• 대한의생명과학회지
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• 제10권4호
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• pp.385-389
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• 2004

A novel gene Jpk, originally isolated as a trans-acting factor associating with the position-specific regulatory element of murine Hox gene has been reported to be expressed differentially in the liver of diabetic animals. Therefore, in an attempt to develop a possible diagnostic marker and/or new therapeutic agent for the Diabetes Mellitus, we analysed the expression pattern of Jpk among organs of normal and diabetic Sprague-Dawley (SD) rats. Total RNAs were isolated from each organs (brain, lung, heart, liver, spleen, kidney, muscle, blood, and testis) of diabetic and normal rats in both normal feeding and after fasting condition. And then RT (reverse transcription) PCR has been performed using Jpk­specific primers. The Jpk gene turned out to be expressed in all organs tested, with some different expression profiles among normal and diabetes, though. Upon fasting, Jpk expressions were reduced in all organs tested except kidney, muscle and brain of normal rat. Whereas in diabetes, Jpk expressions were increased in all organs except heart, muscle and testis when fasted. Compared to the normal rat, the Jpk expression level in blood was remarkably upregulated (about 15-30times) in diabetic rat whether in normal feeding or fasting conditon, suggesting that the Jpk could be a candidate gene for the possible blood diagnostic marker for the Diabetes Mellitus.

• International Journal of Stem Cells
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• 제16권2호
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• pp.145-155
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• 2023

Background and Objectives: Embryologically, mesodermal development is closely related to the development of various organs such as muscles, blood vessels, and hearts, which are the main organs that make up the body. However, treatment for mesoderm developmental disorders caused by congenital or acquired factors has so far relied on surgery and drug treatment for symptom relief, and more fundamentally, treatment for mesoderm developmental disorders is needed. Methods and Results: In our study, microRNA (miRNA), which plays an important role in the mesoderm development process, was identified and the developmental function was evaluated. miRNAs consist of small nucleotides, which act as transcription factors that bind to the 3' untranslated region and suppressed target gene expression. We constructed the human embryonic stem cell (hESC) knockout cell line and analyzed the function and characteristics of miR-5739, which plays an important role in mesoderm lineage. miR-5739 acts as a transcription factor targeting SMA, Brachyury T, Hand1, which controls muscle proliferation and differentiation, and KDR gene, which regulates vessel formation in vitro. In vivo results suggest a role in regulating muscle proliferation and differentiation. Gene ontology analysis confirmed that the miR-5739 is closely related to genes that regulate muscle and vessel proliferation and differentiation. Importantly, abnormal expression of miR-5739 was detected in somatic cells derived from patients with congenital muscle disease. Conclusions: Our study demonstrate that miR-5739 gene function significantly affects transcriptional circuits that regulate muscle and vascular differentiation during embryonic development.

• 한국어류학회지
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• 제23권1호
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• pp.1-9
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• 2011

ACP (annealing control primer)를 사용하여 DDRT (differential display reverse transcription)-PCR 방법으로 볼락의 성장단계에 따라 발현량 차이를 나타내는 DEG (differentially expressed gene)를 확보하였다. ACP 120개를 분석하여 18개월령 근육조직에서보다 6개월령 근육조직에서 발현량이 많은 DEG 16개와 6개월령 근육조직에서보다 18개월령 근육조직에서 발현량이 더 많은 DEG22개의 염기서열을 분석하였다. DEG 염기서열을 BLAST 검색한 결과, parvalbumin (PVALB) 등 18개의 유전자(PVALB, NDKB, TPM, TnI, GAPDH, CKM2, factor 2 SERF2, AMPD, TRICA, ARHGAP15, ESD, hsp70, COL1A2, GST, Midllip1, MYL1, SERCA1B, FTH1)와 69~95%의 상동성을 나타냈다. Real time PCR 분석법으로 6개월령 근육조직에서 발현량이 많은 DEG14와 PVALB 유전자의 성장단계별 발현양상을 조사한 결과, 볼락이 성장함에 따라 발현량이 감소하였으며, 특히 PVALB 유전자는 6개월령 이후에는 발현량이 극히 적었다. 6개월령 근육조직에서보다 18 개월령 근육조직에서 발현량에서 많았던 CKM2 유전자는 성장함에 따라 발현량이 계속 증가하였고, 4세 이후에는 발현량이 감소하였다. DEG의 조직특이적 발현양상을 분석한 결과, DEG14는 근육, 간, 신장, 및 비장조직에서 발현되었으며, PVALB 유전자는 근육과 신장조직에서 발현되었고, 간과 비장조직에서는 발현되지 않았다. CKM2 유전자는 근육, 신장 및 비장조직에서 발현되었고, 간 조직에서는 발현되지 않았다. PVALB 유전자의 mRNA 크기는 659 bp 이며, 110개의 아미노산으로 구성되어 있다. Parvalbumin과 CKM2 유전자는 성장속도가 빠른 어류 선발에 이용할 수 있는 분자마커 개발에 활용하고자한다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권10호
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• pp.1365-1373
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• 2013

Fat mass and obesity associated gene (FTO) plays an important role in appetite control and energy consumption in human and mice. In order to examine FTO expression influence on fat deposition in Suzhong pigs, FTO mRNA expression was detected in 16 tissues by RT-PCR, FTO protein expression was detected in 5 tissues by western blot, and association of FTO polymorphism with meat quality traits was analyzed in Suzhong populations with 714 records. RT-PCR results revealed that FTO mRNA was expressed in all sixteen tissues with significant differences (p<0.05), expression in backfat was significantly higher than that of any other tissue (p<0.05), and expression in longissimus dorsi muscle had the second highest significance level (p<0.05). Western blot results demonstrated that FTO protein was highly expressed in backfat and longissimus dorsi muscle. Furthermore, FTO mRNA and protein expression in tissues of high-fat pigs was significantly higher than that of low-fat pigs (p<0.05), suggesting FTO expression had advantageous effects on fat deposition. FTO polymorphism results evidenced that at A227G locus, G allele seemed to have advantageous effects on fat deposition, indicating it could be a significant candidate gene for improving pork quality in Suzhong pigs.

• The Korean Journal of Physiology and Pharmacology
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• 제21권1호
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• pp.37-44
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• 2017

Regulation of vascular smooth muscle cell (VSMC) phenotype plays an essential role in many cardiovascular diseases. In the present study, we provide evidence that $kr{\ddot{u}}ppel$-like factor 8 (KLF8) is essential for tumor necrosis factor ${\alpha}$ ($TNF{\alpha}$)-induced phenotypic conversion of VSMC obtained from thoracic aorta from 4-week-old SD rats. Stimulation of the contractile phenotype of VSMCs with $TNF{\alpha}$ significantly reduced the VSMC marker gene expression and KLF8. The gene expression of KLF8 was blocked by $TNF{\alpha}$ stimulation in an ERK-dependent manner. The promoter region of KLF8 contained putative Sp1, KLF4, and $NF{\kappa}B$ binding sites. Myocardin significantly enhanced the promoter activity of KLF4 and KLF8. The ectopic expression of KLF4 strongly enhanced the promoter activity of KLF8. Moreover, silencing of Akt1 significantly attenuated the promoter activity of KLF8; conversely, the overexpression of Akt1 significantly enhanced the promoter activity of KLF8. The promoter activity of SMA, $SM22{\alpha}$, and KLF8 was significantly elevated in the contractile phenotype of VSMCs. The ectopic expression of KLF8 markedly enhanced the expression of SMA and $SM22{\alpha}$ concomitant with morphological changes. The overexpression of KLF8 stimulated the promoter activity of SMA. Stimulation of VSMCs with $TNF{\alpha}$ enhanced the expression of KLF5, and the promoter activity of KLF5 was markedly suppressed by KLF8 ectopic expression. Finally, the overexpression of KLF5 suppressed the promoter activity of SMA and $SM22{\alpha}$, thereby reduced the contractility in response to the stimulation of angiotensin II. These results suggest that cross-regulation of KLF family of transcription factors plays an essential role in the VSMC phenotype.

• The Korean Journal of Physiology and Pharmacology
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• 제22권4호
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• pp.427-436
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• 2018

The objective of this study was to analyze the concurrent treatment effects of ursolic acid (UA) and low-intensity treadmill exercise and to confirm the effectiveness of UA as an exercise mimetic to safely improve muscle atrophy-related diseases using Sprague-Dawley (SD) rats with skeletal muscle atrophy. Significant muscle atrophy was induced in male SD rats through hind limb immobilization using casting for 10 days. The muscle atrophy-induced SD rats were group into four: SED, sedentary; UA, daily intraperitoneal UA injection, 5 mg/kg; EX, low-intensity (10-12 m/min, $0^{\circ}$ grade) treadmill exercise; and UEX, daily intraperitoneal UA injection, 5 mg/kg, and low-intensity (10-12 m/min, $0^{\circ}$ grade) treadmill exercise. After 8 weeks of treatment, endurance capacity was analyzed using a treadmill, and tissues were extracted for analysis of visceral fat mass, body weight, muscle mass, expression of muscle atrophy- and hypertrophy-related genes, and endurance capacity. Although the effects of body weight gain control, muscle mass increase, and endurance capacity improvement were inadequate in the UA group, significant results were confirmed in the UEX group. The UEX group had significantly reduced body weight and visceral fat, significantly improved mass of tibialis anterior and gastrocnemius muscles, and significantly decreased atrophy-related gene expression of MuRF1 and atrogin-1, but did not have significant change in hypertrophy-related gene expression of Akt and mTOR. The endurance capacity was significantly improved in the EX and UEX groups. These data suggest that concurrent treatment with low-intensity exercise and UA is effective for atrophy-related physical dysfunctions.

• The Journal of Korean Physical Therapy
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• 제15권4호
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• pp.190-198
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• 2003

Expression of c-fos, an immediate early gene, has accepted to be a marker of functional activity in neurons. This study was aimed to investigate the effects of cold therapy on the expression of spinal cord c-fos in rats of carrageenan-induced muscle pain. Muscle pain was induced in male Sprague-Dawley rats by intra-muscular injection of gastrocnemius with $2\%$ carrageenan. The paw withdrawal latency (PWL) and tail flick test (TFT) responses to heat stimuli were used to detect secondary hyperalgesia produced by the muscle pain and measured to assess the effects of cold. The expression of c-fos was determined in the lumbar regions of the spinal cord by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry assays. The secondary hyperalgesia to heat simuli (PWL and TFT) were significantly reduced in cold therapy compared with that in the controls. In RT-PCR assays the expression of c-fos mRNA was down-regulated in the lumbar spinal cord in cold group. In addition, Fos immunoreactivity in the dorsal horn of the lumbar spinal cord was decreased in cold group. These results suggested that application of cold attributed to increase PWL and TFT responses and to decrease expression of the c-fos produced by muscle pain.

• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.205-221
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• 2004

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

• Asian-Australasian Journal of Animal Sciences
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• 제28권5호
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• pp.697-702
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• 2015

While athletic abilities such as speed, endurance and recovery are important in the horse, genes related to these abilities have not been extensively investigated. Here, we characterized the horse peroxisome proliferator-activated receptor delta ($PPAR{\delta}$) gene and analyzed the expression of $PPAR{\delta}$ during exercise. $PPAR{\delta}$ is a known regulator of ${\beta}$-oxidation, muscle fiber transformation, and running endurance. Through evolutionary analysis using the synonymous and non-synonymous mutation ratio, it was revealed that positive selection occurred in the horse $PPAR{\delta}$ gene. Two important domains related to nuclear hormone receptors, C4 zinc finger and ligand binding domain, were also found to be conserved well in horse $PPAR{\delta}$. Horse $PPAR{\delta}$ was expressed ubiquitously in many tissues, but the expression level was various depending on the tissues. In the skeletal muscle, $PPAR{\delta}$ increased about 2.5 folds after 30 min of exercise. Unlike in muscle, the increase of $PPAR{\delta}$ expression was observed at 60 min but not 30 min of exercise in leukocytes. This finding might be useful for testing the endurance of horse using blood samples. Conclusively, the horse $PPAR{\delta}$ gene is evolutionarily conserved well and can be used as a biomarker of endurance in horse.

• 약학회지
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• 제45권1호
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• pp.108-115
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• 2001

We have recently reported the development of a high efficiency expression vector, pCK, which can drive a high level of gene expression in mouse skeletal muscle. In this study, we tested the therapeutic potential of pCK expressing human VEGF165, pCK-VEGF in the rabbit ischemic hind limb model. To determine the optimal dose of plasmid DNA, various concentrations of pCK-CAT were injected into the muscle of a rabbit hind limb and the levels of CAT activity were determined. It was found that the expression level of the exogenously added gene became stable between 250 and 1,000 $\mu$g. Based on this result, we tested whether intramuscular transfer of 500$\mu$g of pCK-VEGF could actually modulate collateral vessel development in a rabbit ischemic hind limb model. It was found that relative to the control group injected with the pCK lacking the VEGF sequence, single intramuscular doses (500$\mu$g) of pCK-VEGF produced statistically significant augmentation of collateral vessels as determined by the angiographic vessel count, maximal blood flow by Doppler flowmeter and the number of capillaries by histology. These results suggest that a single 500$\mu$g-delivery of pCK-VEGF is potent enough to induce sufficient angiogenic activity and achieve therapeutic benefit on this rabbit model.