不同品种猪肌纤维组成成分及相关信号通路基因的表达研究

发布时间：2017-12-30 18:02

本文关键词：不同品种猪肌纤维组成成分及相关信号通路基因的表达研究　出处：《锦州医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的探索不同品种猪出生后肌纤维组成成分及相关信号通路关键成分的基因表达规律及种间差异,同时分析猪肉质性状候选基因CFL2与肌纤维组成成分和信号通路关键成分的相关性,为建立猪肌纤维性状的检测方法提供前期工作基础。方法选择饲养条件相同、同一父系的纯种荷包猪、大白猪、长白猪和杜长大三元杂交商品猪,按常规方法分别于1d、30d、60d、90d、120d、150d和180d日龄屠宰母猪各3头,共计84头。屠宰时测定肉质性状(肌纤维直径、剪切力、肉色和p H)。屠宰后背最长肌的肌肉样品,采用实时定量PCR和Western blot技术对肌纤维组成成分(My HCslow、My HC2a、My HC2x、My HC2b、MLC、α-actin)、信号通路关键成分(MEF2C、sk MLCK、Rho A、AMPKα2、MAPKp38及Ca M)及肌纤维候选基因CFL2的m RNA和蛋白质表达情况进行检测;并对检测数据采用SPSS17.0统计软件进行多重比较方差分析。结果1、大白和长白猪的肉质性状变化基本一致。随着猪的生长发育,荷包、大白和长白猪的肉色显著加深、剪切力显著增加、肌纤维直径显著增大(P0.05);不同猪种的肉色和p H在120d之后差异不显著;180d时,各品种猪剪切力差异不显著,而荷包猪的肌纤维直径与大白、长白差异显著(P0.05)。2、各基因的m RNA检测发现:同一基因在不同猪种中有大致相同的生后表达规律,但不同品种相同时间点存在表达量的显著差异。大白和长白猪的My HC总量和肌纤维组分的m RNA表达量基本一致,而荷包猪60d-150d与大白、长白猪差异显著(P0.05);不同品种猪CFL2 m RNA表达趋势基本与My HC总量和肌纤维组分表达相似。1d-60d不同品种MEF2C m RNA表达差异不显著,90d-150d荷包猪显著高于其他品种,180d荷包猪显著低于其他品种。sk MLCK m RNA表达量总体呈上升趋势;AMPK和MAPKp38 m RNA表达量却随日龄增长下降。长白猪30d-180d的Rho A m RNA表达始终极显著高于其他猪种(P0.01);荷包猪90d-180d的CAM m RNA表达都极显著高于其他品种(P0.01)。3、利用Western Blot方法检测蛋白表达,结果显示:各品种猪的蛋白表达趋势基本一致,差异在于表达量不同。荷包猪My HC slow总体极显著(P0.01)高于其它品种;其他的My HC总量和肌纤维成分(My HC2a、My HC2x、My HC2b、MLC和α-actin),除个别时间点差异不显著,长白猪总体显著高于其它品种(P0.05)。荷包猪的CFL2蛋白表达量自出生至180d始终极显著高于进口猪种(大白、长白和三元杂交猪)(P0.01)。长白猪sk MLCK蛋白表达始终极显著高于其他品种(P0.01),而荷包猪与大白猪相似且最低。随猪日龄增长,AMPK表达量下降,而MAPKp38整体升高。Rho A蛋白,长白猪1d-150d始终极显著高于其他品种(P0.01),仅180d低于荷包猪。荷包猪与三元猪的CAM表达量相似,高于大白和长白猪。4、相关分析表明:所有肉质性状均与日龄极显著相关(P0.01);肌纤维组成成分中,仅My HC2a与日龄极显著(P0.01)正相关,α-actin与日龄显著(P0.05)负相关;信号通路AMPKα2和MAPKp38与日龄呈中等程度显著(P0.05)相关。My HC2b和My HC2a分别与p H24和肌纤维直径显著相关(P0.05);信号通路蛋白中AMPKα2与所有肉质性状有关,MAPKp38和Ca M仅与剪切力和肌纤维直径中、低度相关。CFL2的表达仅与My HC slow呈极显著(P0.01)正相关;My HC总量与MLC也呈极显著(P0.01)正相关。除Ca M外,信号通路关键成分(MEF2C、sk MLCK、AMPKα2、MAPKp38、Rho A)均与肌纤维成分中、低度相关。结论1猪的肉质性状和肌球蛋白My HC2a表达与日龄显著正相关。2肌球蛋白My HC2b的含量影响肉质的p H,My HC2a的含量影响肌纤维直径。中国地方猪—荷包猪肉质好于进口猪种的原因之一是肌纤维直径小。3信号通路AMPKα2与所有肉质性状相关,MAPKp38和Ca M仅与剪切力和肌纤维直径中低度相关。4 CFL2可能通过调节My HC slow来影响肌纤维组成,促进肌纤维类型改变进而参与影响肉质性状。
[Abstract]:Objective to explore the expression of different varieties of pigs after the birth of muscle fiber composition, key components and related signal pathway genes and interspecific difference, correlation analysis of pork quality traits candidate gene CFL2 and muscle fiber composition key components of signaling pathways and at the same time, provide a basis to establish a detection method of pig muscle fiber traits. The choice of feeding method under the same conditions, the same male purebred Hebao pigs, large white, Landrace and Duroc crossbred pigs three yuan, according to the conventional method respectively in 1D, 30d, 60d, 90d, 120d, 150D and 180d day old mother pig slaughter each 3, a total of 84 head. Determination of meat quality traits at slaughter (diameter of muscle fiber shear force, color and P, H). Slaughter back and the longest muscle samples by real-time quantitative PCR and Western blot technology on muscle fiber composition (My HCslow, My HC2a, My HC2x, My HC2b, MLC, -actin, alpha) signal pathway Components (MEF2C, SK MLCK, Rho A, AMPK MAPKp38 and Ca alpha 2, M) was used to detect the expression of M RNA and protein and muscle fiber candidate gene CFL2; and the test data for multiple comparisons ANOVA using SPSS17.0 statistical software. Results 1 Changes of meat quality traits in large white and Landrace with basically the same. The growth of pigs, large white and Landrace pouch, color significantly deepened, shear stress significantly increased, muscle fiber diameter increased significantly (P0.05); different pig breeds and P H color after 120d 180d, the difference was not significant; the pig breeds shear stress difference is not significant, and Hebao pig muscle fiber the diameter and large white, Landrace was significantly different (P0.05).2, found m RNA to detect the gene, the same gene is roughly the same in different pig breeds after expression pattern, but different varieties at the same time there was significant difference. The amount of large white and Landrace My and total HC M RNA muscle fiber component expression is basically the same, and Hebao pig 60d-150d with large white, Landrace was significantly different (P0.05); different kinds of expression of porcine CFL2 and My HC m RNA the basic trend and total muscle fiber composition similar to different varieties of.1d-60d MEF2C m expression RNA expression had no significant difference, 90d-150d was significantly higher than that of other Hebao pigs 180d varieties, Hebao pigs was significantly lower than the other varieties of.Sk MLCK m RNA expression in the overall upward trend; AMPK MAPKp38 and m RNA expression with age decreased. 30d-180d Rho A m RNA of Landrace was significantly higher than that of the ultimate expression of other breeds (P0.01); the expression of CAM m RNA 90d-180d are significant Hebao pigs higher than other varieties (P0.01).3 expression, using Western method for the detection of Blot protein results showed that all varieties of pig protein expression trend was consistent. The difference lies in the difference in expression of My HC slow overall Hebao pigs (P0.01) were significantly higher In other varieties; the other My HC and total muscle fiber composition (My HC2a, My HC2x, My HC2b, MLC and alpha -actin), in addition to the individual time points were not significantly different, Landrace overall was significantly higher than that of other varieties (P0.05). The expression of CFL2 protein of Hebao pigs from birth to 180d was significantly higher than that of the ultimate import pig breeds (large white, Landrace and three crossbred pigs) (P0.01). The expression of SK MLCK protein was the ultimate Landrace was significantly higher than that of other varieties (P0.01), and the purse and large white pigs. Pigs with similar and minimum age, lower expression of AMPK, MAPKp38 and.Rho increased the overall A protein was 1d-150d, Landrace the ultimate was significantly higher than that of other varieties (P0.01), only 180d is lower than that of Hebao pig Hebao pig and pig three yuan. The expression of CAM was similar to that of large white and Landrace pigs was higher than.4, the correlation analysis showed that: all meat traits were significantly correlated with age (P0.01); muscle fiber components, only My HC2a and age pole Significant positive correlation (P0.01), alpha -actin (P0.05) and age was negatively correlated; moderately significant signal pathway of AMPK alpha 2 and MAPKp38 with age (P0.05) and My HC2b.My correlated HC2a respectively with P H24 and muscle fiber diameter (P0.05); AMPK signaling pathway protein 2 and alpha all MAPKp38 and Ca on meat quality traits, only M and shear force and muscle fiber diameter, low expression of related.CFL2 only with My HC slow (P0.01) showed a significant positive correlation; the total HC and MLC My also showed significant positive correlation (P0.01). In addition to Ca M, a key component of signal pathway (MEF2C SK, MLCK, AMPK, MAPKp38, Rho, alpha 2, A) were related to low and muscle fiber composition, expression. Conclusion 1 pig meat quality traits and myosin My HC2a and age was positively related to.2 My HC2b the effects of the content of myosin P H influence the content of meat, muscle fiber diameter. My HC2a in our local pig - fried pork in good One of the reasons is the kind of pig muscle fiber diameter small.3 signaling pathway AMPK alpha 2 associated with all meat quality traits, MAPKp38 and M only Ca associated with low shear stress and muscle fiber diameter in.4 CFL2 My HC slow may regulate the effect of muscle fibers, promote muscle fiber type and is involved in the change influencing meat quality traits.

【学位授予单位】：锦州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S828

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