Supplementary Table 1. Evidence for glutamatergic system dysfunction in MS
Sample
Method
Finding
Reference
CSF (RRMS)
HPLC
↑
Stover et al.1
CSF (RRMS)
HPLC
↑
Sarchielli et al.2
CSF
Proteomics
↑
Pieragostino et al.3
CP, AP, WM (RRMS,
SPMS, PPMS)
Normal WM (RRMS,
SPMS, PPMS)
Normal WM and GM (all
types of MS)
WM (SP-MS)
MRS 3T
No change in CP
↑normal-appearing WM, AP
↑
Srinivasan et al.4
↑
Baranzini et al. 6
MRS
↓
Mac Millan et al. 7
GM (RRMS)
MRI
↓Small
Muhlert et al.8
CSF (unspecified MS)
HPLC
↓
Qureshi & Baig9
IHC
↑Glutaminase
Werner et al.10
IHC
↓GS and GDH
Werner et al.10
IHC
↑ GS and GDH
Newcombe et al.11
Proteomics
↑GDH
Han et al.12
OLG of WM AP and CP
(PPMS)
OLG of WM AP
IHC
↓EAAT1 and GLT1
Werner et al.10
IHC
↓EAAT1 and GLT1
Pitt et al.13
Optic nerve (RRMS,
SPMS, PPMS)
Cortical lesions (RRMS,
SPMS)
Spinal cord (SPMS,
PPMS)
IHC, WB,
qPCR
IHC
↑EAAT1 and GLT1
↓EAAT1 and GLT1
Vallejo-Illarramendi
et al.14
Vercellino et al.15
IHC
microarray
WB
IHC
↑ EAAT1
Lieury et al.16
↑EAAC1
Newcombe et al.11
WB, IHC,
qPCR
↑cystine/Glu antiporter
Pampliega et al.17
Uptake
↑ in glial vesicles
Vallejo-Illarramendi
et al.14
Non active WM, CP and
AP (SPMS, PPMS)
OLG AP
IHC
Geurts et al.18
IHC
↑mGluR1 (dystrophic axon); mGluR5 and
mGluR2/3 (astrog)
↑AMPAR GluR1
OLG CP
IHC
↓ AMPAR GluR1
Microglia AP
Astroglia and dystrophic
axon
Endothelial cells and
dystrophic axon
Cerebellum (SPMS,
PPMS, RRMS)
CAP and AP (SPMS,
PPMS, RRMS)
CP (RRMS)
IHC
IHC
↑ AMPAR GluR2
↑mGluR1,mGluR2/3, mGluR5
IHC
↑ mGluR5–7
IHC
↓ mGluR1 and↑ mGluR5
Fazio et al.19
Proteomics
↑ mGluR2 in CAP ↑ mGluR3 in AP
Han et al.12
WB
↑ AMPAR GluR2
Zhai et al.20
Glutamate level
MRI and
MRS
MRI
Vrenken et al.5
Glutamate-metabolizing enzymes
Macrophages/microglia
WM (PPMS)
OLG of WM AP and CP
Glutamate
(PPMS)
degradation
Microglia and astroglia
AP
CP and AP (SPMS,
PPMS)
Glutamate transporters/uptake
Glutamate synthesis
Glutamate
transporters
Microglia and astroglia
AP
Spinal cord leukocytes
and Optic nerve (RRMS,
SPMS)
Optic nerve (PPMS,
Glutamate uptake
SPMS RRMS)
Glutamate receptor (GluR) expression
Newcombe et al.11
Abbreviations: AMPAR, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor; AP, active plaque; CAP, chronic active
plaque; CP, chronic plaque; CSF, cerebrospinal fluid; EAAC1, excitatory amino-acid carrier 1; GDH, glu dehydrogenase; EAAT1,
glutamate aspartate transporter; GM, grey matter; GS, glutamine synthetase; HPLC, high-performance liquid chromatography;
IHC, immunohistochemistry; mGluR, metabotropic glutamate receptor; MS, multiple sclerosis; MRS, magnetic resonance
spectroscopy; NMDAR, N-methyl-D-aspartate receptor; OLG, oligodendrocyte; PP, primary progressive; qPCR, quantitative
PCR; RR, relapsing–remitting; SP-secondary progressive; WB, western blot; WM, white matter.
1. Stover, J. F. et al. Neurotransmitters in cerebrospinal fluid reflect pathological activity. Eur. J. Clin. Invest. 27, 1038–1043
(1997).
2. Sarchielli, P., Greco, L., Floridi, A., Floridi, A. & Gallai, V. Excitatory amino acids and multiple sclerosis: evidence from
cerebrospinal fluid. Arch. Neurol. 60, 1082–1088 (2003).
3. Pieragostino, D. et al. An integrated metabolomics approach for the research of new cerebrospinal fluid biomarkers of
multiple sclerosis. Mol. Biosyst. 11, 1563–1572 (2015).
4. Srinivasan, R., Sailasuta, N., Hurd, R., Nelson, S. & Pelletier, D. Evidence of elevated glutamate in multiple sclerosis
using magnetic resonance spectroscopy at 3 T. Brain 128, 1016–1025 (2005).
5. Vrenken, H. et al. MR spectroscopic evidence for glial increase but not for neuro-axonal damage in MS normal-appearing
white matter. Magn. Reson. Med. 53, 256–266 (2005).
6. Baranzini, S. E. et al. Genetic variation influences glutamate concentrations in brains of patients with multiple sclerosis.
Brain 133, 2603-2611 (2010).
7. MacMillan, E. L. et al. Progressive multiple sclerosis exhibits decreasing glutamate and glutamine over two years. Mult.
Scler. (2015). doi:10.1177/1352458515586086
8. Muhlert, N. et al. Memory in multiple sclerosis is linked to glutamate concentration in grey matter regions. J. Neurol.
Neurosurg. Psychiatry 85, 834–840 (2014).
9. Qureshi, G. A. & Baig, M. S. Quantitation of free amino acids in biological samples by high-performance liquid
chromatography. Application of the method in evaluating amino acid levels in cerebrospinal fluid and plasma of patients
with multiple sclerosis. J. Chromatogr. 459, 237–244 (1988).
10. Werner, P., Pitt, D. & Raine, C. S. Multiple sclerosis: Altered glutamate homeostasis in lesions correlates with
oligodendrocyre and axonal damage. Ann. Neurol. 50, 169–180 (2001).
11. Newcombe, J. et al. Glutamate receptor expression in multiple sclerosis lesions. Brain Pathol. 18, 52–61 (2008).
12. Han, M. H. et al. Proteomic analysis of active multiple sclerosis lesions reveals therapeutic targets. Nature 451, 10761081 (2008).
13. Pitt, D., Nagelmeier, I. E., Wilson, H. C. & Raine, C. S. Glutamate uptake by oligodendrocytes: Implications for
excitotoxicity in multiple sclerosis. Neurology 61, 1113–1120 (2003).
14. Vallejo-Illarramendi, A., Domercq, M., Pérez-Cerdá, F., Ravid, R. & Matute, C. Increased expression and function of
glutamate transporters in multiple sclerosis. Neurobiol. Dis. 21, 154–164 (2006).
15. Vercellino, M. et al. Altered glutamate reuptake in relapsing-remitting and secondary progressive multiple sclerosis
cortex: correlation with microglia infiltration, demyelination, and neuronal and synaptic damage. J. Neuropathol. Exp.
Neurol. 66, 723-729 (2007).
16. Lieury, A. et al. Tissue Remodeling in Periplaque Regions of Multiple Sclerosis Spinal Cord Lesions. Glia 62, 1645–
1658 (2014).
17. Pampliega, O. et al. Increased expression of cystine/glutamate antiporter in multiple sclerosis. J. Neuroinflammation 8,
63 (2011).
18. Geurts, J. J. G. et al. Altered expression patterns of group I and II metabotropic glutamate receptors in multiple sclerosis.
Brain 126, 1755–1766 (2003).
19. Fazio, F. et al. Switch in the expression of mGlu1 and mGlu5 metabotropic glutamate receptors in the cerebellum of
mice developing experimental autoimmune encephalomyelitis and in autoptic cerebellar samples from patients with multiple
sclerosis. Neuropharmacology 55, 491–499 (2008).
20. Zhai, D. et al. Blocking GluR2-GAPDH ameliorates experimental autoimmune encephalomyelitis. Ann. Clin. Transl.
Neurol. 2, 388–400 (2015).