Human NT2 neurons express a large variety of neurotransmission phenotypes in vitro

Guillemain, Isabelle; Alonso, Gérard; Patey, Gilles; Privat, Alain; Chaudieu, Isabelle

doi:10.1002/1096-9861(20000703)422:3<380::aid-cne5>3.0.co;2-c

Cited by 94 publications

(76 citation statements)

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“…After treatment with retinoic acid and mitotic inhibitors, these cells develop into polarized, postmitotic, neuron-like cells that share many characteristics with CNS neurons (10). They express functional non-NMDA as well as NMDA-receptors (11), different types of calcium channels (12), and elaborate classical synaptic contacts (13). In this descriptive study, we determined the modes of cell death in NT2-N neurons and its temporal evolution after exposure to various concentrations of bilirubin.…”

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confidence: 99%

Bilirubin Induces Apoptosis and Necrosis in Human NT2-N Neurons

et al. 2005

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Studies on primary cultures of newborn rodent neurons have suggested that neuronal death induced by unconjugated bilirubin (UCB) is mainly apoptotic in nature. We exposed a human teratocarcinoma-derived cell line, NT2-N neurons, to different concentrations of UCB and albumin at a 1.5 molar ratio and used multiple, independent measures of cell damage to evaluate neuronal injury after 6, 24, and 48 h. Low doses of UCB (0.66, 2, and 5 M) induced a moderate loss of 3-4[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) cleavage accompanied by delayed morphologic changes consistent with apoptosis (2 and 5 M). Moderate concentrations of UCB (10 and 25 M) resulted in early (6 h) necrosis in a subset of neurons, while remaining neurons underwent progressive impairment of MTT cleavage and increasing lactate dehydrogenase (LDH) release accompanied by predominantly delayed apoptosis. High concentrations of UCB (100 M) induced severe impairment of MTT cleavage, extensive LDH release, and morphologic changes consistent with necrosis within 6 h. Used as a positive control for apoptosis, 2 M STS induced progressive impairment of MTT cleavage and morphologic changes consistent with apoptosis over the entire observation period. DNA electrophoresis at 48 h was compatible with apoptosis both after treatment with STS and UCB concentrations Յ25, but not at 100 M. Cleavage of poly (ADP-ribose) polymerase (PARP) was only seen in neurons treated with low UCB concentrations and STS. We conclude that UCB induces early necrosis at high and moderate concentrations and predominantly delayed apoptosis at low and moderate concentrations in cultured human NT2-N neurons. Bilirubin-induced encephalopathy in the newborn has been described for over a century. The term is mainly used to describe a picture of reversible lethargy and alterations in cerebral evoked potentials thought to be benign (1). The term kernicterus has been used to describe more severe clinical manifestations like seizures, opisthotonos, and hyperthermia in the acutely ill newborn, followed by neurologic sequelae in survivors or death (1). The question of whether these manifestation share a common graded mechanism or reflect different entities is not settled (2).Previously, neuronal death has been thought of as the result of either necrosis or apoptosis (3). However, it now seems that apoptosis and necrosis represent extreme ends of a spectrum of possible biochemical and morphologic deaths, so that a more dynamic boundary between apoptosis and necrosis has been suggested (3). Several studies have shown that it is often the magnitude of the initial insult, rather than the specific type of the stimulus, that determines whether the cell undergoes either type of cell death (3,4). Recently, several studies have reported that UCB primarily induces apoptosis both in primary cultures of fetal cortical and cerebellar rodent neurons (5,6), in cultures

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confidence: 99%

Bilirubin Induces Apoptosis and Necrosis in Human NT2-N Neurons

et al. 2005

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“…The differentiated NT2/N neurons have been shown to co-express multiple phenotypes, including GABAergic, catecholaminergic, cholinergic, and peptidergic (38 -42). They also express pro- teins involved in exocytosis, such as synaptophysin and chromogranin (33), and form both glutamatergic excitatory and GABAergic inhibitory synapses (42,43). All these features suggest that they are able to establish functional interactions with each another and, therefore, are a convenient in vitro model of human neuronal cells.…”

Section: Resultsmentioning

confidence: 61%

Transcriptional Activation of the Human Brain-derived Neurotrophic Factor Gene Promoter III by Dopamine Signaling in NT2/N Neurons

Fang

Chartier

Sodja

et al. 2003

Journal of Biological Chemistry

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Brain-derived neurotrophic factor (BDNF) 1 is a member of neurotrophin family, structurally related to nerve growth factor, neurotrophin-3, and neurotrophin-4/5. Its biological activity is mediated by tyrosine kinase receptor B (TrkB) and its downstream signaling (1). The gene is highly expressed and widely distributed in the central nervous system, and it plays a significant role in the maintenance of function and survival of neurons (for review, see Ref.2). Evidence accumulated in recent years suggests that BDNF is also involved in the modulation of synaptic activity in the adult brain, producing long lasting changes in synaptic structure and function (for reviews, see Refs. 3-5).There is a growing interest in BDNF as a potential therapeutic agent for neurodegenerative diseases, because its deficiency was found in brains of both Alzheimer's and Parkinson's patients (6 -10). Indeed, BDNF treatment has been shown not only to potentiate synaptic transmission in vivo (11, 12) but also to increase neuronal survival and augment some behavioral changes in animal models (13,14). However, more recent data indicate that BDNF can also induce behavioral sensitization by causing an overexpression of dopamine D3 receptors and could, actually, contribute to the amplification of pathophysiologies associated with conditions such as epilepsy, drug addiction, schizophrenia, and Parkinson's disease (15, 16). Clearly, further work is required to resolve some of these potential side-effects. In contrast to a large body of work on the temporal and spatial patterns of BDNF expression in neurodevelopment and neurodegeneration, relatively little is known about the transcriptional regulation of the human BDNF gene. This is partially due to the fact that the genomic structure of the human gene has not yet been fully elucidated. The gene was first localized to chromosome11p13 and predicted to consist of multiple exons (17), but the existence of multiple transcripts, derived from different exons, was demonstrated only recently by Aoyama et al. (18) in human neuroblastoma cells. A more detailed transcript mapping of an 810-kb region of chromosome 11p13-14 further defined its genomic localization, although no additional information on the actual structure of the gene itself was presented (19).The existence of multiple human BDNF transcripts (18) is consistent with a genomic structure similar to that of the rat gene, which consists of four short 5Ј exons, each controlled by a distinct promoter, and one 3Ј exon encoding the mature BDNF protein (20,21). In the rat, the four promoters direct expression of the BDNF gene in a tissue-specific manner, i.e. promoters I and II are active preferentially in neurons, whereas promoters III and IV are active both in neurons and in a limited number of non-neuronal tissues such as lung and heart (20, 21). Thus far, only a limited characterization of a 3.2-kb genomic fragment of the human gene containing some structural elements of a promoter was reported (22).Recently, two transcription factors, CREB and ...

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“…These NTERA2-derived neurons have been extensively analyzed, and it appears that they are functionally mature, to the extent that they express a variety of neurotransmitter phenotypes (catecholinergic, cholinergic, GABAergic, and serotonergic) [Squires et al, 1996;Guillemain et al, 2000] and form functional synapses [Hartley et al, 1999].…”

Section: Principles and Advantages Of Methods Usedmentioning

confidence: 99%