Electrophysiological characterization of Nsc-34 cell line using Microelectrode Array

  • K.R. Sabitha
    Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bengaluru 560 029, India
    Search for articles by this author
  • D. Sanjay
    Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bengaluru 560 029, India
    Search for articles by this author
  • B. Savita
    Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bengaluru 560 029, India
    Search for articles by this author
  • T.R. Raju
    Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bengaluru 560 029, India
    Search for articles by this author
  • T.R. Laxmi
    Corresponding author at: Department of Neurophysiology, Hosur Road, P.B. No. 2900, NIMHANS, Bengaluru 560 029, India.
    Department of Neurophysiology, National Institute of Mental Health and Neurosciences (NIMHANS), Hosur Road, Bengaluru 560 029, India
    Search for articles by this author
Published:September 21, 2016DOI:


      • Electrophysiological properties of NSC-34 cell lines were studied in the present study.
      • NSC 34 cell line has the ability to discharge spontaneously and rhythmically.
      • Mean firing frequency revealed that NSC 34 cell lines have >2 distinct types of neuronal populations.
      • NSC-34 cell line has both principal and interneurons.


      Neurons communicate with each other through intricate network to evolve higher brain functions. The electrical activity of the neurons plays a crucial role in shaping the connectivity. With motor neurons being vulnerable to neurodegenerative diseases, understanding the electrophysiological properties of motor neurons is the need of the hour, in order to comprehend the impairment of connectivity in these diseases. NSC-34 cell line serves as an excellent model to study the properties of motor neurons as they express Choline acetyltransferase (ChAT). Although NSC-34 cell lines have been used to study the effect of various toxicological, neurotrophic and neuroprotective agents, the electrical activity of these cells has not been elucidated. In the current study, we have characterized the electrophysiological properties of NSC-34 cell lines using Micro-Electrode Array (MEA) as a tool. Based on the spike waveform, firing frequency, auto- and cross-correlogram analysis, we demonstrate that NSC-34 cell culture has >2 distinct types of neuronal population: principal excitatory neurons, putative interneurons and unclassified neurons. The presence of interneurons in the NSC-34 culture was characterized by increased expression of GAD-67 markers. Thus, finding an understanding of the electrophysiological properties of different population of neurons in NSC-34 cell line, will have multiple applications in the treatment of neurological disorders.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of the Neurological Sciences
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Cashman N.R.
        • Durham H.D.
        • Blusztajn J.K.
        • Oda K.
        • Tabira T.
        • Shaw I.T.
        • et al.
        Neuroblastoma x spinal cord (NSC) hybrid cell lines resemble developing motor neurons.
        Dev. Dyn. 1992; 194: 209-221
        • Durham H.D.
        • Dahrouge S.
        • Cashman N.R.
        Evaluation of the spinal cord neuron X neuroblastoma hybrid cell line NSC-34 as a model for neurotoxicity testing. [Internet].
        Neurotoxicology. 1993; : 387-395
        • Amano T.
        • Richelson E.
        • Nirenberg M.
        Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites).
        Proc. Natl. Acad. Sci. U. S. A. 1972; 69: 258-263
        • Maier O.
        • Böhm J.
        • Dahm M.
        • Brück S.
        • Beyer C.
        • Johann S.
        Differentiated NSC-34 motoneuron-like cells as experimental model for cholinergic neurodegeneration.
        Neurochem. Int. 2013; 62: 1029-1038
        • Rizzardini M.
        • Mangolini A.
        • Lupi M.
        • Ubezio P.
        • Bendotti C.
        • Cantoni L.
        Low levels of ALS-linked Cu/Zn superoxide dismutase increase the production of reactive oxygen species and cause mitochondrial damage and death in motor neuron-like cells.
        J. Neurol. Sci. 2005; 232: 95-103
        • Vijayalakshmi K.
        • Alladi P.A.
        • Ghosh S.
        • Prasanna V.K.
        • Sagar B.C.
        • Nalini A.
        • et al.
        Evidence of endoplasmic reticular stress in the spinal motor neurons exposed to CSF from sporadic amyotrophic lateral sclerosis patients.
        Neurobiol. Dis. 2011; 41: 695-705
        • Vijayalakshmi K.
        • Alladi P.A.
        • Sathyaprabha T.N.
        • Subramaniam J.R.
        • Nalini A.
        • Raju T.R.
        Cerebrospinal fluid from sporadic amyotrophic lateral sclerosis patients induces degeneration of a cultured motor neuron cell line.
        Brain Res. 2009; 1263: 122-133
        • Kulshreshtha D.
        • Vijayalakshmi K.
        • Alladi P.A.
        • Sathyaprabha T.N.
        • Nalini A.
        • Raju T.R.
        Vascular endothelial growth factor attenuates neurodegenerative changes in the NSC-34 motor neuron cell line induced by cerebrospinal fluid of sporadic amyotrophic lateral sclerosis patients.
        Neurodegener. Dis. 2011; 8: 322-330
        • Fryer H.J.
        • Knox R.J.
        • Strittmatter S.M.
        • Kalb R.G.
        Excitotoxic death of a subset of embryonic rat motor neurons in vitro.
        J. Neurochem. [Internet]. 1999; 72: 500-513
        • Nagaraja T.N.
        • Raju T.R.
        • Gourie-Devi M.
        • Nalini A.
        Neurofilament phosphorylation is enhanced in cultured chick spinal cord neurons exposed to cerebrospinal fluid from amyotrophic lateral sclerosis patients.
        Acta Neuropathol. 1994; 88: 349-352
        • Shobha K.
        • Vijayalakshmi K.
        • Alladi P.A.
        • Nalini A.
        • Sathyaprabha T.N.
        • Raju T.R.
        Altered in-vitro and in-vivo expression of glial glutamate transporter-1 following exposure to cerebrospinal fluid of amyotrophic lateral sclerosis patients.
        J. Neurol. Sci. 2007; 254: 9-16
        • Gross G.W.
        • Rieske E.
        • Kreutzberg G.W.
        • Meyer A.
        A new fixed-array multi-microelectrode system designed for long-term monitoring of extracellular single unit neuronal activity in vitro.
        Neurosci. Lett. 1977; 6: 101-105
        • Potter S.M.
        • DeMarse T.B.
        A new approach to neural cell culture for long-term studies.
        J. Neurosci. Methods. 2001; 110: 17-24
        • Thomas C.A.
        • Springer P.A.
        • Loeb G.E.
        • Berwald-Netter Y.
        • Okun L.M.
        A miniature microelectrode array to monitor the bioelectric activity of cultured cells.
        Exp. Cell Res. 1972; 74: 61-66
        • Berdondini L.
        • Van Der Wal P.D.
        • Guenat O.
        • De Rooij N.F.
        • Koudelka-Hep M.
        • Seitz P.
        • et al.
        High-density electrode array for imaging in vitro electrophysiological activity.
        Biosens. Bioelectron. 2005; 21: 167-174
        • Gray C.M.
        • Maldonado P.E.
        • Wilson M.
        • McNaughton B.
        Tetrodes markedly improve the reliability and yield of multiple single-unit isolation from multi-unit recordings in cat striate cortex.
        J. Neurosci. Methods. 1995; 63: 43-54
        • Wu S.N.
        • Yeh C.C.
        • Huang H.C.
        • So E.C.
        • Lo Y.C.
        Electrophysiological characterization of sodium-activated potassium channels in NG108-15 and NSC-34 motor neuron-like cells.
        Acta Physiol. 2012; 206: 120-134
        • Hammond J.B.
        • Kruger N.J.
        The bradford method for protein quantitation.
        Methods Mol. Biol. 1988; 3: 25-32
        • Trapp B.D.
        • Peterson J.W.
        • Ransohoff R.M.
        • Rudick R.A.
        • Mork S.
        • Bo L.
        • et al.
        Axonal transection in the lesions of multiple sclerosis.
        N. Engl. J. Med. [Internet]. 1998; 338: 278-285
        • Chang A.
        • Nishiyama A.
        • Peterson J.
        • Prineas J.
        • Trapp B.D.
        NG2-positive oligodendrocyte progenitor cells in adult human brain and multiple sclerosis lesions.
        J. Neurosci. 2000; 20: 6404-6412
        • Vijayalakshmi K.
        • Ostwal P.
        • Sumitha R.
        • Shruthi S.
        • Varghese A.M.
        • Mishra P.
        • et al.
        Role of VEGF and VEGFR2 receptor in reversal of ALS-CSF induced degeneration of NSC-34 motor neuron cell line.
        Mol. Neurobiol. 2014;
        • Zschüntzsch J.
        • Schütze S.
        • Hülsmann S.
        • Dibaj P.
        • Neusch C.
        Heterologous expression of a glial kir channel (KCNJ10) in a neuroblastoma spinal cord (NSC-34) cell line.
        Physiol. Res. 2013; 62: 95-105
        • Somogyi P.
        • Klausberger T.
        Defined types of cortical interneurone structure space and spike timing in the hippocampus.
        J. Physiol. [Internet]. 2005; 562: 9-26
        • Freund T.F.
        • Buzsáki G.
        Interneurons of the hippocampus.
        Hippocampus. 1996; 6: 347-470
        • Markram H.
        • Toledo-Rodriguez M.
        • Wang Y.
        • Gupta A.
        • Silberberg G.
        • Wu C.
        Interneurons of the neocortical inhibitory system.
        Nat. Rev. Neurosci. [Internet]. 2004; 5: 793-807
        • Csicsvari J.
        • Jamieson B.
        • Wise K.D.
        • Buzsaki G.
        Mechanisms of gamma oscillations in the hippocampus of the behaving rat.
        Neuron. 2003; 37: 311-322
        • Nishimaru H.
        • Restrepo C.E.
        • Ryge J.
        • Yanagawa Y.
        • Kiehn O.
        Mammalian motor neurons corelease glutamate and acetylcholine at central synapses.
        Proc. Natl. Acad. Sci. U. S. A. 2005; 102: 5245-5249
        • Csicsvari J.
        • Hirase H.
        • Czurko A.
        • Buzsaki G.
        Reliability and state dependence of pyramidal cell-interneuron synapses in the hippocampus: an ensemble approach in the behaving rat.
        Neuron. 1998; 21: 179-189
        • O'Keefe J.
        • Recce M.L.
        Phase relationship between hippocampal place units and the EEG theta rhythm.
        Hippocampus. 1993; 3: 317-330
        • Eggett C.J.
        • Crosier S.
        • Manning P.
        • Cookson M.R.
        • Menzies F.M.
        • McNeil C.J.
        • et al.
        Development and characterisation of a glutamate-sensitive motor neurone cell line.
        J. Neurochem. 2000; 74: 1895-1902
        • Gavazzo P.
        • Vella S.
        • Marchetti C.
        • Nizzari M.
        • Cancedda R.
        • Pagano A.
        Acquisition of neuron-like electrophysiological properties in neuroblastoma cells by controlled expression of NDM29 ncRNA.
        J. Neurochem. 2011 Dec 1; 119: 989-1001
        • Becchetti A.
        • Gullo F.
        • Bruno G.
        • Dossi E.
        • Lecchi M.
        • Wanke E.
        Exact distinction of excitatory and inhibitory neurons in neural networks: a study with GFP-GAD67 neurons optically and electrophysiologically recognized on multielectrode arrays.
        Front. Neural. Circuits. 2012 Sep 6; 6: 63
        • Huang J.
        • Chen J.
        • Wang W.
        • Wei Y.Y.
        • Cai G.H.
        • Tamamaki N.
        • Li Y.Q.
        • Wu S.X.
        Birthdate study of GABAergic neurons in the lumbar spinal cord of the glutamic acid decarboxylase 67-green fluorescent protein knock-in mouse.
        Front. Neuroanat. 2013; 7