• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.4
• /
• pp.457-463
• /
• 2016

The goal of this work was to investigate the correlations between MyHC mRNA transcription and their corresponding protein expressions in porcine longissimus muscle (LM) during postnatal growth of pigs. Five DLY ($Duroc{\times}Landrace{\times}Yorkshire$) crossbred pigs were selected, slaughtered and sampled at postnatal 7, 30, 60, 120, and 180 days, respectively. Each muscle was subjected to quantity MyHCs protein contents through an indirect enzyme-linked immunosorbent assay (ELISA), to quantity myosin heavy-chains (MyHCs) mRNA abundances using real-time polymerase chain reaction. We calculated the proportion (%) of each MyHC to total of four MyHC for two levels, respectively. Moreover, the activities of several key energy metabolism enzymes were determined in LM. The result showed that mRNA transcription and protein expression of MyHC I, IIa, IIx and IIb in LM all presented some obvious changes with postnatal aging of pigs, especially at the early stage after birth, and their mRNA transcriptions were easy to be influenced than their protein expressions. The relative proportion of each MyHC mRNA was significantly positively related to that of its corresponding protein (p<0.01), and MyHC I mRNA proportion was positively correlated with creatine kinase (CK), succinate dehydrogenase (SDH), malate dehydrogenase (MDH) activities (p<0.05). These data suggested that MyHC mRNA transcription can be used to reflect MyHC expression, metabolism property and adaptive plasticity of porcine skeletal muscles, and MyHC mRNA composition could be a molecular index reflecting muscle fiber type characteristics.

• Korean Journal of Food Science and Technology
• /
• v.28 no.2
• /
• pp.292-297
• /
• 1996

In this study, biochemical changes of muscle protein in squid Sikhae prepared at various temperatures and moisture content were investigated. The pH values in squid Sikhae decreased rapidly with increase in titratable acidity during fermentation at high temperature and moisture content. Growth rate of lactic acid bacteria increased rapidly during fermentation at high temperature and moisture content. Changes in muscle protein subunit were analyzed by SDS-PAGE; myosin heavy chain component disappeared completely during preparation of squid Sikhae and actin component decreased slowly during fermentation. Therefore, it was suggested that myosin heavy chain component in squid muscle protein was autolyzed at the incipient for-mentation time and actin component was degraded by acidic protease in muscle and protease produced from microorganisms during fermentation.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.29 no.3
• /
• pp.296-308
• /
• 1996

Proteolytic enzymes responsible for post-mortem degradation of the fish tissues have been studied in regard with screening the proteases distributed in the fish body by reacting with the specific synthesized substrates. Activities of cathepsin L, B, H, G, and D like enzymes were detected in the muscle crude protease from the both kind of fish, dark fleshed fish (anchovy, Engraulis japonica, and gizzard-shad, Clupanodo punctatus) and white fleshed fish (seabass, Lateolabrax japonicus, and sole, Pleuronichthys cornutus), however, those of chymotrypsin, trypsin, pepsin, and peptidase like enzymes were observed 3n the viscera crude pretense from the fish. Proteolytic activities of the muscle crude protease at pH 6.0 were similar to those of the viscera crude protease at pH 8.0, but, those of the viscera crude protease at pH 8.0 were about 2 times higher than those at pH 6.0. The muscle and viscera crude protease from anchovy showed the strongest proteolytic activity among the four fish crude proteases and the proteolytic activity of the viscera crude protease was approximately 100 times higher than that of the muscle crude protease, which suggest that viscera proteases were more contributed on the development of post-mortem changes than muscle proteases. With the degradation patterns on SDS-polyacrylamide gel electrophoresis against yellowtail myofibrillar proteins, the muscle and viscera crude protease of the four fishes were primary responsible for the degradation of myosin heavy chain, and myosin light chain and actin, respectively.

• Korean Journal of Food Science and Technology
• /
• v.30 no.4
• /
• pp.883-887
• /
• 1998

Protease activities were measured in vegetables used as cooking material or a side dish. Proteases of green pepper (Kwari and Chungyang), perilla leaves, soybean sprout and mungbean sprout were showed high activities. Especially, protease in soybean sprout was the highest activity among them. After dialysis, remaining activities in mungbean sprout, green pepper (Kwari and Chungyang) and perilla leaves were 12, 23, 45% and 37%. In the results of thawing after freezing the proteases, remaining activities in sprout of mungbean and soybean were 100% and 65%. Protease in soybean sprout was showed higher activity and stability than others. Proteolytic effects of soybean sprout on myofibrillar and sarcoplasmic protein were showed higher than stroma protein. In SDS-PAGE, myosin heavy chain, actin and tropomyosin were hydrolyzed with increasing time. Protease activity was rapidly increased with increasing meat concentration in a early proteolysis reaction, but was slightly increased in later.

• Journal of Korean Medicine Rehabilitation
• /
• v.19 no.1
• /
• pp.11-21
• /
• 2009

Objectives : This study was performed to investigate the effects of Semen Persicae(SP) on muscle type change and vascularization effect by measuring expression of Myosin heavy chain (MHC) and Vascular endothelial growth factor (VEGF) protein in the Middle cerebral artery occlusion(MCAo) rats. Methods : The middle cerebral artery was occluded, SP extraction was administrated for 4 days. The effects of SP on muscle type change and vascularization were measured. Results : 1. VEGF protein expressions in the Semen Persicae oral administration group of MCAo group were increased compared to the control group. 2. There were no significant difference between the Semen Persicae oral administration of MCAo group and the control group in MHC isoform (Type I, Type IIa) expression change. Conclusions : The present study demonstrates the effect of Semen Persicae in the vascularizing after ischemia, but has no significant effect in musle type change and the improvement of musle atrophy.

• Korean Journal of Veterinary Service
• /
• v.31 no.4
• /
• pp.449-456
• /
• 2008

The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

• Korean Journal of Food Science and Technology
• /
• v.22 no.6
• /
• pp.640-645
• /
• 1990

Changes of the electrophoretic patterns of myofibrillar proteins by sugar audition and heat treatment was studied. In the electrophoretic patterns of myofibrills prepared from no sugar added meat, as the intensity of higher molecular weight band such as myosin heavy chain showed a remarkable decrease by heating, that of lower molecular weight band such as actin showed no change. That from sugar added meat showed more remarkable decrease in the intensity of higher molecular weight band than that from no sugar added meat and this tendency was most noticeable in case of glucose addition. The effect of digestion with proteases after sugar addition and heat treatment on the electrophoretic patterns exhibited the descending order of trypsin >chymotrypsin >peptidase. By digestion with these three enzymes at one time myosin produced 27.000 dalton and 32.000 dalton components, and actin showed 16,000 dalton component. in the case of heat treatment, a part of actin was not digested. And in the case of glucose addition the myosin aggregates was not digested with these three enzymes at a time.

• Animal cells and systems
• /
• v.14 no.1
• /
• pp.25-30
• /
• 2010

We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

• Molecules and Cells
• /
• v.25 no.1
• /
• pp.78-85
• /
• 2008

In a search for new molecular pathways associated with asthma, we performed an mRNA differential display analysis using total RNA extracted from the tracheal tissues of ovalbumin (OVA)-challenged mice and sham controls. cDNAs corresponding to mRNAs for which expression levels were altered by OVA-challenge were isolate and sequenced. Twenty-eight genes differentially expressed in sham and OVA challenged mice were identified. A GenBank BLAST homology search revealed that they were related to cytoskeleton remodeling, transcription, protein synthesis and modification, energy production, and cell growth and differentiation. Two were selected for further characterization. Up-regulation of both the perinatal skeletal myosin heavy chain (skMHC) and fast skeletal muscle myosin light chain (skMLC) genes was confirmed by RT-PCR of trachea tissue from OVA challenged mice. Overexpression of skMLC protein was observed in the smooth muscle layers of OVA-challenged mice by immunohistochemistry, and the surface areas stained with skMLC antibody increased in the OVA-challenged mice. The overexpression of skMLC in murine asthma may be associated with the changes of bronchial smooth muscle.

• Food Science of Animal Resources
• /
• v.43 no.2
• /
• pp.359-373
• /
• 2023

This study examined the α-glucosidase inhibitory, and apoptosis- and anti-muscular-related factors of goat meat extracts from forelegs, hind legs, loin, and ribs. The goat meat extracts were evaluated for their α-glucosidase inhibitory activity. The gene and protein expression levels of Bcl-2-associated X (bax), p53, and p21 were examined by reverse transcription polymerase chain reaction (RT-PCR) and immunoblotting in AGS and HT-29 cells. The expression levels of Atrogin-1 and MHC1b were examined by RT-PCR in C2C12 myoblasts, and the expression levels of Atrogin-1, muscle atrophy F-box (MAFbx), muscle RING-finger protein-1 (MuRF-1), and myosin heavy chain-7 were investigated by immunoblotting. α-Glucosidase inhibitory activity was higher in ethanol extract than in hydrous and hot water extracts. BAX and p53 expression levels were higher (p<0.05) in AGS cells treated with goat meat extract than those of cells treated with no goat meat extract. In HT-29 cells, the protein expression levels of BAX, p53, and p21 were higher (p<0.05) in the cells treated with goat meat extract than those of cells not treated with goat meat extract. In dexamethasone-treated C2C12 cells, goat meat extract treatment lower (p<0.05) the expression of Atrogin-1 and lower (p<0.05) the expression of MAFbx and MuRF-1. The results of the present study indicate that goat meat extracts have α-glucosidase inhibitory activity in vitro. In addition, apoptosis was induced in AGS cells and HT-29 cells treated with goat meat extract, and anti-muscular atrophy activity was also observed in C2C12 cells treated with goat meat extract.