Adipose tissue in the loin muscle area of beef cattle as a marbling factor is directly associated with beef quality. To elucidate whether properties of proteins involved in depot specific adipose tissue were sex-dependent, we analyzed protein expression of intramuscular adipose tissue (IMAT) and ome...
Adipose tissue in the loin muscle area of beef cattle as a marbling factor is directly associated with beef quality. To elucidate whether properties of proteins involved in depot specific adipose tissue were sex-dependent, we analyzed protein expression of intramuscular adipose tissue (IMAT) and omental adipose tissue (OMAT) from Hanwoo cows, steers, and bulls of Korean native beef cattle by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis, quantitative polymerase chain reaction (PCR) and western blot analysis. Two different adipose depots (i.e. intramuscular and omental) were collected from cows (n = 7), steers (n = 7), or bulls (n = 7). LC-MS/MS revealed a total of 55 and 35 proteins in IMAT and OMAT, respectively. Of the 55 proteins identified, 44, 40, and 42 proteins were confirmed to be differentially expressed in IMAT of cows, steers, and bulls, respectively. In OMAT of cows, steers, and bulls, 33, 33, and 22 were confirmed to be differentially expressed, respectively. Tropomyosin (TPM) 1, TPM 2, and TPM3 were subjected to verification by quantitative PCR and western blot analysis in IMAT and OMAT of Hanwoo cows, steers, and bulls as key factors closely associated with muscle development. Both mRNA levels and protein levels of TPM1, TPM2, and TPM3 in IMAT were lower in bulls compared to in cows or steers suggesting that they were positively correlated with marbling score and quality grade. Our results may aid the regulation of marbling development and improvement of meat quality grades in beef cattle.
Adipose tissue in the loin muscle area of beef cattle as a marbling factor is directly associated with beef quality. To elucidate whether properties of proteins involved in depot specific adipose tissue were sex-dependent, we analyzed protein expression of intramuscular adipose tissue (IMAT) and omental adipose tissue (OMAT) from Hanwoo cows, steers, and bulls of Korean native beef cattle by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis, quantitative polymerase chain reaction (PCR) and western blot analysis. Two different adipose depots (i.e. intramuscular and omental) were collected from cows (n = 7), steers (n = 7), or bulls (n = 7). LC-MS/MS revealed a total of 55 and 35 proteins in IMAT and OMAT, respectively. Of the 55 proteins identified, 44, 40, and 42 proteins were confirmed to be differentially expressed in IMAT of cows, steers, and bulls, respectively. In OMAT of cows, steers, and bulls, 33, 33, and 22 were confirmed to be differentially expressed, respectively. Tropomyosin (TPM) 1, TPM 2, and TPM3 were subjected to verification by quantitative PCR and western blot analysis in IMAT and OMAT of Hanwoo cows, steers, and bulls as key factors closely associated with muscle development. Both mRNA levels and protein levels of TPM1, TPM2, and TPM3 in IMAT were lower in bulls compared to in cows or steers suggesting that they were positively correlated with marbling score and quality grade. Our results may aid the regulation of marbling development and improvement of meat quality grades in beef cattle.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
가설 설정
Therefore, identification of differentially expressed proteins in adipose depots of different sexes might be helpful in defining the functions of intramuscular fat, therefore providing strategies to control meat lipid content independently of body fat depots. The objective of this study was to determine whether proteins involved in depot specific adipose tissue properties were sex dependent. We analyzed the proteome expression of intramuscular adipose tissue (IMAT) and omental adipose tissue (OMAT) in native Hanwoo Korean beef cattle (cows, steers, and bulls) by liquid chromatography-tandem mass spectrometry (LC-MS/MS)–based proteomic analysis, quantitative polymerase chain reaction (PCR), and western blot analysis.
제안 방법
Complete protocol was followed for analytical gels. For preparative gels, the protocol was modified. Glutaraldehyde was omitted from the sensitization step.
To obtain a comprehensive overview of protein components in IMAT and OMAT from individual seven groups (cows, steers, and bulls), protein profiles of whole lysate of IMAT and OMAT were separated by SDS-PAGE and assessed by silver-stained image analysis. The number, marbling score, and quality grade of individuals were showed in Figure 1A.
To study the patterns of gene expression in IMAT and OMAT associated with sex, we used cows, steers, and bulls. The mRNA expression levels of the selected genes were subjected to quantitative real-time PCR with specific primers (Table 1).
대상 데이터
A total of 16 up- or down-regulated proteins between IMAT and OMAT of cows, steers, and bulls were selected. GO analysis was performed using DAVID Bioinformatics Resources 6.
In OMAT of cows, steers, and bulls, 33, 33, and 22 were confirmed to be differentially expressed, respectively (Table 3). All identified proteins were clustered into eight categories based on biological process (BP) using information obtained from the DAVID gene ontology (GO) database (http://david.abcc. ncifcrf.gov) and UniProt (http://www.uniprot.org). Depending on the BP in which the proteins were involved, they were categorized into the following groups (Figure 2A): carbohydrate metabolism (35.
We used adipose tissue samples of cows (n = 7), steers (n =7), and bulls (n = 7). Tissue samples were collected in three animals groups from two different adipose depots (i.e. intramuscular and omental). Slaughter age was approximately 31 months for all cattles.
데이터처리
Statistical significance was evaluated using Student’s t-test.
Student’s t test was performed to evaluate statistical significance (*** p<0.0001; mean±standard error of the mean; n = 3).
이론/모형
Relative quantification analysis was performed using the comparative Ct (2–ΔΔCt) method (Wilting et al., 2010).
성능/효과
0001) lower expression in IMAT than in OMAT in bulls (Figure 3C). These results demonstrated that ALDOA was differentially expressed depending on sex, suggesting that ALDOA could be one of the factors affecting lipid accumulation in OMAT.
참고문헌 (36)
Arnold AM Peralta JM Thonney ML 1996 Ontogeny of growth hormone, insulin-like growth factor-I, estradiol and cortisol in the growing lamb: Effect of testosterone J Endocrinol 150 391 399 8882158
Choy YH Park BH Choi TJ Choi JG Cho KH Lee SS Choi YL Koh KC Kim HS 2012 Estimation of relative economic weights of hanwoo carcass traits based on carcass market price Asian Australas J Anim Sci 25 1667 1673 25049531
Colbert MC Ciejek-Baez E 1988 Alternative promoter usage by aldolase A during in vitro myogenesis Dev Biol 130 392 396 3181637
Destefanis G Brugiapaglia A Barge MT Lazzaroni C 2003 Effect of castration on meat quality in Piemontese cattle Meat Sci 64 215 218 22062869
Dlugosz AA Antin PB Nachmias VT Holtzer H 1984 The relationship between stress fiber-like structures and nascent myofibrils in cultured cardiac myocytes J Cell Biol 99 2268 2278 6438115
Enns DL Iqbal S Tiidus PM 2008 Oestrogen receptors mediate oestrogen-induced increases in post-exercise rat skeletal muscle satellite cells Acta Physiol (Oxf) 194 81 93 18397384
Gondret F Guitton N Guillerm-Regost C Louveau I 2008 Regional differences in porcine adipocytes isolated from skeletal muscle and adipose tissues as identified by a proteomic approach J Anim Sci 86 2115 2125 18310487
Guo B Kongsuwan K Greenwood PL Zhou G Zhang W Dalrymple BP 2014 A gene expression estimator of intramuscular fat percentage for use in both cattle and sheep J Anim Sci Biotechnol 5 35 25028604
Hidaka K Yamamoto I Arai Y Mukai T 1993 The MEF-3 motif is required for MEF-2-mediated skeletal muscle-specific induction of the rat aldolase A gene Mol Cell Biol 13 6469 6478 8413246
Hughes JM Oiseth SK Purslow PP Warner RD 2014 A structural approach to understanding the interactions between colour, water-holding capacity and tenderness Meat Sci 98 520 532 25034451
Inoue K Yamasaki S Fushiki T Okada Y Sugimoto E 1994 Androgen receptor antagonist suppresses exercise-induced hypertrophy of skeletal muscle Eur J Appl Physiol Occup Physiol 69 88 91 7957162
Jeong J Bong J Kim GD Joo ST Lee HJ Baik M 2013 Transcriptome changes favoring intramuscular fat deposition in the longissimus muscle following castration of bulls J Anim Sci 91 4692 4704 23893995
Jeong J Kwon EG Im SK Seo KS Baik M 2012 Expression of fat deposition and fat removal genes is associated with intramuscular fat content in longissimus dorsi muscle of Korean cattle steers J Anim Sci 90 2044 2053 22266990
Kahlert S Grohe C Karas RH Lobbert K Neyses L Vetter H 1997 Effects of estrogen on skeletal myoblast growth Biochem Biophys Res Commun 232 373 378 9125184
Lee DM Bajracharya P Lee EJ Kim JE Lee HJ Chun T Kim J Cho KH Chang J Hong S Choi I 2011 Effects of gender-specific adult bovine serum on myogenic satellite cell proliferation, differentiation and lipid accumulation In Vitro Cell Dev. Biol. Anim. 47 438 444
Lee SH Park EW Cho YM Kim SK Lee JH Jeon JT Lee CS Im SK Oh SJ Thompson JM Yoon D 2007 Identification of differentially expressed genes related to intramuscular fat development in the early and late fattening stages of hanwoo steers J Biochem Mol Biol 40 757 764 17927910
Lin JJ Lin JL 1986 Assembly of different isoforms of actin and tropomyosin into the skeletal tropomyosin-enriched microfilaments during differentiation of muscle cells in vitro J Cell Biol 103 2173 2183 3536961
Maltin C Balcerzak D Tilley R Delday M 2003 Determinants of meat quality: tenderness Proc Nutr Soc 62 337 347 14506881
Marston S Memo M Messer A Papadaki M Nowak K McNamara E Ong R El-Mezgueldi M Li X Lehman W 2013 Mutations in repeating structural motifs of tropomyosin cause gain of function in skeletal muscle myopathy patients Hum Mol Genet 22 4978 4987 23886664
Nishimura T 2010 The role of intramuscular connective tissue in meat texture Anim Sci J 81 21 27 20163668
Park GB Moon SS Ko YD Ha JK Lee JG Chang HH Joo ST 2002 Influence of slaughter weight and sex on yield and quality grades of Hanwoo (Korean native cattle) carcasses J Anim Sci 80 129 136 11831510
Peachey BM Purchas RW Duizer LM 2002 Relationships between sensory and objective measures of meat tenderness of beef m. longissimus thoracis from bulls and steers Meat Sci 60 211 218 22063391
Purchas RW Burnham DL Morris ST 2002 Effects of growth potential and growth path on tenderness of beef longissimus muscle from bulls and steers J Anim Sci 80 3211 3221 12542162
Purslow PP 2005 Intramuscular connective tissue and its role in meat quality Meat Sci 70 435 447 22063743
Ren H Li L Su H Xu L Wei C Zhang L Li H Liu W Du L 2011 Histological and transcriptome-wide level characteristics of fetal myofiber hyperplasia during the second half of gestation in Texel and Ujumqin sheep BMC Genomics 12 411 21838923
Schreurs NM Garcia F Jurie C Agabriel J Micol D Bauchart D Listrat A Picard B 2008 Meta-analysis of the effect of animal maturity on muscle characteristics in different muscles, breeds, and sexes of cattle J Anim Sci 86 2872 2887 18599671
Sinha-Hikim I Roth SM Lee MI Bhasin S 2003 Testosterone-induced muscle hypertrophy is associated with an increase in satellite cell number in healthy, young men Am J Physiol Endocrinol Metab 285 E197 205 12670837
Walsh TP Winzor DJ Clarke FM Masters CJ Morton DJ 1980 Binding of aldolase to actin-containing filaments. Evidence of interaction with the regulatory proteins of skeletal muscle Biochem J 186 89 98 6892770
Walter LJ Gasch CA McEvers TJ Hutcheson JP Defoor P Marquess FL Lawrence TE 2014 Association of pro-melanin concentrating hormone genotype with beef carcass quality and yield J Anim Sci 92 325 331 24243910
Wilting SM van Boerdonk RA Henken FE Meijer CJ Diosdado B Meijer GA le Sage C Agami R Snijders PJ Steenbergen RD 2010 Methylation-mediated silencing and tumour suppressive function of hsa-miR-124 in cervical cancer Mol Cancer 9 167 20579385
Yan JX Wait R Berkelman T Harry RA Westbrook JA Wheeler CH Dunn MJ 2000 A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization-mass spectrometry Electrophoresis 21 3666 3672 11271485
Zhang HM Su YF Shi ZY Fu YS 2014 cDNA clone and expression analysis of alpha-Tropomyosin during Japanese flounder ( Paralichthys olivaceus ) metamorphosis Dongwuxue Yanjiu 35 307 312 25017750
Zuo X Echan L Hembach P Tang HY Speicher KD Santoli D Speicher DW 2001 Towards global analysis of mammalian proteomes using sample prefractionation prior to narrow pH range two-dimensional gels and using one-dimensional gels for insoluble and large proteins Electrophoresis 22 1603 1615 11425216
※ AI-Helper는 부적절한 답변을 할 수 있습니다.