• Clinical and Experimental Pediatrics
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• 제45권6호
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• pp.732-742
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• 2002

목 적: 저산소-허혈에 대한 신경독성의 규명 및 xanthine oxidase inhibitor인 allopurinol의 저산소성-허혈 유도에 미치는 방어효과를 조사하기 위하여 본 연구를 시도하였다. 방 법: 신생쥐에 우측 총경동맥을 결찰 및 8% O2의 노출로 허혈 및 저산소 상태를 만든 후 저산소성-허혈이 12-72시간 동안 대뇌의 neuron과 astrocyte에 미치는 영향을 조사하여 신경독성을 규명하고, 또한 xanthine oxidase inhibitor인 allopurinol이 저산소성-허혈 유도에 미치는 영향을 조사하기 위하여 저산소성-허혈 유도 15분 전에 150 mg/kg의 allopurinol을 복강 투여한 다음 투여 후 14일 후에 신생쥐를 희생하여 이의 뇌 조직으로부터 순수분리 배양한 신경 세포에 대하여 세포의 수적 변화와 생존율을 비롯하여 LDH와 단백질합성 및 PKC를 조사하였다. 결 과 : 1) 저산소성-허혈은 저산소-허혈 유도 직후부터 72시간 동안 시간경과에 비례하여 신생쥐의 대뇌 neurons의 수와 세포생존율을 유의하게 감소시켰다. 2) 저산소성-허혈은 저산소-허혈 유도 직후부터 72시간 동안 시간경과에 비례하여 신생쥐의 대뇌 astrocyte의수와 세포생존율을 다소 감소시켰다. 3) 저산소-허혈 유도 14일 후 neuron의 수와 세포 생존율 및 단백질합성은 대조군에 비하여 유의하게 감소하였으나 LDH는 매우 증가하였다. 4) 저산소-허혈 유도 직전 allopurinol 처리에 의하여 neuron의 수와 세포생존율 및 단백질합성은 유의하게 증가하였고 LDH치는 현저히 감소하였다. 5) 배양된 neuron에 대한 PKC 조사에 있어서 허혈 유도 10분에 현저한 PKC치의 증가를 보였으며, allopurinol의 전 처리는 허혈 유도에 의한 PKC치의 증가를 유의하게 감소시켰다. 결 론 : 저산소성-허혈은 신생쥐의 대뇌 신경세포에 독성효과를 나타냈으며 활성산소 제거제인 allopurinol은 세포수 및 세포생존률의 증가에 의한 신경세포의 손상보호를 나타내었고, 단백질합성 증가, LDH치 및 PKC치의 감소로 세포손상에 대한 효과적 방어도 관찰할 수 있었다.

• International Journal of Stem Cells
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• 제16권3호
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• pp.269-280
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• 2023

Background and Objectives: The colonic epithelial layer is a complex structure consisting of multiple cell types that regulate various aspects of colonic physiology, yet the mechanisms underlying epithelial cell differentiation during development remain unclear. Organoids have emerged as a promising model for investigating organogenesis, but achieving organ-like cell configurations within colonic organoids is challenging. Here, we investigated the biological significance of peripheral neurons in the formation of colonic organoids. Methods and Results: Colonic organoids were co-cultured with human embryonic stem cell (hESC)-derived peripheral neurons, resulting in the morphological maturation of columnar epithelial cells, as well as the presence of enterochromaffin cells. Substance P released from immature peripheral neurons played a critical role in the development of colonic epithelial cells. These findings highlight the vital role of inter-organ interactions in organoid development and provide insights into colonic epithelial cell differentiation mechanisms. Conclusions: Our results suggest that the peripheral nervous system may have a significant role in the development of colonic epithelial cells, which could have important implications for future studies of organogenesis and disease modeling.

• Applied Microscopy
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• 제28권2호
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• pp.139-158
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• 1998

The development of the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm of crown-rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cell, primitive neuroblast, primitive supporting cell, and unmyelinated fibers. At 70mm fetus, the neuroblasts and their processes were ensheated by the bodies or processes of satellite cells. The cytoplasm of the neuroblast contained rough endoplasmic reticulum, mitochondria, Golgi complex, Nissl bodies and dense-cored vesicles. As the neuroblasts grew and differentiated dense-cored vesicles moved away from perikaryal cytoplasm into developing processes. Synaptic contacts between the cholinergic axon and dendrites of postganglionic neuron and a few axosomatic synapses were first observed at 70 mm fetus. At 90 mm fetus the superior cervical ganglion consisted of neuroblasts, satellite cells, granule-containing cells, and unmyelinated nerve fibers. The ganglion cells increased somewhat in numbers and size by 150 mm fetus. Further differentiation resulted in the formation of young ganglion cells, whose cytoplasm was densely filled with cell organelles. During next prenatal stage up to 260 mm fetus, the cytoplasm of the ganglion cells contained except for large pigment granules, all intracytoplasmic structures which were also found in mature superior cervical ganglion. A great number of synaptic contact zones between the cholinergic preganglionic axon and the dendrites of the postganglionic neuron were observed and a few axosomatic synapses were also observed. Two morphological types of the granule-containing cells in the superior cervical ganglion were first identified at 90 mm fetus. Type I granule-containing cell occurred in solitary, whereas type II tended to appeared in clusters near the blood capillaries. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma of type I granule-containing cells and preganglionic axon termials were observed. In addition, synaptic junctions between the processes of the granule- containing cells and dendrites of postganglionic neuron were also observed from 150 mm fetus onward. In conclusion, superior cervical ganglion cells and granule-containing cells arise from a common undifferentiated cell precursor of neural crest . The granule-containg cells exhibit a local modulatory feedback system in the superior cervical ganglion and nay serve as interneurons between the preganglionic and postganglionic cells.

• Interdisciplinary Bio Central
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• 제4권3호
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• pp.6.1-6.12
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• 2012

Parkinson's disease (PD) is a complicated neurodegenerative disorder although it is oftentimes defined by clinical motor symptoms originated from age dependent and progressive loss of dopaminergic neurons in the midbrain. The pathogenesis of PD involves dopaminergic and nondopaminergic neurons in many brain regions and the molecular mechanisms underlying the death of different cell types still remain to be elucidated. There are indications that PD causing disease processes occur in a global scale ranging from DNA to RNA, and proteins. Several PD-associated genes have been reported to play diverse roles in controlling cellular functions in different levels, such as chromatin structure, transcription, processing of mRNA, translational modulation, and posttranslational modification of proteins. The advent of quantitative high throughput screening (HTS) tools makes it possible to monitor systemic changes in DNA, RNA and proteins in PD models. Combined with dopamine neuron isolation or derivation of dopamine neurons from PD patient specific induced pluripotent stem cells (PD iPSCs), HTS techonologies will provide opportunities to draw PD causing sequences of molecular events in pathologically relevant PD samples. Here I discuss previous studies that identified molecular functions in which PD genes are involved, especially those signaling pathways that can be efficiently studied using HTS methodologies. Brief descriptions of quantitative and systemic tools looking at DNA, RNA and proteins will be followed. Finally, I will emphasize the use and potential benefits of PD iPSCs-derived dopaminergic neurons to screen signaling pathways that are initiated by PD linked gene mutations and thus causative for dopaminergic neurodegneration in PD.

• 대한한의학방제학회지
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• 제8권1호
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• pp.319-328
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• 2000

To evaluate the effect of Baepungtang(BPT) water extract on cultured mouse spinal sensory neuron which was inhibited by xanthine oxidase(XO) and hypoxanthine(HX)-induced oxigen radicals, MTT assay, NR assay, Neurofilament enzymeimmuno assay and LDH activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of BPT water extract for 3 hours prior to exposure of XO/HX. The results obtained were as follows: 1. XO/HX, a oxigen radical, decreased the survival rate of the cultured mouse spinal sensory neuron cell on NR assay and MTT assay. 2. $MTT_{50}$ value and $NR_{50}$ value of XO/HX were 30 mU/ml XO/O.2 mM HX. 3. BPT water extract have efficacy of increasing neurofilament. 4. BPT water extract have efficacy of increasing LDH activity. From above the results, It is concluded that BPT has marked efficacy as a treatment for the damages caused in the XO/HX-mediated oxidative process.

• 전자공학회논문지S
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• 제35S권11호
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• pp.88-95
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• 1998

본 논문은 인공생명 기법을 이용하여 생물의 정보처리 시스템을 구현하고자 하는 것이다. 자연계의 생물은 그 자체로 훌륭한 정보처리 시스템이다. 생물체는 하나의 생식 세포로부터 발생된다. 또한 이 개체의 종은 진화의 과정을 통해 환경에 적응한다. 본 논문에서는 이와 같은 생물학적인 발생과 진화의 개념을 이용하여 신경망을 설계하는 방법을 제안한다. 생물체의 개체발생은 발생모델의 하나인 셀룰라 오토마다(CA)를 통하여 구현하였고 진화과정은 진화 알고리즘(EAs)을 사용하였다. 우리는 이와 같이 구현한 '진화하는 셀룰라 오토마타 신경망'을 줄여서 ECANS1이라 명명하였다. 셀 사이의 연결은 CA 법칙에 의하여 결정되며, 셀의 초기 패턴이 진화함으로써 유용한 신경망을 찾아낸다. 신경망의 각 셀 즉 뉴런은 생물의 발화 ${\cdot}$ 비발화의 특성을 갖는 카오스 뉴런 모델을 사용하였다. 그리고 신경마의 최종 출력값은 뉴런의 발화 빈도로서 나타내었다. 제안한 방법은 Exclusive-OR 문제 및 패리티 문제에 적용함으로써 그 유효성을 검증하였다.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2005년도 학술대회 논문집 정보 및 제어부문
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• pp.38-40
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• 2005

Microdrive with high precision and light mass enough to install on mouse head was fabricated for recording the reliable signal of neuron cell to understand the brain study. The proposed microdrive has three H-form PZT actuators and its guide structure. The microdrive operation principle is based on the well known inchworm principle. The synchronization of three PZT actuators is able to produce the linear motion along the guide structure. Our proposed microdrive has a precise accuracy of about 100nm and a long stroke of about 5mm. The electrode which is used for the recording of the action potential of the neuron cell was fixed at one of PZT actuators. The proposed microdrive was suited to acquisition of signals from in vivo extra-cellular single-unit recoding. On the condition of the anesthetized mouse, the single-unit signals could be recorded by using the proposed microdrive. In addition, applying the PZT microdrive to an alert mouse, we try to implant it on a mouse brain skull to explore single neuron firing.

• International Journal of Oral Biology
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• 제33권4호
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• pp.217-221
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• 2008

Bicuculline is one of the most commonly used $GABA_A$ receptor antagonists in electrophysiological research. Because of its poor water solubility, bicuculline quaternary ammonium salts such as bicuculline methiodide (BMI) and bicuculline methbromide are preferred. However, a number of studies have shown that BMI has non-$GABA_A$ receptor-mediated effects. The substantia gelatinosa (SG) of the trigeminal subnucleus caudalis (Vc) is implicated in the processing of nociceptive signaling. In this study, we investigated whether BMI has non-GABA receptor-mediated activity in Vc SG neurons using a whole cell patch clamp technique. SG neurons were depolarized by application of BMI ($20{\mu}M$) using a high $Cl^-$ pipette solution. GABA ($30-100{\mu}M$) also induced membrane depolarization of SG neuron. Although BMI is known to be a $GABA_A$ receptor antagonist, GABA-induced membrane depolarization was enhanced by co-application with BMI. However, free base bicuculline (fBIC) and picrotoxin (PTX), a $GABA_A$ and $GABA_C$ receptor antagonist, blocked the GABA-induced response. Furthermore, BMI-induced membrane depolarization persisted in the presence of PTX or an antagonist cocktail consisting of tetrodotoxin ($Na^+$ channel blocker), AP-5 (NMDA receptor antagonist), CNQX (non-NMDA receptor antagonist), and strychnine (glycine receptor antagonist). Thus BMI induces membrane depolarization by directly acting on postsynaptic Vc SG neurons in a manner which is independent of $GABA_A$ receptors. These results suggest that other unknown mechanisms may be involved in BMI-induced membrane depolarization.

• 동의생리병리학회지
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• 제19권5호
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• pp.1303-1310
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• 2005

This study was performed to investigate the effect of Bunsimgieumgagam on the stress due to the maternal separation in rat. In this study, we researched in 'the behavioral observation', 'the changes of body weight', 'quantitative analysis of the number of BrdU-positive cells per section in dentate gyrus of hippocampus', 'free radical scavenging assay' and 'MTT-based cytotoxicity assay of SK-N-SH cell line', in order to figure out the effect on which Bunsimgieumgagam has the increase of neuron in dentate gyrus of hippocampus damaged by the stress due to the maternal separation. In the behavioral Observation, Bunsimgieumgagam was also efficacious against the decline of one's behavior and anorexia derived from the stress by the maternal separation. In the change of body weight, it showed that the Bunsimgieumgagam is effective in the recovery of weight loss caused by heavy stress(p<0.05). Also, Bunsimgieumgagam had an increasing effect, which is similar to a normal state, on DG's neuron in hippocampus (P<0.001). In free radical scavenging assay, Bunsimgieumgagam had a superior free radical scavenging effect. And it showed a significant result with the high cell proliferation effect in MTT-based cytotoxicity assay(P<0.01, p<0.001) This result suggest that Bunsimgieumgagam has an anti-stress effect and a proliferation effect of neuron in dentate gyrus of hippocampus, and it shows the potential of Bunsimgieumgagam in the treatment for the various disorders derived from children's stress.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2008년도 제49회 학술심포지움 및 국제학술대회
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• pp.63.2-63.2
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• 2008