator of neuronal excitability, with autism and mental retardation. Am J Psychiatry 2006; 163: 1622–1629.
Rizzuto R, Bernardi P, Pozzan T . Mitochondria as all-round players of the calcium game. J Physiol 2000; 529: 37–47.
Vargas DL, Nascimbene C, Krishnan C, Zimmerman AW, Pardo CA . Neuroglial activation and neuroinflammation in the brain of patients with autism. Ann Neurol 2005; 57: 67–81.
Jin D, Liu HX, Hirai H, Torashima T, Nagai T, Lopatina O et al. CD38 is critical for social behaviour by regulating oxytocin secretion. Nature 2007; 446: 41–45.
Kumar RA, KaraMohamed S, Sudi J, Conrad DF, Brune C, Badner JA et al. Recurrent 16p11.2 microdeletions in autism. Hum Mol Genet 2008; 17: 628–638.
Lohiya GS, Tan-Figueroa L, Iannucci A . Identification of low bone mass in a developmental center: finger bone mineral density measurement in 562 residents. J Am Med Dir Assoc 2004; 5: 371–376.
Jaffe JS, Timell AM, Gulanski BI . Prevalence of low bone density in women with developmental disabilities. J Clin Densitom 2001; 4: 25–29.
Blasi F, Bacchelli E, Carone S, Toma C, Monaco AP, Bailey AJ et al. SLC25A12 and CMYA3 gene variants are not associated with autism in the IMGSAC multiplex family sample. Eur J Hum Genet 2006; 14: 123–126.
Rabionet R, McCauley JL, Jaworski JM, Ashley-Koch AE, Martin ER, Sutcliffe JS et al. Lack of association between autism and SLC25A12. Am J Psychiatry 2006; 163: 929–931.
Silverman JM, Buxbaum JD, Ramoz N, Schmeidler J, Reichenberg A, Hollander E et al. Autism-related routines and rituals associated with a mitochondrial aspartate/glutamate carrier SLC25A12 polymorphism. Am J Med Genet B Neuropsychiatr Genet 2007; 147B: 408–410.
Chauhan A, Chauhan V . Oxidative stress in autism. Pathophysiology 2006; 13: 171–181.
Lepagnol-Bestel AM, Maussion G, Boda B, Cardona A, Iwayama Y, Delezoide AL et al. SLC25A12 expression is associated with neurite outgrowth and is upregulated in the prefrontal cortex of autistic subjects. Mol Psychiatry 2008; 13: 385–397.
Jacquemont ML, Sanlaville D, Redon R, Raoul O, Cormier-Daire V, Lyonnet S et al. Array-based comparative genomic hybridisation identifies high frequency of cryptic chromosomal rearrangements in patients with syndromic autism spectrum disorders. J Med Genet 2006; 43: 843–849.
Autism Genome Project Consortium, Szatmari P, Paterson AD, Zwaigenbaum L, Roberts W, Brian J, Liu XQ et al. Mapping autism risk loci using genetic linkage and chromosomal rearrangements. Nat Genet 2007; 39: 319–328.
Bernard S, Enayati A, Roger H, Binstock T, Redwood L . The role of mercury in the pathogenesis of autism. Mol Psychiatry 2002; 7: S42–S43.
Nelson KB, Bauman ML . Thimerosal and autism? Pediatrics 2003; 111: 674–679.
Elferink JG . Thimerosal: a versatile sulfhydryl reagent, calcium mobilizer, and cell function-modulating agent. Gen Pharmacol 1999; 33: 1–6.
Burbacher TM, Shen DD, Liberato N, Grant KS, Cernichiari E, Clarkson T . Comparison of blood and brain mercury levels in infant monkeys exposed to methylmercury or vaccines containing thimerosal. Environ Health Perspect 2005; 113: 1015–1021.
Hornig M, Chian D, Lipkin WI . Neurotoxic effects of postnatal thimerosal are mouse strain dependent. Mol Psychiatry 2004; 9: 833–845.
Brown MJ, Willis T, Omalu B, Leiker R . Deaths resulting from hypocalcemia after administration of edetate disodium: 2003–2005. Pediatrics 2006; 118: e534–e536.
Sinha Y, Silove N, Williams K . Chelation therapy and autism. BMJ 333: 756.
Acknowledgements
We gratefully acknowledge all the patients and families who generously contributed to these studies, the Autism Tissue Program, Harvard Brain Tissue Resource Center and NICHD Brain and Tissue Bank for providing the brain tissue samples, and Roberto Rigardetto and Franco Nardocci for contributing to patient recruitment. This work was supported by MIUR PRIN 2005052128 and 2006058195 (LP, PS and AMP), MIUR FIRB (LP) and the Fondation Jerome Lejeune (AMP).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Supplementary Information accompanies the paper on the Molecular Psychiatry website (http://www.nature.com/mp)
Rights and permissions
About this article
Cite this article
Palmieri, L., Papaleo, V., Porcelli, V. et al. Altered calcium homeostasis in autism-spectrum disorders: evidence from biochemical and genetic studies of the mitochondrial aspartate/glutamate carrier AGC1. Mol Psychiatry 15, 38–52 (2010). https://doi.org/10.1038/mp.2008.63
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/mp.2008.63
Keywords
This article is cited by
-
Repurposing Niclosamide as a plausible neurotherapeutic in autism spectrum disorders, targeting mitochondrial dysfunction: a strong hypothesis
Metabolic Brain Disease (2023)
-
Calciopathies and Neuropsychiatric Disorders: Physiological and Genetic Aspects
Neuroscience and Behavioral Physiology (2023)
-
The potential involvement of inhaled iron (Fe) in the neurotoxic effects of ultrafine particulate matter air pollution exposure on brain development in mice
Particle and Fibre Toxicology (2022)
-
Assessment of Urinary Lead (Pb) and Essential Trace Elements in Autism Spectrum Disorder: a Case-Control Study Among Preschool Children in Malaysia
Biological Trace Element Research (2022)
-
R.ROSETTA: an interpretable machine learning framework
BMC Bioinformatics (2021)