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Proteomic Analysis of Bovine Muscle Satellite Cells during Myogenic Differentiation
Rajesh, Ramanna Valmiki,Jang, Eun-Jeong,Choi, In-Ho,Heo, Kang-Nyeong,Yoon, Du-Hak,Kim, Tae-Hun,Lee, Hyun-Jeong Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.9
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.
Rajesh, Ramanna Valmiki,Kim, Seong-Kon,Park, Mi-Rim,Nam, Jin-Seon,Kim, Nam-Kuk,Kwon, Seulemina,Yoon, Du-Hak,Kim, Tae-Hun,Lee, Hyun-Jeong Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.1
Anatomically separate fat depots differ in size, function, and contribution to pathological states such as the metabolic syndrome. We isolated pre-adipocytes from different adipose depots, omental, subcutaneous and intramuscular, of beef cattle, and cultured in vitro to determine the basis for the variations and attribute these variations to the inherent properties of adipocyte progenitors. The proliferating cells from all depots before the confluence were harvested and the proteome was analyzed by a functional proteomic approach, involving 2-DE and MALDI-TOF/TOF. More than 252 protein spots were identified, selected and analyzed by Image Master (ver 7.0) and MALDI-TOF/TOF. Further, our analysis showed that there were specific differences in proteome expression patterns among proliferating precursor cells from the three depots. Sixteen proteins were found to be differentially expressed and these were identified as proteins involved in cellular processes, heat shock/chaperones, redox proteins, cytoskeletal proteins and metabolic enzymes. The results also enabled us to understand the basic roles of these proteins in different inherent properties exhibited by adipose tissue depots.
Proteomic Analysis of Bovine Longissimus Muscle Satellite Cells during Adipogenic Differentiation
Rajesh, Ramanna Valmiki,Park, Mi-Rim,Heo, Kang-Nyeong,Yoon, Du-Hak,Kim, Tae-Hun,Lee, Hyun-Jeong Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.5
Satellite cells are skeletal muscle progenitor/stem cells that reside between the basal lamina and plasma membranes of skeletal fibers in vivo. These cells can give rise to both myogenic and adipogenic cells. Given the possible role for differentiation of satellite cells into adipocytes in marbling and in some pathological disorders like sarcopenia, knowledge of the proteins involved in such process remains obscure. Using two-dimensional polyacrylamide gel electrophoresis coupled with mass spectrometry, we investigated the proteins that are differentially expressed during adipogenic differentiation of satellite cells from bovine longissimus muscle. Our proteome mapping strategy to identify the differentially expressed intracellular proteins during adipogenic differentiation revealed a total of 25 different proteins. The proteins up-regulated during adipogenic differentiation of satellite cells like Cathepsin H precursor, Retinal dehydrogenase 1, Enoyl-CoA hydratase, Ubiquinol-cytochrome-c reductase, T-complex protein 1 subunit beta and ATP synthase D chain were found to be associated with lipid metabolism. The down-regulated proteins like LIM protein, annexin proteins, cofilin-1, Rho GDP-dissociation inhibitor 1 and septin-2, identified in the present study were found to be associated with myogenesis. These results clearly demonstrate that the adipogenic conversion of muscle satellite cells is associated with the up-regulated and down-regulated proteins involved in adipogenesis and myogenesis respectively.
Enhanced Production and Partial Purification of Glucoamylase from Mutated Bacillus sp. FME
( Gudi Satheesh Kumar ),( Muni Ramanna Gari Subhosh Chandra ),( Yakasiri Nagasai Sujana ),( Bontha Rajasekhar Reddy ),( Yong Lark Choi ) 한국응용생명화학회 2009 Applied Biological Chemistry (Appl Biol Chem) Vol.52 No.5
Enhanced Production and Partial Purification of Glucoamylase from Mutated Bacillus sp. FME
Gudi Satheesh Kumar,Muni Ramanna Gari Subhosh Chandra,Yakasiri Nagasai Sujana,Bontha Rajasekhar Reddy,Yong Lark Choi 한국응용생명화학회 2009 Applied Biological Chemistry (Appl Biol Chem) Vol.52 No.5
In this study, a potent newly-isolated glucoamylase producing actively growing cells of Bacillus sp.FME was subjected to UV irradiation and ethidium bromide (EtBr) mutagenesis. The promising colonies were further screened for glucoamylase production via plate assays and submerged enzyme production at the flask level. Amongst all of the tested colonies, the best mutant, Bacillus sp. FME 2, selected from UV irradiation (20 min) and EtBr (1 mg/mL), was shown to be the most promising. The yield of glucoamylase generated by the mutant strain was approximately 3.0 fold and 1397 U/mL with an incubation period of 24 h, which was larger than the yield generated by the wild-type strain. The glucoamylase was partially purified using ammonium sulphate precipitation followed by gel exclusion chromatography. The enzyme was partially purified 3.0-fold to homogeneity with a final recovery of 66% and a specific activity of 1145 U/mg protein for the mutant strain. The molecular mass was approximately 67.1 kDa, as determined by SDS-PAGE. The active band was observed as a clear colorless area on zymogram analysis, which indicated an absence of glucoamylase isoforms. The results of thin-layer chromatography identified this enzyme as a glucoamylase.
Growth and Foliar Constituents of Mulberry (M5) Cultivated under Organic Based Nutrient Management
Krishnegowda Rashmi,Maruvanahalli Ankegowda Shankar,Kaluvarahalli Ramanna Shashidhar,Talagavara Kempaiah Narayanaswamy 한국잠사학회 2009 International Journal of Industrial Entomology Vol.19 No.1
A field experiment to evaluate the effect of application of different organic manures and inorganic fertilizers on growth, yield and quality of leaf was studied during 2004-05 has showed that, the application of 10 kg each of Azospirillum brasilense and Aspergillus awamori+20% each of recommended N through compost+green manure (Glyricidia maculata)+castor cake+vermicompost+Urea and remaining P and K through fertilizers (T11) has recorded significantly higher leaf yield (250 g/plant and 34.70 tonnes/ha/yr, respectively) with improvement in growth characters as comparedto control. Leaf quality status was also improved in terms of N (3.19%), P (1.97%), K (1.28%), total soluble protein (8.39 mg/ml), total soluble sugars (14.40 mg/ml), secondary nutrients viz., Ca (3.00%), Mg (0.60%), S (0.35%) and micronutrients viz., Cu (0.410 ppm), Mn (0.454 ppm) and Zn (0.112 ppm) contents. The mulberry grown with 20 tonnes of compost+300:120:120 Kg of NPK/ha/year through fertilizer ranked second for growth and foliar constituents.
Jeong, Jin Young,Kim, Jang Mi,Rajesh, Ramanna Valmiki,Suresh, Sekar,Jang, Gul Won,Lee, Kyung-Tai,Kim, Tae Hun,Park, Mina,Jeong, Hak Jae,Kim, Kyung Woon,Cho, Yong Min,Lee, Hyun-Jeong The Korean Society of Animal Reproduction 2013 Reproductive & developmental biology Vol.37 No.4
Satellite cells were derived from muscular tissue in postnatal pig. Satellite cell is an important to growth and development in animal tissues or organs. However, the progress underlying induced differentiation is not clear. The aim of this study was to evaluate the morphologic and the transcriptome changes in porcine satellite cell (PSC) treated with insulin, rosiglitazone, or dexamethasone respectively. PSC was obtained from postnatal muscle tissue. In study 1, for study the effect of insulin and FBS on the differentiated satellite cells, cells were cultured at absence or presence of insulin treated with FBS. Total RNA was extracted for determining the expression levels of myogenic PAX3, PAX7, Myf5, MyoD, and myogenin genes by real-time PCR. Myogenic genes decreased expression levels of mRNA in treated with insulin. In study 2, in order to clarify the relationship between rosiglitazone and lipid in differentiated satellite cells, we further examined the effect of FBS on lipid accumulation in the presence or absence of the rosiglitazone and lipid. Significant differences were observed between rosiglitazone and lipid by FBS. The mRNA of FABP4 and $PPAR{\gamma}$ increased in rosiglitazone treatment. In study 3, we examined the effect of dexamethasone on osteogenic differentiation in PSC. The mRNA was increased osteoblasotgenic ALP and ON genes treated with dexamethasone in 2% FBS. Dexamethasone induces osteoblastogenesis in differentiated PSC. Taken together, in differentiated PSCs, FABP4 and $PPAR{\gamma}$ increased to rosiglitazone. Whereas, no differences to FBS and lipid. These results were not comparable with previous reports. Our results suggest that adipogenic, myogenic, and osteoblastogenic could be isolated from porcine skeletal muscle, and identify culture conditions which optimize proliferation and differentiation formation of PSC.