• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1288-1302
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• 2011

The aim of this study was to analyze the proteome expression of bovine satellite cells from longissimus dorsi (LD), deep pectoral (DP) and semitendinosus (ST) muscle depots during in vitro myogenic differentiation. Proteomic profiling by twodimensional gel electrophoresis and mass spectrometry of differentiating satellite cells revealed a total of 38 proteins that were differentially regulated among the three depots. Among differentially regulated proteins, metabolic proteins like lactate dehydrogenase (LDH), malate dehydrogenase (MDH) were found to be up regulated in ST, while alpha-enolase (NNE) in LD and DP depot satellite cells were down regulated. Also, our analysis found that there was a prominent up regulation of cytoskeletal proteins like actin, actincapping protein and transgelin along with chaperone proteins like heat shock protein beta 1 (HSPB 1) and T-complex protein 1 (TCP-1). Among other up regulated proteins, LIM domain containing protein, annexin 2 and Rho GDP-dissociation inhibitor 1 (Rho GDI) are observed, which were already proven to be involved in the myogeneis. More interestingly, satellite cells from ST depot were found to have a higher myotube formation rate than the cells from the other two depots. Taken together, our results demonstrated that, proteins involved in glucose metabolism, cytoskeletal modeling and protein folding plays a key role in the myogenic differentiation of bovine satellite cells.

• Genomics & Informatics
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• 제2권2호
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• pp.67-74
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• 2004

Cells consistently face stressful conditions, which cause them to modulate a variety of intracellular processes and adapt to these environmental changes via regulation of gene expression. Hyperosmotic and oxidative stresses are significant stressors that induce cellular damage, and finally cell death. In this study, oligonucleotide microarrays were employed to investigate mRNA level changes in cells exposed to hyperosmotic or oxidative conditions. In addition, since heat shock protein 70 (HSP70) is one of the most inducible stress proteins and plays pivotal role to protect cells against stressful condition, we performed microarray analysis in HSP70-overexpressing cells to identify the genes expressed in a HSP70-dependent manner. Under hyperosmotic or oxidative stress conditions, a variety of genes showed altered expression. Down­regulation of protein phosphatase1 beta (PP1 beta) and sphingosine-1-phosphate phosphatase 1 (SPPase1) was detected in both stress conditions. Microarray analysis of HSP70-overexpressing cells demonstrated that diverse mRNA species depend on the level of cellular HSP70. Genes encoding Iysyl oxidase, thrombospondin 1, and procollagen displayed altered expression in all tested conditions. The results of this study will be useful to construct networks of stress response genes.

• 원예과학기술지
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• 제31권3호
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• pp.275-281
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• 2013

Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.28-41
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• 2024

Objective: Chronic scrotal hyperthermia (SHT) can lead to serious disorders of the male reproductive system, with oxidative stress playing a key role in the onset of these dysfunctions. Thus, we evaluated the impact of caffeine, a potent antioxidant, on cellular and tissue disorders in mice with chronic SHT. Methods: In this experimental study, 56 adult male NMRI mice were allocated into seven equal groups. Apart from the non-treated control group, all were exposed to heat stress. Two groups, termed "preventive" and "curative," were orally administered caffeine. The preventive mice began receiving caffeine immediately prior to heat exposure, while for the curative group, a caffeine regimen was initiated 15 consecutive days following cessation of heat exposure. Each treated group was subdivided based on pairing with a positive control (Pre/ curative [Cur]+PC) or a vehicle (Pre/Cur+vehicle). Upon conclusion of the study, we assessed sperm characteristics, testosterone levels, stereological parameters, apoptosis, antioxidant and oxidant levels, and molecular markers. Results: Sperm parameters, testosterone levels, stereological parameters, biochemical factors (excluding malondialdehyde [MDA]), and c-kit gene expression were significantly elevated in the preventive and curative groups, especially the former, relative to the other groups. Conversely, expression levels of the heat shock protein 72 (HSP72) and nuclear factor kappa beta (NF-κβ) genes, MDA levels, and apoptotic cell density were markedly lower in both caffeine-treated groups relative to the other groups, with more pronounced differences observed in the preventive group. Conclusion: Overall, caffeine attenuated cellular and molecular abnormalities induced by heat stress in the testis, particularly in the mice treated under the preventive condition.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.946-951
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• 2006

To study the effect of iron on Saccharomyces cerevisiae, whole-cell proteins of Saccharomyces cerevisiae were extracted and subjected to two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and differentially expressed proteins were identified. The proteins separated were further identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and were compared with a protein database. Of more than 300 spots separated by molecular weight and isoelectric points, 27 differentially expressed spots were identified. Ten proteins were found to be differentially expressed at high iron concentration. Triosephosphate isomerase (TPI), YDR533C hypothetical protein, superoxide dismutase (SOD), 60 kDa heat-shock protein (HSP60), pyruvate dehydrogenase beta subunit 1 (PDB1), and old yellow enzyme 2 (OYE2) were upregulated, whereas thiol-specific antioxidant (TSA), regulatory particle non-ATPase subunit 8 (RPN8), thiol-specific peroxiredoxin 1 (AHP1), and fructose-1, 6-bisphosphate adolase (FBA) were downregulated by iron. Based on the result, we propose that SOD upregulated by iron would protect the yeast from oxidative stress by iron, and that TSA downregulated by iron would render cells hypersensitive to oxidative stress.

• 생명과학회지
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• 제23권4호
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• pp.586-594
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• 2013

단백질 번역 후 O-GlcNAc 수식은 단백질 조절의 새로운 기전으로 대두되고 있다. 전통적인 당수식과 달리 O-GlcNAc 수식은 단 한번의 O-GlcNAc 전달로 이루어지며, 핵 및 세포질단백질 모두에 수식될 수 있다. O-GlcNAc은 이 분자를 끝으로 하는 최종수식으로 생각되어 왔으나, 최근의 논문(J Proteome Res. 2011 10:2725-2733)은 AP180 단백질에 O-GlcNAc-P가 존재함을 보고하였다. 이 논문은 O-GlcNAc-P가 일반적인 단백질수식인지에 대한 중요한 질문을 던진다. 이에 답하고자 저자들은 HEK293T 세포에 O-GlcNAc 인산화효소 NAGK를 DsRed2에 연결한 DsRed2-$NAGK_{WT}$ 혹은 효소활성이 없는 돌연변이 NAGK를 표현하는 DsRed2-$NAGK_{D107A}$를 표현시키고, 단백질 추출물을 얻어 2D-PAGE로 분리한 후 인산화 정도를 측정하여, $NAGK_{WT}$에 의하여 인산화가 증가되는 15개의 단백질 스폿을 선별하였다. 이 가운데 7개 스팟을 동정한 결과 2개의 스폿은 O-GlcNAc 수식 단백질인 $HSP90{\beta}$, 다른 2개의 스폿도 O-GlcNAc 수식 단백질인 ENO1로 동정되었으며, 나머지(dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P, grp94)는 O-GlcNAc 수식 여부를 아직 모르는 단백질이였다. NAGK에 의하여 O-GlcNAc 단백질의 인산화가 증가된다는 사실은 O-GlcNAc이 인산화되어 O-GlcNAc-P로 수식됨을 시사하며, 따라서 본 연구의 결과는 O-GlcNAc이 최종 수식이 아님을 지지한다.

• Molecular & Cellular Toxicology
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• 제4권3호
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• pp.199-204
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• 2008

The bio-molecules involved in defense mechanisms can be used as efficient biomarkers for physiological changes in organisms caused by both of internal and external stress. Thus, the expression level of genes which encoding such molecules serve as critical 'early warning system' for environmental assessment as well as health diagnosis of biological organisms. In this study, Cytochrome P450, Heat shock protein 90, Ubiquitin and ${\beta}$-actin gene were isolated for the first time from a red alga Gracilaria textorii. The quantitative differential gene expression analyses of three genes, GteCYP1A, GteHsp90 and Gte-UB, were carried out in G. textorii sporophytes collected from two different localities, polluted Sujeong (Masan, Korea) and potentially unpolluted Danggeum (Daemaemuldo Is., Korea). The transcripts of all three tested genes were highly expressed in the Sujeong population. The results suggest: 1) the Sujeong site was more polluted than the Danggeum site; 2) G. textorii could be applicable to marine environment monitoring in coastal regions.

• Reproductive and Developmental Biology
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• 제33권4호
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• pp.243-248
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• 2009

To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

• Journal of Acupuncture Research
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• 제22권5호
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• pp.57-66
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• 2005

Objectives : We examined the effects of electroacupuncture on the cholecystokinin-octapeptide-induced acute pancreatitis in rats. Methods : Rats were administered with $75{\mu}g/kg$ cholecystokinin-octapeptide subcutaneously three times (1, 3 and 5h after shaving) for 5days. Three days after finishing cholecystokinin-octapeptide administration, high frequency electroacupuncture (100Hz) and low frequency electroacupuncture (2Hz) were applied to acupoint equivalent to ST36 (Zusanli) for 7 days. The author determined the pancreatic weight/body weight ratio, the levels of pancreatic heat shock protein HSP60 and HSP72. The author also assay the secretion of ${\beta}-amylase$, lipase and pro-inflammatory cytokines in serum. Repeated cholecysokinin-octapeptide treatment resulted in the typical laboratory and morphological changes of experimentally induced pancreatitis. Results : Eelectroacupuncture significantly decreased the pancreatic weight/body weight ratio in cholecystokinin-octapeptide-induced acute pancreatitis, increased the pancreatic levels of HSP60 and HSP72, and decreased ${\beta}-amylase$ and lipase levels in cholecystokinin-octapeptide-induced acute pancreatitis. Additionally, the secretion of $Interleukin-1{\beta}$ and tumor necrosis $factor-{\alpha}$ was decreased in the animals treated with electroacupuncture. Conclusion : These results suggest that electroacupuncture may have protective effects against cholecystokinin-octapeptide-induced acute pancreatitis.

• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.749-756
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• 2014

The present study was carried out to investigate the effects of dietary antioxidants on pro-inflammatory cytokines, heat shock protein (HSP) and antioxidant status in broiler chicks under summer conditions. A total of 162, 3-d-old broiler chicks were randomly assigned to a basal diet (CON) and the basal diet supplemented with vitamin C (200 mg/kg diet, VCD) or vitamin E (100 mg/kg, VED) until 35 day of age. All birds were exposed to summer diurnal heat stress at average daily fluctuations of temperature between $32^{\circ}C$ to $34^{\circ}C$ at day to $27^{\circ}C$ to $29^{\circ}C$ at night for the entire feeding periods. There was no significant difference in body weight, feed to gain ratio and the relative organ weight except the thymus in response to dietary vitamin C or E supplementation. However, the mRNA expression of interleukin (IL)-$1{\beta}$, IL-6, interferon (IFN)-${\gamma}$, Toll like receptor (TLR)-4 and HSP70 in the liver of birds fed diet containing vitamin C significantly (p<0.05) decreased compared with those in birds fed basal diet. Dietary vitamin E also showed a significant (p<0.05) decrease in the mRNA expression of IL-6 and HSP70 compared with a basal diet. Total antioxidant status (TAS) in serum of birds fed vitamin C supplemented diet was significantly (p<0.05) higher with than that in birds a basal diet. Lipid peroxidation in serum and liver resulted in a significant (p<0.05) decrease in response to dietary vitamin C or E supplementation. In conclusion, dietary supplementation with antioxidant vitamins, especially vitamin C resulted in a significant decrease in the mRNA expression of pro-inflammatory cytokines and HSP70, and higher antioxidant parameters than that of birds on the basal diet under summer conditions.