Synaptic currents and transmitter responses in human NT2 neurons differentiated in aggregate culture

Podrygajlo, Grzegorz; Song, Yunping; Schlesinger, Friedrich; Kenn, Klaus; Bicker, Gerd

doi:10.1016/j.neulet.2009.10.092

Cited by 22 publications

(17 citation statements)

References 26 publications

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“…2 L the application of ACM (measured GABA concentration: 8–14 µmol) induced inward current i.e. response consistent with GABA A receptor-mediated depolarization [22], [23]. This response was similar to that measured after application of 50 µM GABA ( Fig.…”

Section: Resultssupporting

confidence: 82%

Combinational Spinal GAD65 Gene Delivery and Systemic GABA-Mimetic Treatment for Modulation of Spasticity

et al. 2012

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BackgroundLoss of GABA-mediated pre-synaptic inhibition after spinal injury plays a key role in the progressive increase in spinal reflexes and the appearance of spasticity. Clinical studies show that the use of baclofen (GABAB receptor agonist), while effective in modulating spasticity is associated with major side effects such as general sedation and progressive tolerance development. The goal of the present study was to assess if a combined therapy composed of spinal segment-specific upregulation of GAD65 (glutamate decarboxylase) gene once combined with systemic treatment with tiagabine (GABA uptake inhibitor) will lead to an antispasticity effect and whether such an effect will only be present in GAD65 gene over-expressing spinal segments.Methods/Principal FindingsAdult Sprague-Dawley (SD) rats were exposed to transient spinal ischemia (10 min) to induce muscle spasticity. Animals then received lumbar injection of HIV1-CMV-GAD65 lentivirus (LVs) targeting ventral α-motoneuronal pools. At 2–3 weeks after lentivirus delivery animals were treated systemically with tiagabine (4, 10, 20 or 40 mg/kg or vehicle) and the degree of spasticity response measured. In a separate experiment the expression of GAD65 gene after spinal parenchymal delivery of GAD65-lentivirus in naive minipigs was studied. Spastic SD rats receiving spinal injections of the GAD65 gene and treated with systemic tiagabine showed potent and tiagabine-dose-dependent alleviation of spasticity. Neither treatment alone (i.e., GAD65-LVs injection only or tiagabine treatment only) had any significant antispasticity effect nor had any detectable side effect. Measured antispasticity effect correlated with increase in spinal parenchymal GABA synthesis and was restricted to spinal segments overexpressing GAD65 gene.Conclusions/SignificanceThese data show that treatment with orally bioavailable GABA-mimetic drugs if combined with spinal-segment-specific GAD65 gene overexpression can represent a novel and highly effective anti-spasticity treatment which is associated with minimal side effects and is restricted to GAD65-gene over-expressing spinal segments.

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Section: Resultssupporting

confidence: 82%

Combinational Spinal GAD65 Gene Delivery and Systemic GABA-Mimetic Treatment for Modulation of Spasticity

et al. 2012

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“…NT2 cells shares many similarity with human embryonic stem cells (Schwartz et al 2005) and differentiation of NT2 cells into neurons has been suggested to resemble vertebrate neurogenesis (Przyborski et al 2000(Przyborski et al , 2003Houldsworth et al 2002;Smith et al 2010). Moreover, fully differentiated NT2 neurons have been shown to express a variety of neurotransmitters in vitro (Guillemain et al 2000;Podrygajlo et al 2009) and form functional synapses (Hartley et al 1999;Podrygajlo et al 2010). Thus, NT2 lines provide a robust and amenable culture system to study certain aspects of neurodevelopmental process that mimic vertebrate neurogenesis.…”

Section: Introductionmentioning

confidence: 99%

Rapid Differentiation of Human Embryonal Carcinoma Stem Cells (NT2) into Neurons for Neurite Outgrowth Analysis

Tegenge

Roloff²,

Bicker³

2011

Cell Mol Neurobiol

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Human neurons derived from stem cells can be employed as in vitro models to predict the potential of neurochemicals affecting neurodevelopmental cellular processes including proliferation, migration, and differentiation. Here, we developed a model of differentiating human neurons from well characterized human embryonal carcinoma stem cells (NT2). NT2 cells were induced to differentiate into neuronal phenotypes after 2 weeks of treatment with retinoic acid in aggregate culture. Nestin positive progenitor cells migrate out of NT2 aggregates and differentiate into βIII-tubulin expressing neuronal cells. Culturing the NT2 cells for an additional 7-14 days resulted in increased percentage of βIII-tubulin expressing cells, elaborating a long neurite that positively stained for axonal marker (Tau) and presynaptic protein (synapsin). We then asked whether neurite outgrowth from NT2 cells is modulated by bioactive chemicals. Since the cAMP/PKA pathway has been widely investigated as a regulator of neurite outgrowth/regeneration in several experimental systems, we used chemical activators and inhibitors of cAMP/PKA pathway in the culture. The adenylyl cyclase activator, forskolin, and cell-permeable analog of cAMP, 8-Br-cAMP increased the percentage of neurite bearing cells and neurite extension. Application of the protein kinase A inhibitors, H-89 and Rp-cAMP, blocked neurite formation. Taken together, NT2 aggregates undergo migration, differentiation, and neurite elaboration and can be used as a model of differentiating human neurons to screen neurochemicals and to understand cellular mechanisms of human nerve cell development.

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“…20 Several other human cell lines mainly derived from embryonic teratocarcinomas (NT2, hNT) are currently used, and they can be directed towards a post-mitotic neuronal phenotype. 21,22 The human H4 neuroglioma cell line has been used to study the oligomerization of intracellular aSyn by fluorescence lifetime imaging (FLIM) for the first time. 23 Moreover, the role of aSyn in the autophagy pathway has been addressed in this cell line.…”

Section: Parkinson's Diseasementioning

confidence: 99%

Studying neurodegenerative diseases in culture models

Schlachetzki

Saliba

Oliveira

2013

Rev. Bras. Psiquiatr.

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Neurodegenerative diseases are pathological conditions that have an insidious onset and chronic progression. Different models have been established to study these diseases in order to understand their underlying mechanisms and to investigate new therapeutic strategies. Although various in vivo models are currently in use, in vitro models might provide important insights about the pathogenesis of these disorders and represent an interesting approach for the screening of potential pharmacological agents. In the present review, we discuss various in vitro and ex vivo models of neurodegenerative disorders in mammalian cells and tissues.

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Synaptic currents and transmitter responses in human NT2 neurons differentiated in aggregate culture

Cited by 22 publications

References 26 publications

Combinational Spinal GAD65 Gene Delivery and Systemic GABA-Mimetic Treatment for Modulation of Spasticity

Combinational Spinal GAD65 Gene Delivery and Systemic GABA-Mimetic Treatment for Modulation of Spasticity

Rapid Differentiation of Human Embryonal Carcinoma Stem Cells (NT2) into Neurons for Neurite Outgrowth Analysis

Studying neurodegenerative diseases in culture models

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