Abstract
Nijmegen breakage syndrome (NBS) is a genomic instability disorder caused by hypomorphic mutations in the Nbs1 gene. When Nbs1 is conditionally inactivated in the central nervous system of mice (Nbs1-CNS-Δ), they suffer from severe cerebellar atrophy, ataxia, and white matter damage. Here, we show that conditional inactivation of the murine Nbs1 gene has a profound effect on the integrity and the functionality of the glial cells, which suggests their crucial role in the pathogenesis of NBS. Interestingly, in Nbs1-CNS-Δ mice, the dramatic reduction in the numbers of Purkinje and granule cells was also linked to a reduction of microglial cells but not to astrocytes (GFAP+), suggesting an impairment in astrocytic functionality. Nbs1 levels were dramatically reduced in adult astrocyte isolated from Nbs1-CNS-Δ mice, suggesting a major role in cerebellar pathology. In order to investigate the effect of Nbs1 deletion on astrocyte activity, we investigated glutamine synthetase levels in astrocyte and discovered 40% reduction as compared to WT. Furthermore, we found a significant reduction in the secretion of neurotrophic factors, such as brain-derived neurotrophic factor and neurotrophin 3. Understanding the contribution of malfunctioning astrocytes to the etiology of NBS can elucidate a hitherto unknown aspect of this disorder.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adelman CA, De S, Petrini JH (2009) Rad50 is dispensable for the maintenance and viability of postmitotic tissues. Mol Cell Biol 29:483–492
Assaf Y, Galron R, Shapira I, Nitzan A, Blumenfeld-Katzir T, Solomon AS, Holdengreber V, Wang ZQ, Shiloh Y, Barzilai A (2008) MRI evidence of white matter damage in a mouse model of Nijmegen breakage syndrome. Exp Neurol 209:181–191
Baranes K, Raz-Prag D, Nitzan A, Galron R, Ashery-Padan R, Rotenstreich Y, Assaf Y, Shiloh Y, Wang ZQ, Barzilai A, Solomon AS (2009) Conditional inactivation of the NBS1 gene in the mouse central nervous system leads to neurodegeneration and disorganization of the visual system. Exp Neurol 218:24–32
Barzilai A, Biton S, Shiloh Y (2008) The role of the DNA damage response in neuronal development, organization and maintenance. DNA Repair (Amst) 7:1010–1027
Bellamy TC (2006) Interactions between Purkinje neurones and Bergmann glia. Cerebellum 5:116–126
Biton S, Barzilai A, Shiloh Y (2008) The neurological phenotype of ataxia-telangiectasia: solving a persistent puzzle. DNA Repair (Amst) 7:1028–1038
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Cardona AE, Huang D, Sasse ME, Ransohoff RM (2006) Isolation of murine microglial cells for RNA analysis or flow cytometry. Nat Protoc 1:1947–1951
Clement AM, Nguyen MD, Roberts EA, Garcia ML, Boillee S, Rule M, McMahon AP, Doucette W, Siwek D, Ferrante RJ, Brown RH Jr, Julien JP, Goldstein LS, Cleveland DW (2003) Wild-type nonneuronal cells extend survival of SOD1 mutant motor neurons in ALS mice. Science 302:113–117
Colon-Ramos DA, Shen K (2008) Cellular conductors: glial cells as guideposts during neural circuit development. PLoS Biol 6:e112
Digweed M, Sperling K (2004) Nijmegen breakage syndrome: clinical manifestation of defective response to DNA double-strand breaks. DNA Repair (Amst) 3:1207–1217
El Khoury J, Toft M, Hickman SE, Means TK, Terada K, Geula C, Luster AD (2007) Ccr2 deficiency impairs microglial accumulation and accelerates progression of Alzheimer-like disease. Nat Med 13:432–438
Falck J, Coates J, Jackson SP (2005) Conserved modes of recruitment of ATM, ATR and DNA-PKcs to sites of DNA damage. Nature 434:605–611
Farfara D, Lifshitz V, Frenkel D (2008) Neuroprotective and neurotoxic properties of glial cells in the pathogenesis of Alzheimer's disease. J Cell Mol Med 12:762–780
Fernandez AM, Carro EM, Lopez-Lopez C, Torres-Aleman I (2005) Insulin-like growth factor I treatment for cerebellar ataxia: addressing a common pathway in the pathological cascade? Brain Res Brain Res Rev 50:134–141
Frappart PO, Tong WM, Demuth I, Radovanovic I, Herceg Z, Aguzzi A, Digweed M, Wang ZQ (2005) An essential function for NBS1 in the prevention of ataxia and cerebellar defects. Nat Med 11:538–544
Giaume C, Kirchhoff F, Matute C, Reichenbach A, Verkhratsky A (2007) Glia: the fulcrum of brain diseases. Cell Death Differ 14:1324–1335
Gros-Louis F, Gaspar C, Rouleau GA (2006) Genetics of familial and sporadic amyotrophic lateral sclerosis. Biochim Biophys Acta 1762:956–972
Hanisch UK, Kettenmann H (2007) Microglia: active sensor and versatile effector cells in the normal and pathologic brain. Nat Neurosci 10:1387–1394
Hardin-Pouzet H, Krakowski M, Bourbonniere L, Didier-Bazes M, Tran E, Owens T (1997) Glutamate metabolism is down-regulated in astrocytes during experimental allergic encephalomyelitis. Glia 20:79–85
Harlow E, Lane D (1988) Antibodies: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Hickman SE, Allison EK, El Khoury J (2008) Microglial dysfunction and defective beta-amyloid clearance pathways in aging Alzheimer's disease mice. J Neurosci 28:8354–8360
Huang DR, Wang J, Kivisakk P, Rollins BJ, Ransohoff RM (2001) Absence of monocyte chemoattractant protein 1 in mice leads to decreased local macrophage recruitment and antigen-specific T helper cell type 1 immune response in experimental autoimmune encephalomyelitis. J Exp Med 193:713–726
Ilieva H, Polymenidou M, Cleveland DW (2009) Non-cell autonomous toxicity in neurodegenerative disorders: ALS and beyond. J Cell Biol 187:761–772
Julien JP, Kriz J (2006) Transgenic mouse models of amyotrophic lateral sclerosis. Biochim Biophys Acta 1762:1013–1024
Kalehua AN, Nagel JE, Whelchel LM, Gides JJ, Pyle RS, Smith RJ, Kusiak JW, Taub DD (2004) Monocyte chemoattractant protein-1 and macrophage inflammatory protein-2 are involved in both excitotoxin-induced neurodegeneration and regeneration. Exp Cell Res 297:197–211
Kettenmann H, Verkhratsky A (2008) Neuroglia: the 150 years after. Trends Neurosci 31:653–659
Kim J, Wong PK (2009) Oxidative stress is linked to ERK1/2-p16 signaling-mediated growth defect in ATM-deficient astrocytes. J Biol Chem 284(21):14396–14404
Ma DK, Ming GL, Song H (2005) Glial influences on neural stem cell development: cellular niches for adult neurogenesis. Curr Opin Neurobiol 15:514–520
Nimmerjahn A (2009) Astrocytes going live: advances and challenges. J Physiol 587:1639–1647
Streit WJ (2006) Microglial senescence: does the brain's immune system have an expiration date? Trends Neurosci 29:506–510
Sudarov A, Joyner AL (2007) Cerebellum morphogenesis: the foliation pattern is orchestrated by multi-cellular anchoring centers. Neural Dev 2:26
Swanson RA, Ying W, Kauppinen TM (2004) Astrocyte influences on ischemic neuronal death. Curr Mol Med 4:193–205
Taylor AM, Groom A, Byrd PJ (2004) Ataxia-telangiectasia-like disorder (ATLD)—its clinical presentation and molecular basis. DNA Repair (Amst) 3:1219–1225
Uziel T, Lerenthal Y, Moyal L, Andegeko Y, Mittelman L, Shiloh Y (2003) Requirement of the MRN complex for ATM activation by DNA damage. EMBO J 22:5612–5621
Van Den Bosch M, Bree RT, Lowndes NF (2003) The MRN complex: coordinating and mediating the response to broken chromosomes. EMBO Rep 4:844–849
Waltes R, Kalb R, Gatei M, Kijas AW, Stumm M, Sobeck A, Wieland B, Varon R, Lerenthal Y, Lavin MF, Schindler D, Dork T (2009) Human RAD50 deficiency in a Nijmegen breakage syndrome-like disorder. Am J Hum Genet 84:605–616
Zhang C, Shen JK, Lam TT, Zeng HY, Chiang SK, Yang F, Tso MO (2005) Activation of microglia and chemokines in light-induced retinal degeneration. Mol Vis 11:887–895
Acknowledgment
This work is partially supported by grants from the HFSP organization, Dana Foundation and ISF‐Legacy Heritage Biomedical Science Partnership 862/09 (to D.F.) and by the A-T Children Project, the German-Israeli Foundation (to A.B., Z-Q. W, and Y.S.), the USA–Israel Binational Science Foundation, and the Israeli Science Foundation (to A.B.). Z.-Q.W. is supported by the Association for International Cancer Research (AICR), UK and by the Deutschen Forschungsgemeinschaft (DFG), Germany. Work in the laboratory of YS is supported by research grants from the A-T Medical Research Foundation, The Israel Cancer Research Fund, and the A-T Ease Foundation. YS is an Israel Cancer Research Fund Research Professor.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Galron, R., Gruber, R., Lifshitz, V. et al. Astrocyte Dysfunction Associated with Cerebellar Attrition in a Nijmegen Breakage Syndrome Animal Model. J Mol Neurosci 45, 202–211 (2011). https://doi.org/10.1007/s12031-011-9494-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12031-011-9494-6