• Journal of Korean Neurosurgical Society
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• 제57권5호
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• pp.335-341
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• 2015

Objective : The main causes of spinal cord ischemia are a variety of vascular pathologies causing acute arterial occlusions. We investigated neuro-protective effects of kefir on spinal cord ischemia injury in rats. Methods : Rats were divided into three groups : 1) sham operated control rats; 2) spinal cord ischemia group fed on a standard diet without kefir pretreatment; and 3) spinal cord ischemia group fed on a standard diet plus kefir. Spinal cord ischemia was performed by the infrarenal aorta cross-clamping model. The spinal cord was removed after the procedure. The biochemical and histopathological changes were observed within the samples. Functional assessment was performed for neurological deficit scores. Results : The kefir group was compared with the ischemia group, a significant decrease in malondialdehyde levels was observed (p<0.05). Catalase and superoxide dismutase levels of the kefir group were significantly higher than ischemia group (p<0.05). In histopathological samples, the kefir group is compared with ischemia group, there was a significant decrease in numbers of dead and degenerated neurons (p<0.05). In immunohistochemical staining, hipoxia-inducible factor-$1{\alpha}$ and caspase 3 immunopositive neurons were significantly decreased in kefir group compared with ischemia group (p<0.05). The neurological deficit scores of kefir group were significantly higher than ischemia group at 24 h (p<0.05). Conclusion : Our study revealed that kefir pretreatment in spinal cord ischemia/reperfusion reduced oxidative stress and neuronal degeneration as a neuroprotective agent. Ultrastructural studies are required in order for kefir to be developed as a promising therapeutic agent to be utilized for human spinal cord ischemia in the future.

• Genomics & Informatics
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• 제11권3호
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• pp.129-134
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• 2013

Orthostatic hypotension (OH) is defined by a 20-mm Hg difference of systolic blood pressure (dtSBP) and/or a 10-mm Hg difference of diastolic blood pressure (dtDBP) between supine and standing, and OH is associated with a failure of the cardiovascular reflex to maintain blood pressure on standing from a supine position. To understand the underlying genetic factors for OH traits (OH, dtSBP, and dtDBP), genome-wide association studies (GWASs) using 333,651 single nucleotide polymorphisms (SNPs) were conducted separately for two population-based cohorts, Ansung (n = 3,173) and Ansan (n = 3,255). We identified 8 SNPs (5 SNPs for dtSBP and 3 SNPs for dtDBP) that were repeatedly associated in both the Ansung and Ansan cohorts and had p-values of < $1{\times}10^{-5}$ in the meta-analysis. Unfortunately, the SNPs of the OH case control GWAS did not pass our p-value criteria. Four of 8 SNPs were located in the intergenic region of chromosome 2, and the nearest gene (CTNNA2) was located at 1 Mb of distance. CTNNA2 is a linker between cadherin adhesion receptors and the actin cytoskeleton and is essential for stabilizing dendritic spines in rodent hippocampal neurons. Although there is no report about the function in blood pressure regulation, hippocampal neurons interact primarily with the autonomic nervous system and might be related to OH. The remaining SNPs, rs7098785 of dtSBP trait and rs6892553, rs16887217, and rs4959677 of dtDBP trait were located in the PIK3AP1 intron, ACTBL2-3' flanking, STAR intron, and intergenic region, respectively, but there was no clear functional link to blood pressure regulation.

• 한국안광학회지
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• 제12권4호
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• pp.133-139
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• 2007

Calretinin의 생리적 역할은 아직 확립되지 않았지만, 칼슘 buffer로서의 작용과 칼슘 중재의 신호 변환에 적극적으로 작용하고 있다. Calretinin은 인간의 망막에서 성체 광수용체 세포와 이극세포, 수평세포에서 칼슘의 수송과 칼슘의 생리적 buffering으로서의 역할을 하고 있다. 본 연구에서 한국관 박쥐의 망막의 내핵층과 신경절세포층에서 calretinin의 면역세포화학반응 신경세포들과 그 분포 양상을 조사하여 우리는 박쥐 망막에서 calretinin에 면역세포화학반응을 일으키는 AII 무축삭세포를 내핵층에서 그리고 신경절세포층에서는 신경절세포들을 확인할 수 있었다. 이번의 관찰연구는 우리의 이전 연구와 더불어 박쥐 시력의 풀리지 않는 문제와 박쥐비행의 기동성의 독특한 행동적인 측면들을 더욱 많이 이해하는데 필요하다고 생각한다.

• Applied Microscopy
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• 제47권2호
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• pp.63-69
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• 2017

Golgi staining has been modified and developed since Camillo Golgi introduced the black reaction in 1873. This study focuses on the commonly used Golgi staining methods and presents comprehensive data regarding three Golgi staining methods along with their strong and weak points. The Golgi-Cox method uses mercuric chloride for brain tissue impregnation and is a reliable technique for analyzing the complete dendritic tree of cortical neurons. However, specimens tend to shrink during the staining steps. Recent combination of the Golgi-Cox method and immunofluorescence provides additional options for neuroscientists. Rapid Golgi staining requires osmium tetroxide for the post-fixation process. It homogenously stains whole structures of neurons and provides their detailed anatomical morphology. This staining is influenced by the age of the specimen, temperature of the laboratory, and duration of each procedure. The Golgi-Kopsch method uses formaldehyde and glutaraldehyde instead of osmium tetroxide and can be used regardless of the age of the specimen and the duration after fixation. This method is suitable for research using human brain fixed for a long time or for specimens obtained from old-aged animals. Selecting a Golgi staining protocol that is appropriate for the specimen type and research purpose is important to achieve best results.

• Korean Journal of Acupuncture
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• 제19권1호
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• pp.35-45
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• 2002

Acupuncture and herbal medicine have been used to prevent and treat the cerebrovascular accident, such as a stroke, and many studies of acupuncture and moxibustion concerning to the stroke have been undertaken in the human and various animals. However, it was not published the protective effect of the electroacupuncture (EA) of Huan-do (GB30) on the transient forebrain ischemia injury. The neuroprotective effects of EA (2 ms, 10 Hz, and 1 - 2 mA) of GB30 on the transient forebrain ischemia injury was investigated by western blot of nNOS and hematoxylin and eosin stain in Sprague-Dawley rats. The transient forebrain ischemia injury resulted in increased expression of nNOS in the brain for 6 hrs after ischemia, and EA decreased significantly expression of nNOS protein in brain increased by transient forebrain ischemia injury. The survived neuronal cell stained with hematoxylin and eosin (H&E) decreased in the hippocampus by the 7 days after ischemia comparing with the normal. Comparing with the normal, the survived neurons seriously decreased cell in the hippocampus after the injury. However, the proportion of survived neurons was increased in EA treatment. These results suggest that EA of GB30 have neuroprotective effects on transient forebrain ischemia injury.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2016년도 추계학술대회 논문집
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• pp.122.2-122.2
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• 2016

Surface topographical cues has been highlighted to control the fate of neural stem cells (NSCs). Herein we developed a hierarchically patterned substrate (HPS) platform for regulating NSC differentiation. The HPS induced cytoskeleton alignment and highly activated focal adhesion in hNSCs as indicated by enhanced expression of focal adhesion proteins such as focal adhesion kinase (FAK) and vinculin. hNSCs cultured on HPS exhibited enhanced neuronal differentiation compared to flat group. We also developed a graphene oxide (GO)-based hierarchically patterned substrates (GPS) that promote focal adhesion formation and neuronal differentiation of hNSCs. Enhanced focal adhesion and differentiation of hNSCs on the HPS was reversed by blocking the ${\beta}1$ integrin binding and mechanotransduction-associated signals including Rho-associated protein kinase (ROCK) and extracellular-regulated kinase (ERK) pathway, which may suggest a potential mechanism of beneficial effects of HPS. In addition, hNSCs on the HPS differentiated into functional neurons exhibiting sodium currents and action potentials as confirmed by whole cell patch-clamping analysis. The hierarchical topography can direct differentiation of NSCs towards functional neurons, and therefore would be an important element for the design of functional biomaterials for neural tissue regeneration applications.

• International Journal of Oral Biology
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• 제34권4호
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• pp.177-183
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• 2009

Human mesenchymal stem cell (hMSCs) isolated from human adult bone marrow have self-renewal capacity and can differentiate into multiple cell types in vitro and in vivo. A number of studies have now demonstrated that MSCs can differentiate into various neuronal populations. Due to their autologous characteristics, replacement therapy using MSCs is considered to be safe and does not involve immunological complications. The basic helix-loop-helix (bHLH) transcription factor Olig2 is necessary for the specification of both oligodendrocytes and motor neurons during vertebrate embryogenesis. To develop an efficient method for inducing neuronal differentiation from MSCs, we attempted to optimize the culture conditions and combination with Olig2 gene overexpression. We observed neuron-like morphological changes in the hMSCs under these induction conditions and examined neuronal marker expression in these cells by RTPCR and immunocytochemistry. Our data demonstrate that the combination of Olig2 overexpression and neuron-specific conditioned medium facilitates the neuronal differentiation of hMSCs in vitro. These results will advance the development of an efficient stem cell-mediated cell therapy for human neurodegenerative diseases.

• Molecular & Cellular Toxicology
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• 제4권2호
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• pp.164-169
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• 2008

Methylmercury (MeHg) is known to have devastating effects on the mammalian nervous system. In order to characterize the mechanism of MeHg-induced neurotoxicity, we investigated the analysis of transcriptional profiles on human 8k cDNA microarray by treatment of $1.4{\mu}M$ MeHg at 3, 12, 24 and 48h in human neuroblastoma SH-SY5Y cell line. Some of the identified genes by MeHg treatment were significant at early time points (3h), while that of others was at late time points (48h). The early response genes that may represent those involved directly in the MeHg response included pantothenate kinase 3, a kinase (PRKA) anchor protein (yotiao) 9, neurotrophic tyrosine kinase, receptor, type 2 gene, associated with NMDA receptor activity regulation or perturbations of central nervous system homeostasis. Also, when SH-SY5Y cells were subjected to a longer exposure (48h), a relative increase was noted in a gene, glutamine-fructose-6-phosphate transaminase 1, reported that overexpression of this gene may lead to the increased resistance to MeHg. To confirm the alteration of these genes in cultured neurons, we then applied real time-RT PCR with SYBR green. Thus, this result suggests that a neurotoxic effect of the MeHg might be ascribed that MeHg alters neuronal receptor regulation or homeostasis of neuronal cells in the early phase. However, in the late phase, it protects cells from neurotoxic effects of MeHg.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제40권4호
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• pp.173-180
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• 2014

Objectives: The purpose of this study was to investigate the neurogenic differentiation of human dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs), and stem cells from apical papilla (SCAP). Materials and Methods: After induction of neurogenic differentiation using commercial differentiation medium, expression levels of neural markers, microtubule-associated protein 2 (MAP2), class III ${\beta}$-tubulin, and glial fibrillary acidic protein (GFAP) were identified using reverse transcriptase polymerase chain reaction (PCR), real-time PCR, and immunocytochemistry. Results: The induced cells showed neuron-like morphologies, similar to axons, dendrites, and perikaryons, which are composed of neurons in DPSCs, PDLSCs, and SCAP. The mRNA levels of neuronal markers tended to increase in differentiated cells. The expression of MAP2 and ${\beta}$-tubulin III also increased at the protein level in differentiation groups, even though GFAP was not detected via immunocytochemistry. Conclusion: Human dental stem cells including DPSCs, PDLSCs, and SCAP may have neurogenic differentiation capability in vitro. The presented data support the use of human dental stem cells as a possible alternative source of stem cells for therapeutic utility in the treatment of neurological diseases.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2004년도 제47차 추계학술대회
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• pp.75-76
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• 2004