Skip to main content
SpringerLink
Log in

Upregulation of Vesicular Glutamate Transporter 2 and STAT3 Activation in the Spinal Cord of Mice Receiving 3,3′-Iminodipropionitrile

  • ORIGINAL ARTICLE
  • Published:
Neurotoxicity Research Aims and scope Submit manuscript

Abstract

Chronic administration of 3,3′-iminodipropionitrile (IDPN) causes axonal impairment. Although controversy still remains, it has been suggested that IDPN intoxication mimics the axonopathy of amyotrophic lateral sclerosis (ALS). Interestingly, recent studies including our own showed that signal transducer and activator of transcription 3 (STAT3) in spinal α-motoneurons was activated in both IDPN-treated mice and SOD1 G93A mice, a genetic model of familial ALS. Because activation of STAT3 occurs in response to various stimuli, such as axonal injury, ischemia, and excessive glutamate, here we focused on a potential link between phosphorylated STAT3 (pSTAT3, an active form) and vesicular glutamate transporter 2 (VGluT2, a regulator of glutamate storage and release) in IDPN-treated mice and SOD1 G93A mice. Impairment of axonal transport was confirmed by western blot analysis: the expression levels of phosphorylated neurofilament H were elevated in both models. As shown in SOD1 G93A mice, the expression frequencies of VGluT2 in synaptophysin-positive (SYP)+ presynaptic terminals around spinal α-motoneurons were significantly higher in IDPN-treated mice than in vehicle controls. The coverages of spinal α-motoneurons by VGluT2+ presynaptic terminals were more elevated around pSTAT3+ cells than around pSTAT3− cells in IDPN-treated mice and SOD1 G93A mice. Considering that excessive glutamate is shown to be involved in axonal impairment and STAT3 activation, the present results suggest that IDPN-induced upregulation of VGluT2 may result in an increase in glutamate, which might cause axonopathy and induction of pSTAT3. The link between upregulation of VGluT2 and activation of STAT3 via glutamate may represent a common pathological feature of IDPN-treated mice and SOD1 G93A mice.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Ackerley S, Grierson AJ, Brownlees J, Thornhill P, Anderton BH, Leigh PN, Shaw CE, Miller CC (2000) Glutamate slows axonal transport of neurofilaments in transfected neurons. J Cell Biol 150(1):165–176

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Aguilera P, Ortega A (1999) Stat3 participates in the metabotropic glutamate signaling pathway in Bergmann glial cells. Neurochem Res 24(8):981–986

    Article  CAS  PubMed  Google Scholar 

  • Beers DR, Henkel JS, Zhao W, Wang J, Appel SH (2008) CD4+ T cells support glial neuroprotection, slow disease progression, and modify glial morphology in an animal model of inherited ALS. Proc Natl Acad Sci U S A 105(40):15558–15563

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bilsland LG, Sahai E, Kelly G, Golding M, Greensmith L, Schiavo G (2010) Deficits in axonal transport precede ALS symptoms in vivo. Proc Natl Acad Sci U S A 107(47):20523–20528

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blakely RD, Edwards RH (2012) Vesicular and plasma membrane transporters for neurotransmitters. Cold Spring Harb Perspect Biol 4(2)

  • Chernigovskaya EV, Korotkov AA, Nikitina LS, Dorofeeva NA, Glazova MV (2015) The expression and distribution of seizure-related and synaptic proteins in the insular cortex of rats genetically prone to audiogenic seizures. Neurol Res 37(12):1108–1117

    Article  CAS  PubMed  Google Scholar 

  • Chiba T, Yamada M, Aiso S (2009) Targeting the JAK2/STAT3 axis in Alzheimer’s disease. Expert Opin Ther Targets 13(10):1155–1167

    Article  CAS  PubMed  Google Scholar 

  • Chou SM, Wang HS, Taniguchi A, Bucala R (1998) Advanced glycation endproducts in neurofilament conglomeration of motoneurons in familial and sporadic amyotrophic lateral sclerosis. Mol Med 4(5):324–332

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cudkowicz ME, Titus S, Kearney M, Yu H, Sherman A, Schoenfeld D, Hayden D, Shui A, Brooks B, Conwit R, Felsenstein D, Greenblatt DJ, Keroack M, Kissel JT, Miller R, Rosenfeld J, Rothstein JD, Simpson E, Tolkoff-Rubin N, Zinman L, Shefner JM, Investigators CS (2014) Safety and efficacy of ceftriaxone for amyotrophic lateral sclerosis: a multi-stage, randomised, double-blind, placebo-controlled trial. Lancet Neurol 13(11):1083–1091

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Daniels RW, Miller BR, DiAntonio A (2011) Increased vesicular glutamate transporter expression causes excitotoxic neurodegeneration. Neurobiol Dis 41(2):415–420

    Article  CAS  PubMed  Google Scholar 

  • Delay J, Pichot P, Thuillier J, Marquiset JP (1952) Effect of aminodipropionitrile on motor behavior of the white mouse. C R Seances Soc Biol Fil 146(7–8):533–534

    CAS  PubMed  Google Scholar 

  • Dolcet X, Soler RM, Gould TW, Egea J, Oppenheim RW, Comella JX (2001) Cytokines promote motoneuron survival through the Janus kinase-dependent activation of the phosphatidylinositol 3-kinase pathway. Mol Cell Neurosci 18(6):619–631

    Article  CAS  PubMed  Google Scholar 

  • Dziennis S, Alkayed NJ (2008) Role of signal transducer and activator of transcription 3 in neuronal survival and regeneration. Rev Neurosci 19(4–5):341–361

    CAS  PubMed  PubMed Central  Google Scholar 

  • Friese A, Kaltschmidt JA, Ladle DR, Sigrist M, Jessell TM, Arber S (2009) Gamma and alpha motor neurons distinguished by expression of transcription factor Err3. Proc Natl Acad Sci U S A 106(32):13588–13593

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fujimoto H, Konno K, Watanabe M, Jinno S (2017) Late postnatal shifts of parvalbumin and nitric oxide synthase expression within the GABAergic and glutamatergic phenotypes of inferior colliculus neurons. J Comp Neurol 525(4):868–884

    Article  CAS  PubMed  Google Scholar 

  • Gold BG (1987) The pathophysiology of proximal neurofilamentous giant axonal swellings: implications for the pathogenesis of amyotrophic lateral sclerosis. Toxicology 46(2):125–139

    Article  CAS  PubMed  Google Scholar 

  • Griffin JW, Hoffman PN, Clark AW, Carroll PT, Price DL (1978) Slow axonal transport of neurofilament proteins: impairment of beta,beta'-iminodipropionitrile administration. Science 202(4368):633–635

    Article  CAS  PubMed  Google Scholar 

  • Gundersen HJ (1986) Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson J Microsc 143(Pt 1):3–45

    Article  CAS  PubMed  Google Scholar 

  • Gundersen HJ, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147(Pt 3):229–263

    Article  CAS  PubMed  Google Scholar 

  • Guo H, Lai L, Butchbach ME, Stockinger MP, Shan X, Bishop GA, Lin CL (2003) Increased expression of the glial glutamate transporter EAAT2 modulates excitotoxicity and delays the onset but not the outcome of ALS in mice. Hum Mol Genet 12(19):2519–2532

    Article  CAS  PubMed  Google Scholar 

  • Gurney ME, Pu H, Chiu AY, Dal Canto MC, Polchow CY, Alexander DD, Caliendo J, Hentati A, Kwon YW, Deng HX et al (1994) Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation. Science 264(5166):1772–1775

    Article  CAS  PubMed  Google Scholar 

  • Haas CA, Hofmann HD, Kirsch M (1999) Expression of CNTF/LIF-receptor components and activation of STAT3 signaling in axotomized facial motoneurons: evidence for a sequential postlesional function of the cytokines. J Neurobiol 41(4):559–571

    Article  CAS  PubMed  Google Scholar 

  • Hedreen JC (1998) Lost caps in histological counting methods. Anat Rec 250(3):366–372

    Article  CAS  PubMed  Google Scholar 

  • Hiruma H, Katakura T, Takahashi S, Ichikawa T, Kawakami T (2003) Glutamate and amyloid beta-protein rapidly inhibit fast axonal transport in cultured rat hippocampal neurons by different mechanisms. J Neurosci 23(26):8967–8977

    Article  CAS  PubMed  Google Scholar 

  • Jinno S, Aika Y, Fukuda T, Kosaka T (1998) Quantitative analysis of GABAergic neurons in the mouse hippocampus, with optical disector using confocal laser scanning microscope. Brain Res 814(1–2):55–70

    Article  CAS  PubMed  Google Scholar 

  • Langlais PJ, Huang PC, Gabay S (1975) Regional neurochemical studies on the effect of beta, beta'-iminodipropionitrile (IDPN) in the rat. J Neurosci Res 1(5–6):419–435

    Article  CAS  PubMed  Google Scholar 

  • Lee N, Neitzel KL, Devlin BK, MacLennan AJ (2004) STAT3 phosphorylation in injured axons before sensory and motor neuron nuclei: potential role for STAT3 as a retrograde signaling transcription factor. J Comp Neurol 474(4):535–545

    Article  CAS  PubMed  Google Scholar 

  • Liang Z, Wu G, Fan C, Xu J, Jiang S, Yan X, Di S, Ma Z, Hu W, Yang Y (2016) The emerging role of signal transducer and activator of transcription 3 in cerebral ischemic and hemorrhagic stroke. Prog Neurobiol 137:1–16

    Article  CAS  PubMed  Google Scholar 

  • Liu KX, Edwards B, Lee S, Finelli MJ, Davies B, Davies KE, Oliver PL (2015) Neuron-specific antioxidant OXR1 extends survival of a mouse model of amyotrophic lateral sclerosis. Brain 138(Pt 5):1167–1181

    Article  PubMed  PubMed Central  Google Scholar 

  • Lu JQ, Power C, Blevins G, Giuliani F, Yong VW (2013) The regulation of reactive changes around multiple sclerosis lesions by phosphorylated signal transducer and activator of transcription. J Neuropathol Exp Neurol 72(12):1135–1144

    Article  CAS  PubMed  Google Scholar 

  • Mancuso R, Olivan S, Mancera P, Pasten-Zamorano A, Manzano R, Casas C, Osta R, Navarro X (2012) Effect of genetic background on onset and disease progression in the SOD1-G93A model of amyotrophic lateral sclerosis. Amyotroph Lateral Scler 13(3):302–310

    Article  CAS  PubMed  Google Scholar 

  • McGregor G, Irving AJ, Harvey J (2017) Canonical JAK-STAT signaling is pivotal for long-term depression at adult hippocampal temporoammonic-CA1 synapses. Faseb j 31(8):3449–3466

    Article  CAS  PubMed  Google Scholar 

  • Mehta A, Prabhakar M, Kumar P, Deshmukh R, Sharma PL (2013) Excitotoxicity: bridge to various triggers in neurodegenerative disorders. Eur J Pharmacol 698(1–3):6–18

    Article  CAS  PubMed  Google Scholar 

  • Mendonca DM, Chimelli L, Martinez AM (2005) Quantitative evidence for neurofilament heavy subunit aggregation in motor neurons of spinal cords of patients with amyotrophic lateral sclerosis. Braz J Med Biol Res 38(6):925–933

    Article  CAS  PubMed  Google Scholar 

  • Moechars D, Weston MC, Leo S, Callaerts-Vegh Z, Goris I, Daneels G, Buist A, Cik M, van der Spek P, Kass S, Meert T, D'Hooge R, Rosenmund C, Hampson RM (2006) Vesicular glutamate transporter VGLUT2 expression levels control quantal size and neuropathic pain. J Neurosci 26(46):12055–12066

    Article  CAS  PubMed  Google Scholar 

  • Munoz DG, Greene C, Perl DP, Selkoe DJ (1988) Accumulation of phosphorylated neurofilaments in anterior horn motoneurons of amyotrophic lateral sclerosis patients. J Neuropathol Exp Neurol 47(1):9–18

    Article  CAS  PubMed  Google Scholar 

  • Murphy-Royal C, Dupuis J, Groc L, Oliet SH (2017) Astroglial glutamate transporters in the brain: regulating neurotransmitter homeostasis and synaptic transmission. J Neurosci Res 95(11):2140–2151

    Article  CAS  PubMed  Google Scholar 

  • Ni B, Rosteck PR, Nadi NS, Paul SM (1994) Cloning and expression of a cDNA encoding a brain-specific Na(+)-dependent inorganic phosphate cotransporter. Proc Natl Acad Sci U S A 91(12):5607–5611

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nicolas CS, Amici M, Bortolotto ZA, Doherty A, Csaba Z, Fafouri A, Dournaud P, Gressens P, Collingridge GL, Peineau S (2013) The role of JAK-STAT signaling within the CNS. Jakstat 2(1):e22925

    PubMed  PubMed Central  Google Scholar 

  • Ohgomori T, Yamada J, Takeuchi H, Kadomatsu K, Jinno S (2016) Comparative morphometric analysis of microglia in the spinal cord of SOD1 G93A transgenic mouse model of amyotrophic lateral sclerosis. Eur J Neurosci 43(10):1340–1351

    Article  PubMed  Google Scholar 

  • Ohgomori T, Yamasaki R, Takeuchi H, Kadomatsu K, Kira JI, Jinno S (2017a) Differential activation of neuronal and glial STAT3 in the spinal cord of the SOD1G93A mouse model of amyotrophic lateral sclerosis. Eur J Neurosci 46(4):2001–2014

    Article  PubMed  Google Scholar 

  • Ohgomori T, Yamasaki R, Takeuchi H, Kadomatsu K, Kira JI, Jinno S (2017b) Differential involvement of vesicular and glial glutamate transporters around spinal alpha-motoneurons in the pathogenesis of SOD1G93A mouse model of amyotrophic lateral sclerosis. Neuroscience 356:114–124

    Article  CAS  PubMed  Google Scholar 

  • Park KW, Nozell SE, Benveniste EN (2012) Protective role of STAT3 in NMDA and glutamate-induced neuronal death: negative regulatory effect of SOCS3. PLoS One 7(11):e50874

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rothstein JD, Van Kammen M, Levey AI, Martin LJ, Kuncl RW (1995) Selective loss of glial glutamate transporter GLT-1 in amyotrophic lateral sclerosis. Ann Neurol 38(1):73–84

    Article  CAS  PubMed  Google Scholar 

  • Saba L, Viscomi MT, Caioli S, Pignataro A, Bisicchia E, Pieri M, Molinari M, Ammassari-Teule M, Zona C (2016) Altered functionality, morphology, and vesicular glutamate transporter expression of cortical motor neurons from a presymptomatic mouse model of amyotrophic lateral sclerosis. Cereb Cortex 26(4):1512–1528

    Article  CAS  PubMed  Google Scholar 

  • Schweizer U, Gunnersen J, Karch C, Wiese S, Holtmann B, Takeda K, Akira S, Sendtner M (2002) Conditional gene ablation of Stat3 reveals differential signaling requirements for survival of motoneurons during development and after nerve injury in the adult. J Cell Biol 156(2):287–297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shibata N, Kakita A, Takahashi H, Ihara Y, Nobukuni K, Fujimura H, Sakoda S, Sasaki S, Iwata M, Morikawa S, Hirano A, Kobayashi M (2009) Activation of signal transducer and activator of transcription-3 in the spinal cord of sporadic amyotrophic lateral sclerosis patients. Neurodegener Dis 6(3):118–126

    Article  CAS  PubMed  Google Scholar 

  • Shibata N, Yamamoto T, Hiroi A, Omi Y, Kato Y, Kobayashi M (2010) Activation of STAT3 and inhibitory effects of pioglitazone on STAT3 activity in a mouse model of SOD1-mutated amyotrophic lateral sclerosis. Neuropathology 30(4):353–360

    Article  PubMed  Google Scholar 

  • Soler-Martin C, Boadas-Vaello P, Verdu E, Garcia N, Llorens J (2014) Chronic proximal axonopathy in rats is associated with long-standing neurofilament depletion in neuromuscular junctions and behavioral deficits. J Neuropathol Exp Neurol 73(6):568–579

    Article  PubMed  Google Scholar 

  • Takamori S, Rhee JS, Rosenmund C, Jahn R (2000) Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons. Nature 407(6801):189–194

    Article  CAS  PubMed  Google Scholar 

  • Takamori S, Rhee JS, Rosenmund C, Jahn R (2001) Identification of differentiation-associated brain-specific phosphate transporter as a second vesicular glutamate transporter (VGLUT2). J Neurosci 21(22):RC182

    Article  CAS  PubMed  Google Scholar 

  • Takanashi T (1993) Free amino acids and activities of folate-derivative converting enzymes in nervous system of rats administered with beta, beta'-iminodipropionitrile. Rinsho Shinkeigaku 33(8):862–869

    CAS  PubMed  Google Scholar 

  • Tariq M, Khan HA, Siddiquei MM, Al Moutaery K, Al DS (2007) Protective effect of hydrocortisone on iminodipropionitrile-induced neurotoxicity in rats. Basic Clin Pharmacol Toxicol 100(3):176–181

    Article  CAS  PubMed  Google Scholar 

  • Tashiro T, Imai R, Komiya Y (1994) Early effects of beta,beta'-iminodipropionitrile on tubulin solubility and neurofilament phosphorylation in the axon. J Neurochem 63(1):291–300

    Article  CAS  PubMed  Google Scholar 

  • Toth E, Lajtha A, Vizi ES (1997) Effects of iminodipropionitrile on cerebral amino acid levels. Brain Res Bull 44(6):715–718

    Article  CAS  PubMed  Google Scholar 

  • Vivanco F, Ramos F, Jimenez-Diaz C (1966) Determination of gamma-aminobutyric acid and other free amino acids in whole brains of rats poisoned with beta, beta'-iminodipropionitrile and alpha, gamma-diaminobutyric acid with, or without, administration of thyroxine. J Neurochem 13(12):1461–1467

    Article  CAS  PubMed  Google Scholar 

  • Wang DV, Viereckel T, Zell V, Konradsson-Geuken A, Broker CJ, Talishinsky A, Yoo JH, Galinato MH, Arvidsson E, Kesner AJ, Hnasko TS, Wallen-Mackenzie A, Ikemoto S (2017) Disrupting glutamate co-transmission does not affect acquisition of conditioned behavior reinforced by dopamine neuron activation. Cell Rep 18(11):2584–2591

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang ZT, Yu G, Wang HS, Yi SP, Su RB, Gong ZH (2015) Changes in VGLUT2 expression and function in pain-related supraspinal regions correlate with the pathogenesis of neuropathic pain in a mouse spared nerve injury model. Brain Res 1624:515–524

    Article  CAS  PubMed  Google Scholar 

  • Williamson TL, Cleveland DW (1999) Slowing of axonal transport is a very early event in the toxicity of ALS-linked SOD1 mutants to motor neurons. Nat Neurosci 2(1):50–56

    Article  CAS  PubMed  Google Scholar 

  • Wilson NR, Kang J, Hueske EV, Leung T, Varoqui H, Murnick JG, Erickson JD, Liu G (2005) Presynaptic regulation of quantal size by the vesicular glutamate transporter VGLUT1. J Neurosci 25(26):6221–6234

    Article  CAS  PubMed  Google Scholar 

  • Wootz H, Enjin A, Wallen-Mackenzie A, Lindholm D, Kullander K (2010) Reduced VGLUT2 expression increases motor neuron viability in Sod1(G93A) mice. Neurobiol Dis 37(1):58–66

    Article  CAS  PubMed  Google Scholar 

  • Yamada J, Nakanishi H, Jinno S (2011) Differential involvement of perineuronal astrocytes and microglia in synaptic stripping after hypoglossal axotomy. Neuroscience 182:1–10

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, Fauser U, Schluesener HJ (2008) Dexamethasone suppresses infiltration of RhoA+ cells into early lesions of rat traumatic brain injury. Acta Neuropathol 115(3):335–343

    Article  CAS  PubMed  Google Scholar 

  • Zhu Q, Lindenbaum M, Levavasseur F, Jacomy H, Julien JP. 1998. Disruption of the NF-H gene increases axonal microtubule content and velocity of neurofilament transport: relief of axonopathy resulting from the toxin beta,beta'-iminodipropionitrile. J Cell Biol 143(1):183–193

Download references

Acknowledgements

The authors thank Ms. Machiko Endoh for her technical and secretarial assistance. We also thank Editage (www.editage.jp) for English language editing. We also appreciate the technical assistance from The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences.

Author information

Authors and Affiliations

  1. Department of Anatomy and Neuroscience, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Tomohiro Ohgomori & Shozo Jinno

  2. Department of Neurology, Neurological Institute, Graduate School of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Ryo Yamasaki & Jun-ichi Kira

Authors
  1. Tomohiro Ohgomori
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ryo Yamasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun-ichi Kira
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shozo Jinno
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Study concept and design: S.J. Acquisition of data and statistical analysis: T.O. Drafting of the manuscript and obtained funding: T.O. and S.J. Administrative, technical, and material support: R.Y. and J.K.

Corresponding author

Correspondence to Shozo Jinno.

Ethics declarations

The Committee of Ethics on Animal Experiment in Kyushu University (No. A28-218-0) approved the procedure.

Conflict of Interest

The authors declare that they have no conflicts of interest.

Grant Information

This work was supported in part by JSPS KAKENHI Grant Numbers 25860234 (to T.O.), 17K00858 (to T.O.), 15H04267 (to S.J.), and Grants-in-Aid for Scientific Research on the Innovative Areas of “Deciphering sugar chain-based signals regulating integrative neuronal functions” (26110714 to S.J.).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ohgomori, T., Yamasaki, R., Kira, Ji. et al. Upregulation of Vesicular Glutamate Transporter 2 and STAT3 Activation in the Spinal Cord of Mice Receiving 3,3′-Iminodipropionitrile. Neurotox Res 33, 768–780 (2018). https://doi.org/10.1007/s12640-017-9822-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12640-017-9822-x

Keywords

Search

Navigation