• 생명과학회지
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• 제19권8호
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• pp.1062-1066
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• 2009

N-Acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59)는 GlcNAc를 인산화하여 GlcNAc-6-phosphate를 만드는 효소이다. 효소자체에 대한 자세한 연구에도 불구하고 포유류에서NAGK의 표현에 대한 연구는 거의 없다. 배양한 흰쥐의 해마신경세포에서 NAGK은 세포체/가지돌기 영역에서 클러스터(cluster)를 형성한다. 본 연구에서는 가지돌기의 긴 축에서부터 가쪽으로 돌출되는 NAGK 클러스터에 대하여 연구하였다. 배양한 해마신경세포를 NAGK와 다양한 연접표지단백질에 대한 항체로 이중염색한 결과 NAGK 클러스터가 가지돌기의 바닥에는 있었지만 억제성 연접후부위에는 존재하지 않았다. 또한 흰쥐 전뇌(forebrain)의 균질액(homogenate, BH), 연접체(synaptosome, S), 연접후치밀질(postsynaptic density, PSD) 분획을 NAGK 항체로 면역염색한 결과 NAGK는 연접체에는 있었지만 PSD 분획에는 존재하지 않았다. 이러한 결과들은 NAGK가 가지돌기가시(spine)의 바닥쪽으로 돌출됨을 의미한다.

• Molecules and Cells
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• 제38권10호
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• pp.876-885
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• 2015

N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.

• Animal cells and systems
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• 제1권3호
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• pp.467-473
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• 1997

Taurine (2-aminoethanesulfonic acid), one of bioactive amino acid in the mammalian brain, is known to exert inhibitory effects on neurons via GABA receptor. In the present study, we examined effects of taurine on glutamateinduced neurotoxicity on hippocampal neuron cell culture using cell counting method and lactate dehydrogenase (LDH) assay. After 10 d of culture, cells were stimulated with appropriate drugs. Only 43% of cultured neuronal cells survived at one day after stimulation with 500 uM L-glutamate for 10 min. Survival rate was enhanced by 82% in the presence of 10 mM taurine. LDH activity from the culture supernatant incubated with a combination of L-glutamate and taurine was less than half of that with L-glutamate alone. In the next series of experiments, interleukin-6 (IL-6) mRNA expression in cultured astrocytes was investigated using reverse tanscription-PCR (RT-PCR). IL-6 mRNA was detected in the astrocytes stimulated with L-glutamate in a dose-dependent manner, while not detected in the unstimulated control astrocytes. The expression of IL-6 mRNA caused by 10 mM glutamate was inhibited by taurine, but not by GABA. These findings demonstrated a neuroprotective action of taurine against glutamate-induced toxicity.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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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.

• Journal of Microbiology and Biotechnology
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• 제28권1호
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• pp.12-21
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• 2018

Photorhabdus temperata (PT), a gram-negative bacterium, lives symbiotically within entomopathogenic nematodes. The insecticidal compounds derived from Photorhabdus are used as biopesticides in agriculture. However, the physiological properties are not well characterized. In the course of our screening for neuroprotective and anti-neuroinflammatory substances from natural products, the culture broth of PT showed considerable activities. By activity-guided purification, five anthraquinones, namely, 3-methoxychrysazine (1), 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone (2), 1,3,8-trihydroxy-9,10-anthraquinone (3), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (4), and 1,3,4-trimethoxy-8-hydroxy-9,10-anthraquinone (5), were isolated from the ethyl acetate fraction of the PT culture broth. Among the isolated compounds, $75{\mu}M$ 3 significantly protected mouse hippocampal neuronal cells (HT22) against 5 mM glutamate-induced cell death via the inhibition of reactive oxygen species production, $Ca^{2+}$ influx, and lipid peroxidation. Additionally, 3 and 4 effectively suppressed the interferon-${\gamma}$-induced neuroinflammation of mouse-derived microglial cells (BV2) at 10 ng/ml, via the reduction of nitric oxide, interleukin-6, and tumor necrosis factor-${\alpha}$. Anthraquinones 3 and 4 derived from the PT culture broth are a potential starting point to discover neuroprotective and anti-neuroinflammatory drug leads. The novel compound 5 is reported for the first time in this study.

• 동의신경정신과학회지
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• 제20권2호
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• pp.1-17
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• 2009

Objectives : Alzheimer's disease(AD) is the most common form of dementia, which is characterized by progressive deterioration of memory and higher cortical functions that ultimately results in total degradation of intellectual and mental activities. Nokyongdaebo-tang(Lurongdabutang) has been usually used for the treatment for the deficiency syndrome dementia and amnesia. This experiment was designed to investigate the effect of the Nokyongdaebo-tang(Lurongdabutang) hot water extract on pathological AD model. Methods : The effects of the Nokyongdaebo-tang(Lurongdabutang) hot water extract on cultured spinal cord cells induced by ${\beta}$-amyloid were investigated. The effects of the Nokyongdaebo-tan(Lurongdabutang) hot water extract on the memory deficit mice induced by scopolamine were investigated. Results : 1. ${\beta}$-amyloid treatment on cultured spinal cord cells increased both GFAP-staining intensity of astrocytes and caspase 3 immunoreactivity on cultured cells. Then, Nokyongdaebo-tang(Lurongdabutang) treatment reduced the labeling intensity for both GFAP and caspase 3 proteins in culture cells. 2. Scopolamine treatment into mice increased levels of GFAP-positive astrocytes and caspase 3-labeled cells of the hippocampal subfields dentate hilar region, CA3 and CA1 area. In vivo administration of Nokyongdaebo-tang(Lurongdabutang) attenuated labeling intensity for those two proteins in the same hippocampal areas. Similar effects were observed by the treatment of galanthamine, an inhibitor of acetylcholinesterase. Conclusions : This experiment shows that the Nokyongdaebo-tang(Lurongdabutang) may play a protective role in damaged neural tissues. Since neuronal damage seen in degenerative brains such as AD are largely unknown, the current data may provide possible insight into therapeutic strategies for AD treatments. Nokyongdaebo-tang(Lurongdabutang) might be effective for the prevention and treatment of AD.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.278.1-278.1
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• 2013

Among the prerequisites for stable neural interfacing are the long-term stability of electrical performance of and the excellent biocompatibility of conducting materials in implantable neural electrodes. Reduced graphene oxide offers a great potential for a variety of biomedical applications including biosensors and, particularly, neural interfaces due to its superb material properties such as high electrical conductivity, decent optical transparency, facile processibility, and etc. Nonetheless, there have been few systematic studies on the graphene-based neural interfaces in terms of biocompatibility of electrode materials and long term stability in electrical characteristics. In this research, we prepared the primary culture of rat hippocampal neurons directly on reduced graphene oxide films which is chosen as a model electrode material for the neural electrode. We observed that the viability of primary neuronal culture on the present structure is minimally affected by nanoscale graphene flakes below. These results implicate that the multilayer films of reduced graphene oxides can be utilized for the next-generation neural interfaces with decent biocompatibility and outstanding electrical performance.

• 한국식생활문화학회지
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• 제36권6호
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• pp.633-639
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• 2021

Raphanus sativus var. hortensis f. raphanistroides Makino (Korean wild radish [WR]) are root vegetables belonging to the Brassicaceae family. These radish species mostly grow in sea areas in Asia, where they have been traditionally used as a medicinal food to treat various diseases. To investigate the effect of WR on neuronal cell death in SH-SY5Y cells, beta-amyloid was used to develop the cell death model. WR attenuated neuronal cell death in SH-SY5Y and regulated the mitogen-activated protein kinase (MAPK) signaling. WR extract also inhibited acetylcholinesterase inhibitor activity. Additionally, the WR treatment group ameliorated the behavior of the memory-impaired mice in a scopolamine-induced mouse model. In the behavior test, WR treated mice showed shorter escape latency and swimming distance and improved the platform-crossing number and the swimming time within the target quadrant. Furthermore, WR prevented histological loss of neurons in hippocampal CA1 regions induced by scopolamine. This study shows that WR can prevent memory impairment which may be a crucial way for the prevention and treatment of memory dysfunction and neuronal cell death.

• 동의생리병리학회지
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• 제19권2호
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• pp.416-425
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• 2005

This study demonstrated neuroprotective and anti-oxidative effects of Puerariae Radix for cerebral ischemia. Neuroprotective effects were studied by using oxygen/glucous deprivation of the organotypic hippocampal slice cultures to complement limitations of in vivo and in vitro models for cerebral ischemia study. Anti-oxidative effects were studied on BV-2 microglia cells damaged by $H_2O_2$ and nitric oxide. The results obtained are as follows; The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in CA1 region of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in DG region of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of TUNEL-positive cells in both CA1 region and DG region of ischemic damaged hippocampus cultures. The group treated with $50\;{\mu}g/m{\ell}$ of Puerariae Radix demonstrated significant decrease of TUNEL-positive cells in CA1 region. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of LDH concentrations in culture media of ischemic damaged hippocampus cultures. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant increases of cell viabilities of BV-2 microglia cells damaged by $H_2O_2$. The group treated with $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant increase of cell viability of BV-2 microglia cells damaged by nitric oxide. These results suggested that Puerariae Radix of cerebral ischemic revealed neuroprotective effects through the control effect of apoptosis and oxidative damages.

• 생명과학회지
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• 제17권12호
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• pp.1627-1633
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• 2007

c-Jun N-terminal kinase (JNK)-interacting protein 1(JIP1)은 비계단백질(scaffold protein)로서 신경 세포와 ??장${\beta}$세포에서 많이 발현된다. 본 연구에서는 배양한 흰쥐 해마신경세포에서 JIP1, JIP-2 및 JIP-3을 모두 인식하는 항체를 이용하여 이들의 세포내 표현을 조사하였다. 전반적으로 JIP은 세포체와 가지돌기에 반점 모양으로 표현되었다. 이 JIP 반점들을 통계적으로 분석한 결과 흥분성 연접후표지인 PSD95 및 ${\alpha}CaMKII$ 반점 과 각각 $54.8{\pm}4.0%$ 및 $94.1{\pm}4.5%$가 겹쳐졌다. 반면에 억제성 연접후표지인 그리신수용체 및 gephyrin 반점과는 각각 단지 $8.6{\pm}0.5%$ 및 $7.3{\pm}0.5%$만 겹쳐졌다. 한편 lipid raft의 표지인 flotillin 반점의 상당부분$(29.3{\pm}1.0%)$이 JIP 반점과 겹쳐졌다. 또한, JIP은 일부 가지돌기의 끝부분에 매우 강하게 발편되었으며 축삭에는 표현이 미미하였다. 이 결과들은 JIP 단백질이 흥분성 연접후구역, 일부 lipid raft, 그리고 일부 가지돌기 끝부분에 주로 위치함을 의미한다.