• 운동과학
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• 제26권3호
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• pp.229-237
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• 2017

PURPOSE: This study was to investigate the Glycolysis mediated sarcoplasmic reticulum (SR) $Ca^{2+}$ signal regulates mitochondria $Ca^{2+}$ during skeletal muscle contraction by using glycolysis inhibitor. METHODS: To examine the effect of Glycolysis inhibitor on SR and mitochondria $Ca^{2+}$ content, we used skeletal muscle fiber from gastrocnemius muscle. 2-deoxy glucose and 3-bromo pyruvate used as glycolysis inhibitor, it applied to electrically stimulated muscle contraction experiment. Intracellular $Ca^{2+}$ content, SR, mitochondria $Ca^{2+}$ level and mitochondria membrane potential (MMP) was detected by confocal microscope. Mitochondrial energy metabolism related enzyme, citric acid synthase activity also examined for mitochondrial function during the muscle contraction. RESULTS: Treatment of 2-DG and 3BP decreased the muscle contraction induced SR $Ca^{2+}$ increase however the mitochondria $Ca^{2+}$ level was increased by treatment of inhibitors and showed and overloading as compared with the control group. Glycolysis inhibitor and thapsigargin treatment showed a significant decrease in MPP of skeletal muscle cells compared to the control group. CS activity significantly decreased after pretreatment of glycolysis inhibitor during skeletal muscle contraction. These results suggest that regulation of mitochondrial $Ca^{2+}$ levels by glycolysis is an important factor in mitochondrial energy production during skeletal muscle contraction CONCLUSIONS: These results suggest that mitochondria $Ca^{2+}$ level can be regulated by SR $Ca^{2+}$ level and glycolytic regulation of intraocular $Ca^{2+}$ signal play pivotal role in regulation of mitochondria energy metabolism during the muscle contraction.

• The Journal of Korean Physical Therapy
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• 제6권1호
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• pp.61-74
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• 1994

전기자극이 흰쥐의 정상근에 어떠한 영향을 미치는지를 알아보기 위하여 좌골신경의 표적근인 가자미근을 이용하여 조직계측학적, 조직화학, 전자현미경적 관찰을 하였다. 정상적으로 활동하는 흰쥐의 골격근을 매일 전기자극한 결과 전기자극군은 자극을 시작한 후 2주까지는 근섬유의 굵기와 무게가 증가하고 그 이후로는 큰 변화가 없었다. 특히 근섬유의 종류별로는 적색섬유(red muscle fiber)의 채적밀도가 증가하였고 백색섬유(white muscle fiber)는 줄어드는 양상을 보였다. 조직화학적 검사 결과 전기자극군은 근섬유가 다소비대(hypertrophy)해지고 내근주막과 외근주막 사이가 좁아져 있었다. 정상근에서는 당원이 근섬유의 뚜렷한 구별없이 양성반응을 보였으나 실험군은 특정 근섬유에만 반응이 나타났었다. 또한 NADH-TR반응 결과 전기자극군은 적색섬유가 유의하게 증가하여 생체계측학적 결과와 일치하였고 미세구조적 관찰 결과 정상근의 적색섬유는 근섬유와 평행으로 mitochondria가 형성되어 있었고 백색섬유에서는 근절부위에서 관찰되었다. 그러나 전기자극군에서는 적색섬유는 근초부위에 mitochondria가 많이 관찰되고 백색섬유도 근절부위에 작은 mitochondria의 수가 증가되는 것을 관찰할 수 있었다.

• Applied Microscopy
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• 제38권2호
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• pp.125-133
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• 2008

Mitochondria 내막의 cytochrome-c-oxidase는 세포의 에너지 생합성에 중요한 요소이며, 세포자멸사와 각종세포의 병리학적 현상과 밀접한 연관성이 있는 전자전달계효소로 알려져 있다. Porin 단백은 mitochondria 내막과 외막에 분포하는 효소단백으로 전자전달계효소 형성과 ATP 운반에 관여하는 것으로 알려져 있다. 따라서 면역현미경법을 사용하여 cytochrome-c-oxidase의 분포와 porin 단백과의 연관성을 확인하여 mitochondria의 cristae에 분포하는 cytochrome-c-oxidase의 형성과 변화를 알아보고자 하였다. Cardiac muscle tissue의 sarcoplasm에는 많은 수의 mitochondria가 분포하며, cytochrome-c-oxidase가 풍부한 mitochondria와 porin 단백이 풍부한 mitochondria로 구별되었다. Cytochrome-c-oxidase가 풍부한 mitochondria는 porin 단백이 빈약하고 porin 단백이 풍부한 mitochondria는 cytochrome-c-oxidase가 소량 포함되어 있는 것으로 관찰되었다. 심근조직의 부위에 따라 근형질에 분포하는 mitochondria에 cytochrome-c-oxidase가 풍부한 mitochondria와 porin 단백이 풍부한 mitochondria가 각각 상이하게 분포하였다. 이상의 결과로 미성숙 mitochondria는 많은 양의 porin 단백을 함유하여 근형질로부터 단백질 소단위를 mitochondria 막내로 운반하여 cytochrome-c-oxidase를 형성시키고 mitochondria가 성숙하면서 ATP를 운반할 최소한 양의 porin 단백만을 남기고 소멸되는 것으로 추측된다.

• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.567-577
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• 2017

Obesity is known to induce inhibition of glucose uptake, reduction of lipid metabolism, and progressive loss of skeletal muscle function, which are all associated with mitochondrial dysfunction in skeletal muscle. Mitochondria are dynamic organelles that regulate cellular metabolism and bioenergetics, including ATP production via oxidative phosphorylation. Due to these critical roles of mitochondria, mitochondrial dysfunction results in various diseases such as obesity and type 2 diabetes. Obesity is associated with impairment of mitochondrial function (e.g., decrease in $O_2$ respiration and increase in oxidative stress) in skeletal muscle. The balance between mitochondrial fusion and fission is critical to maintain mitochondrial homeostasis in skeletal muscle. Obesity impairs mitochondrial dynamics, leading to an unbalance between fusion and fission by favorably shifting fission or reducing fusion proteins. Mitophagy is the catabolic process of damaged or unnecessary mitochondria. Obesity reduces mitochondrial biogenesis in skeletal muscle and increases accumulation of dysfunctional cellular organelles, suggesting that mitophagy does not work properly in obesity. Mitochondrial dysfunction and oxidative stress are reported to trigger apoptosis, and mitochondrial apoptosis is induced by obesity in skeletal muscle. It is well known that exercise is the most effective intervention to protect against obesity. Although the cellular and molecular mechanisms by which exercise protects against obesity-induced mitochondrial dysfunction in skeletal muscle are not clearly elucidated, exercise training attenuates mitochondrial dysfunction, allows mitochondria to maintain the balance between mitochondrial dynamics and mitophagy, and reduces apoptotic signaling in obese skeletal muscle.

• 한국발생생물학회지:발생과생식
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• 제21권4호
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• pp.467-473
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• 2017

Ascidian embryos have become an important model for embryological studies, offering a simple example for mechanisms of cytoplasmic components segregation. It is a well-known example that the asymmetric segregation of mitochondria into muscle lineage cells occurs during ascidian embryogenesis. However, it is still unclear which signaling pathway is involved in this process. To obtain molecular markers for studying mechanisms involved in the asymmetric distribution of mitochondria, we have produced monoclonal antibodies, Mito-1, Mito-2 and Mito-3, that specifically recognize mitochondria-rich cytoplasm in cells of the ascidian Halocynthia roretzi embryos. These antibodies stained cytoplasm like reticular structure in epidermis cells, except for nuclei, at the early tailbud stage. Similar pattern was observed in vital staining of mitochondria with DiOC2, a fluorescent probe of mitochondria. Immunostaining with these antibodies showed that mitochondria are evenly distributed in the animal hemisphere blastomeres at cleavage stages, whereas not in the vegetal hemisphere blastomeres. Mitochondria were transferred to the presumptive muscle and nerve cord lineage cells of the marginal zone in the vegetal hemisphere more than to the presumptive mesenchyme, notochord and endoderm lineage of the central zone. Therefore, it is suggested that these antibodies will be useful markers for studying mechanisms involved in the polarized distribution of mitochondria during ascidian embryogenesis.

• Applied Microscopy
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• 제24권4호
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• pp.41-51
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• 1994

The topology of the enzyme has been investigated by biochemical studies including chemical labeling and cross linking. Thirteen subunits(polypeptides) of the cytochrome-c-oxidase have localistic characteristics of existing in the matrix side or cytoplasmic side in the mitochondria. In order to observe the distribution of the enzyme subunit on the mitochondria membrane, immunogold-labeling methods were employed. Antibody was obtained from the serum of immunized rabbit with enzyme subunit antigen which was obtained from cytochrome-c-oxidase of the beef heart muscle mitochondria. Beef heart muscle tissue as a tissue antigen was stained with immunized rabbit IgG and protein A gold complex. Electron microscopy has identified the existance of cytochrome-c-oxidase subunit $Mt_I,\;Mt_{II}\;and\;Mt_{III}$ on the membrane of cristae and outer chamber of mitochondria and the subunit $C_{IV}$ on the membrane of cristae and matrix of mitochondria. Particularly, the subunit $C_{IV}$ was also observed to exist in the sarcoplasm of muscle tissue.

• 대한임상전기생리학회지
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• 제1권1호
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• pp.57-72
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• 2003

This study conducts electrical stimulation to male white rat of Spargue-Dawley which is 7 weeks, has the weight of 240 g and is seemingly healthy for one or two weeks by means of neuromuscular electrical stimulator in order to examine the effects of neuromuscular electrical stimulation on its gastrocnemius, measures change of weight of gastrocnemius, serum and enzyme activity and then obtains the following conclusions. There is little difference in AST and CPK of weight and serum of gastrocnemius after one or two weeks of conducting neuromuscular electrical stimulation in all experimental groups. On the one hand, as a result of histochemical observation, NMES I group showed hypertrophy of perimysium and increase of sectional diameter of muscle fiber compared to comparison group, but NMES II group showed a similar result to comparison group. When ultrasubstructure was observed under electron microscope, I-type muscle fiber of NMES I group showed well-arranged mitochondria and it was similar to comparison group. II-type muscle fiber showed a large quantity of glycogen granules within sarcoplasmatic and the extension of luminal of T-tubule. I-type muscle fiber of NMES II group had small mitochondria and showed the vacuolar degeneration of mitochondria and extended T-tubule. II-type muscle fiber showed the extension of agranule cytoplasma reticulum with T-tubule and the reduction of amount of glycogen granule within partial sarcoplasmatic.

• Molecules and Cells
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• 제23권3호
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• pp.363-369
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• 2007

Mitochondria play a central role in energy-generating processes and may be involved in the regulation of channels and receptors. Here we investigated TRPM7, an ion channel and functional kinase, and its regulation by mitochondria. Proton ionophores such as CCCP elicited a rapid decrease in outward TRPM7 whole-cell currents but a slight increase in inward currents with pipette solutions containing no MgATP. With pipette solutions containing 3 mM MgATP, however, CCCP increased both outward and inward TRPM7 currents. This effect was reproducible and fully reversible, and repeated application of CCCP yielded similar decreases in current amplitude. Oligomycin, an inhibitor of $F_1/F_O$-ATP synthase, inhibited outward whole-cell currents but did not affect inward currents. The respiratory chain complex I inhibitor, rotenone, and complex III inhibitor, antimycin A, were without effect as were kaempferol, an activator of the mitochondrial $Ca^{2+}$ uniporter, and ruthenium red, an inhibitor of the mitochondrial $Ca^{2+}$ uniporter. These results suggest that the inner membrane potential (as regulated by proton ionophores) and the $F_1/F_O$-ATP synthase of mitochondria are important in regulating TRPM7 channels.

• Journal of Nutrition and Health
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• 제54권4호
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• pp.335-341
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• 2021

Purpose: Muscle mitochondria play a key role in regulating fatty acid and glucose metabolism. Dysfunction of muscle mitochondria is associated with metabolic diseases such as obesity and type 2 diabetes. Isorhamnetin (ISOR), also known as 3-O-methylquercetin, a quercetin metabolite, is a naturally occurring flavonoid in many plants. This study evaluated the effects of ISOR on the regulation of the mitochondrial function of C2C12 muscle cells. Methods: C2C12 muscle cells were differentiated for 5 days, and then treated in various concentrations of ISOR. Cytotoxicity was determined by assessing cell viability using the water-soluble tetrazolium salt-8 assay principle at different concentrations of ISOR and time points. Levels of the mitochondrial DNA (mtDNA) content and gene expression were measured by quantitative real-time polymerase chain reaction. The citrate synthase (CS) activity was quantified by the enzymatic method. Results: ISOR at a concentration of 10 µM did not show any cytotoxic effects. ISOR increased the mtDNA copy number in a time- or dose-dependent manner. The messenger RNA levels of genes involved in mitochondrial function, such as peroxisome proliferator-activated receptor-γ coactivator-1α, and uncoupling protein 3 were significantly stimulated by the ISOR treatment. The CS activity was also significantly increased in a time- or dose-dependent manner. Conclusion: These results suggest that ISOR enhances the regulation of mitochondrial function, which was at least partially mediated via the stimulation of the mtDNA replication, mitochondrial gene expression, and CS activity in C2C12 muscle cells. Therefore, ISOR may be useful as a potential food ingredient to prevent metabolic diseases-associated muscle mitochondrial dysfunction.

• The Journal of Korean Physical Therapy
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• 제14권2호
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• pp.145-152
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• 2002

고압맥동전류가 탈신경근의 형태에 미치는 영향을 알아보기 위하여 조직화학적 방법과 투과전자현미경적 관찰을 하였다. 웅성 흰쥐를 정상군, 탈신경군으로 각각 8마리씩 나누어 2주와 4주 후에 희생시켜 실험한 결과 다음과 같은 결과를 얻었다. 1. 조직화학적으로 형태를 관찰 한 결과 대조 1주군부터 근속과 근섬유사이에 염증세포가 관찰되고 핵이 근섬유속에 위치하는 것도 자주 관찰되었다. 2. 대조군도 비슷한 양상을 보였으나 4주군은 근괴사와 염증세포가 더욱 증가하였다. 3. 당원 1주군에서 실험군, 대조군 모두 정상근과 비슷한 양상을 보이나 4주군에서는 섬유를 구별할 수 없는 형태로 관찰되었다. 4. NADH-TR반응에서 적색섬유가 2주군에서 약간 증가하였고 그 후로는 구별이 불가능하였다. 5. 미세구조적으로 양쪽군 모두 근섬유가 굽어있고 mitochondria의 파괴로 인한 공포가 많이 관찰되었으나 전기자극 2주군에서는 일부에서 mitochondria증가를 관찰 하였다.