• Title/Summary/Keyword: neuronal

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Characterization of Dopaminergic Neuronal Cell Death Induced by either N-Methyl-4-Phenylpyridinium of 6-hydroxydopamine (N-메칠-4-페닐피리디니움 및 6-히드록시도파민으로 유도된 도파민계 신경세포 사멸 기작의 규명)

  • O, Yeong-Jun;Choi, Won-Seok
    • YAKHAK HOEJI
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    • v.41 no.1
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    • pp.86-93
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    • 1997
  • Even though both N-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP) and 6-hydroxydopamine have been widely used to establish the experimental model for dopaminergic neuronal ce ll death. mechanisms underlying this phenomenon have not been firmly explored. To investigate how these dopaminergic neurotoxins induce neuronal cell death, murine dopaminergic neuronal cell line, MN9D cells were treated with various concentration of either 6-hydroxydopamine or active form of MPTP, N methyl-4-phenylpyridinium (MPP$^+$). Treatment of cells with 5-100 uM 6-hydroxydopamine resulted in apoptotic cell death whereas cell death induced by 5~50 uM MPP$^+$ was not demonstrated typical apoptotic characteristics such as cell shrinkage, apoptotic body and nuclear condensation. Cell death induced by 6-hydroxydopamine was partially blocked in the presence of antioxidants including soluble form of vitamin E or desferrioxamine suggesting that generation of oxidative stress may be associated with 6-hydroxydopamine-induced cell death in MN9D cells. In contrast, MPP$^+$-induced cell death was not blocked by treatment with any of antioxidants tested. As previously demonstrated that MPP$^+$ caused metabolic alterations such as glucose metabolism, removal of glucose from the medium partially inhibited MPP$^+$-induced cell death suggesting excessive cycles of glycolysis may be associated with MPP$^+$-induced cell death. Taken together, these studies demonstrate that two types of dopaminergic neurotoxins recruit distinct neuronal cell death pathways.

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Preventive effects of imperatorin on perfluorohexanesulfonate-induced neuronal apoptosis via inhibition of intracellular calcium-mediated ERK pathway

  • Lee, Eunkyung;Choi, So-Young;Yang, Jae-Ho;Lee, Youn Ju
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.4
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    • pp.399-406
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    • 2016
  • Early life neuronal exposure to environmental toxicants has been suggested to be an important etiology of neurodegenerative disease development. Perfluorohexanesulfonate (PFHxS), one of the major perfluoroalkyl compounds, is widely distributed environmental contaminants. We have reported that PFHxS induces neuronal apoptosis via ERK-mediated pathway. Imperatorin is a furanocoumarin found in various edible plants and has a wide range of pharmacological effects including neuroprotection. In this study, the effects of imperatorin on PFHxS-induced neuronal apoptosis and the underlying mechanisms are examined using cerebellar granule cells (CGC). CGC were isolated from seven-day old rats and were grown in culture for seven days. Caspase-3 activity and TUNEL staining were used to determine neuronal apoptosis. PFHxS-induced apoptosis of CGC was significantly reduced by imperatorin and PD98059, an ERK pathway inhibitor. PFHxS induced a persistent increase in intracellular calcium, which was significantly blocked by imperatorin, NMDA receptor antagonist, MK801 and the L-type voltage-dependent calcium channel blockers, diltiazem and nifedipine. The activation of caspase-3 by PFHxS was also inhibited by MK801, diltiazem and nifedipine. PFHxS-increased ERK activation was inhibited by imperatorin, MK801, diltiazem and nifedipine. Taken together, imperatorin protects CGC against PFHxS-induced apoptosis via inhibition of NMDA receptor/intracellular calcium-mediated ERK pathway.

Neuroprotective effects of the antioxidant action of 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride against ischemic neuronal damage in the brain

  • Ha, Seung Cheol;Han, A Reum;Kim, Dae Won;Kim, Eun-A;Kim, Duk-Soo;Choi, Soo Young;Cho, Sung-Woo
    • BMB Reports
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    • v.46 no.7
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    • pp.370-375
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    • 2013
  • Ischemia is characterized by oxidative stress and changes in the antioxidant defense system. Our recent in vitro study showed that 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride protects cortical astrocytes against oxidative stress. In the current study, we examined the effects of 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride on ischemia-induced neuronal damage in a gerbil ischemia/reperfusion models. Extensive neuronal death in the hippocampal CA1 area was observed 4 days after ischemia/reperfusion. Intraperitoneal injection of 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride (0.3 mg/kg body weight) significantly prevented neuronal death in the CA1 region of the hippocampus in response to transient forebrain ischemia. 2-Cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride administration reduced ischemia-induced increases in reactive oxygen species levels and malondialdehyde content. It also attenuated the associated reductions in glutathione level and superoxide dismutase, catalase, and glutathione peroxidase activities. Taken together, our results suggest that 2-cyclopropylimino-3-methyl-1,3-thiazoline hydrochloride protects against ischemia-induced neuronal damage by reducing oxidative stress through its antioxidant actions.

Inhibition of Oxidative Stress-induced and Excitotoxic Neuronal Cell Damage by Xuesaitong Ruanjiaonang (혈색통연교낭(血塞通軟膠囊)의 산화적 및 흥분성 신경세포독성 억제작용)

  • Cho Jungsook
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.30-37
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    • 2005
  • Xuesaitong Ruanjiaonang (XR), a soft capsule containing Panax notoginseng saponins as main ingredients, is believed to remove extravasated blood and increase cerebral blood flow by improving blood circulation, and therefore, has been used in China to treat ischemic stroke or hemiplegia caused by cerebral thrombosis. To characterize pharmacological actions of XR, the present study evaluated its effects on neuronal cell damage induced by various oxidative insults or excitotoxic amino acids in primary cultured rat cortical cells. The neuronal cell viability was not affected by XR with the exposure for 2 h at the concentrations tested in this study ($10{\sim}1000\;{\mu}g/ml$). However, significant reduction of the cell viability was observed when the cultured cells were exposed to XR at $1000\;{\mu}g/ml$ for 24 h. XR was found to concentration-dependently inhibit the oxidative neuronal damage induced by $H_{2}O_2$, xanthine/xanthine oxidase or $Fe^{2+}$/ascorbic acid. In addition, it dramatically inhibited the excitotoxic damage induced by glutamate or N-methyl-D-aspartate (NMDA). We found that the NMDA-induced neurotoxicity was inhibited more effectively and potently than the glutamate-induced toxicity. Moreover, XR was found to exert mild inhibition of lipid peroxidation induced by $Fe^{2+}$/ascorbic acid in rat brain homogenates and some 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together, these results demonstrate neuroprotective and antioxidant effects of XR, showing inhibition of oxidative and excitotoxic damage in the cultured cortical neurons, as well as inhibition of lipid peroxidation and its radical scavenging activity. Considering that excitotoxicity and oxidative stress pl ay crucial roles in neuronal cell damage during ischemia and reperfusion, these results may provide pharmacological basis for its clinical usage to treat ischemic stroke.

Generation of Neural Progenitor Cells from Pig Embryonic Germ Cells

  • Choi, Kwang-Hwan;Lee, Dong-Kyung;Oh, Jong-Nam;Kim, Seung-Hun;Lee, Mingyun;Jeong, Jinsol;Choe, Gyung Cheol;Lee, Chang-Kyu
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.42-49
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    • 2020
  • As a preclinical study, many researchers have been attempted to convert the porcine PSCs into several differentiated cells with transplantation of the differentiated cells into the pigs. Here, we attempted to derive neuronal progenitor cells from pig embryonic germ cells (EGCs). As a result, neuronal progenitor cells could be derived directly from pig embryonic germ cells through the serum-free floating culture of EB-like aggregates (SFEB) method. Treating retinoic acid was more efficient for inducing neuronal lineages from EGCs rather than inhibiting SMAD signaling. The differentiated cells expressed neuronal markers such as PAX6, NESTIN, and SOX1 as determined by qRT-PCR and immunostaining. These data indicated that pig EGCs could provide valid models for human therapy. Finally, it is suggested that developing transgenic pig for disease models as well as differentiation methods will provide basic preclinical data for human regenerative medicine and lead to the success of stem cell therapy.

Effects of Glutamate Receptor Antagonists and Protein Synthesis Inhibitor on Delayed Neuronal Death Induced by Transient Global Ischemia in Rat Brain

  • Ko, Jun-Seog;Bae, Choon-Sang;Kim, Jong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.3
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    • pp.279-286
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    • 1998
  • It has been well documented that transient forebrain global ischemia causes selective neuronal degeneration in hippocampal CA1 pyramidal neurons with a delay of a few days. The mechanism of this delayed hippocampal CA1 pyramidal neuronal death (DND) is still controversial. To delineate the mechanisms of the DND, the effects of treatment with MK-801, an NMDA receptor antagonist, kynurenic acid, a NMDA/non-NMDA receptor antagonist, and/or cycloheximide, a protein synthesis inhibitor, on the DND were investigated in male Wistar rats. To examine the participation of apoptotic neuronal death in the DND, TUNEL staining was performed in ischemic brain section. Global ischemia was induced by 4-vessel occlusion for 20 min. All animals in this study showed the DND 3 and 7 days after the ischemic insult. The DND that occured 3 days and 7 days after the ischemia were not affected by pretreatment with MK-801 (1 mg/kg), but markedly attenuated by the pretreatment with kynurenic acid (500 mg/kg). Treatment with cycloheximide (1 mg/kg) also markedly inhibited the DND. The magnitudes of attenuation by the two drugs were similar. The magnitude of attenuation by co-treatments with kynurenic acid and cycloheximide was not greater than that with any single treatment. TUNEL staining was negative in the sections obtained 1 or 2 days after the ischemic insults, but it was positive at hippocampal CA1 pyramidal cells in sections collected 3 days after the ischemia. These results suggested that the DND should be mediated by the activation of non-NMDA receptor, not by the activation of NMDA receptor and that the activation of AMPA receptor should induce the apoptotic process in the DND.

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Investigating the Morphology and Kinetics of Three-Dimensional Neuronal Networks on Electro-Spun Microstructured Scaffolds

  • Kim, Dongyoon;Kim, Seong-Min;Kang, Donghee;Baek, Goeun;Yoon, Myung-Han
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.277.2-277.2
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    • 2013
  • Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

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Neurogenesis and neuronal migration of dopaminergic neurons during mesencephalon development in mice

  • Kim, Mun-ki;Lee, Si-Joon;Vasudevan, Anju;Won, Chungkil
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.125-129
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    • 2018
  • Dopaminergic neurons are one of the major neuronal components in the brain. Mesencephalon dopamine (DA) neurogenesis takes place in the ventricular zone of the floor plate, when DA progenitors divide to generate postmitotic cells. These cells migrate through the intermediate zone while they differentiate and become DA neurons on reaching the mantle zone. However, neurogenesis and neuronal migration on dopaminergic neurons remain largely unexplored in the mesencephalon development. This study presents neurogenesis and neuronal migration patterns of dopaminergic neurons during mesencephalic development of the mouse. Neurons from embryonic day (E) 10-14 were labelled by a single injection of 5-bromodeoxyuridine and immunohistochemistry was performed. The neurogenesis occurred mainly at the E10 and E11, which was uniformly distributed in the mesencephalic region, but neurons after E13 were observed only in the dorsal mesencephalon. At the postnatal day 0 (P0), E10 generated neurons were spread out uniformly in the whole mesencephalon whereas E11-originated neurons were clearly depleted in the red nucleus region. DA neurons mainly originated in the ventromedial mesencephalon at the early embryonic stage especially E10 to E11. DA neurons after E12 were only observed in the ventral mesencephalon. At E17, E10 labelled neurons were only observed in the substantia nigra (SN) region. Our study demonstrated that major neurogenesis occurred at E10 and E11. However, neuronal migration continued until neonatal period during mesencephalic development.

Alpha-lipoic acid protects human dopaminergic neuronal cells against hydrogen peroxide-induced cell injury by inhibiting autophagy and apoptosis

  • Kang, Kyeong-Rok;Kim, Jae-Sung;Kim, Tae-Hyeon;Seo, Jeong-Yeon;Lim, HyangI;Park, Jong-Hyun;Yang, Kwang Yeol;Yu, Sun-Kyoung;Kim, Heung-Joong;Kim, Chun Sung;Chun, Hong Sung;Lee, Dong-Seol;Park, Joo-Cheol;Kim, Do Kyung
    • International Journal of Oral Biology
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    • v.46 no.1
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    • pp.15-22
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    • 2021
  • Alpha-lipoic acid (ALA) is a naturally occurring antioxidant and has been previously used to treat diabetes and cardiovascular disease. However, the autophagy effects of ALA against oxidative stress-induced dopaminergic neuronal cell injury remain unclear. The aim of this study was to investigate the role of ALA in autophagy and apoptosis against oxidative stress in the SH-SY5Y human dopaminergic neuronal cell line. We examined SH-SY5Y phenotypes using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay (cell viability/proliferation), 4′,6-diamidino-2-phenylindole dihydrochloride nuclear staining, Live/Dead cell assay, cellular reactive oxygen species (ROS) assay, immunoblotting, and immunocytochemistry. Our data showed ALA attenuated hydrogen peroxide (H2O2)-induced ROS generation and cell death. ALA effectively suppressed Bax up-regulation and Bcl-2 and Bcl-xL down-regulation. Furthermore, ALA increased the expression of the antioxidant enzyme, heme oxygenase-1. Moreover, the expression of Beclin-1 and LC-3 autophagy biomarkers was decreased by ALA in our cell model. Combined, these data suggest ALA protects human dopaminergic neuronal cells against H2O2-induced cell injury by inhibiting autophagy and apoptosis.

The CCAAT-box transcription factor, NF-Y complex, mediates the specification of the IL1 neurons in C. elegans

  • Woojung Heo;Hyeonjeong Hwang;Jimin Kim;Seung Hee Oh;Youngseok Yu;Jae-Hyung Lee;Kyuhyung Kim
    • BMB Reports
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    • v.56 no.3
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    • pp.153-159
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    • 2023
  • Neuronal differentiation is highly coordinated through a cascade of gene expression, mediated via interactions between trans-acting transcription factors and cis-regulatory elements of their target genes. However, the mechanisms of transcriptional regulation that determine neuronal cell-fate are not fully understood. Here, we show that the nuclear transcription factor Y (NF-Y) subunit, NFYA-1, is necessary and sufficient to express the flp-3 neuropeptide gene in the IL1 neurons of C. elegans. flp-3 expression is decreased in dorsal and lateral, but not ventral IL1s of nfya-1 mutants. The expression of another terminally differentiated gene, eat-4 vesicular glutamate transporter, is abolished, whereas the unc-8 DEG/ENaC gene and pan-neuronal genes are expressed normally in IL1s of nfya-1 mutants. nfya-1 is expressed in and acts in IL1s to regulate flp-3 and eat-4 expression. Ectopic expression of NFYA-1 drives the expression of flp-3 gene in other cell-types. Promoter analysis of IL1-expressed genes results in the identification of several cis-regulatory motifs which are necessary for IL1 expression, including a putative CCAAT-box located in the flp-3 promoter that NFYA-1 directly interacts with. NFYA-1 and NFYA-2, together with NFYB-1 and NFYC-1, exhibit partly or fully redundant roles in the regulation of flp-3 or unc-8 expression, respectively. Taken together, our data indicate that the NF-Y complex regulates neuronal subtype-specification via regulating a set of terminal-differentiation genes.