Microarray analysis of Longissimus thoracis muscle gene expressions in vitamin A‐restricted Japanese Black steers in middle fattening stage

Vitamin A (VA) restriction in beef cattle improves meat marbling; however, the underlying molecular mechanisms remain incompletely understood. We performed microarray analysis to clarify the effect of VA restriction on Longissimus thoracis gene expressions in Japanese Black steers. Six Japanese Black steers 13–14 months of age were divided into two groups: S group (n = 3), which received VA supplementation, and R group (n = 3), in which dietary VA intake was restricted. Steers were fattened for 7 months, following which tissue samples were obtained. Extracted RNA samples were analyzed by Affymetrix Genechip Bovine Genome Array. Lists of genes highly expressed in the R and S groups were obtained. The lists were functionally interpreted using functional annotation software, DAVID. In the R and S groups, 48 and 40 genes were significantly highly expressed, respectively. The gene list of the R group included CD36, LPL, GPAM, DGAT2, and SCD and additional genes annotated ‘PPAR signaling pathway,’ ‘lipid biosynthesis’ and ‘mitochondrion,’ whereas that of the S group included COL1A2, FN1 and DCN and additional genes annotated ‘extracellular matrix.’ Changes in the expression of these genes are possibly involved in marbling improvement in beef cattle by VA restriction.     

维生素A和维生素D对动物基因表达的影响

本文在介绍维生素A和维生素D营养生理功能的基础上,综述了维生素A和维生素D对相关基因表达的调控及其主要机制。     

Relationships among muscle fiber type composition,fiber diameter and MRF gene expression in different skeletal muscles of naturally grazing Wuzhumuqin sheep during postnatal development

The aim of this study was to determine the relationships among muscle fiber‐type composition, fiber diameter, and myogenic regulatory factor (MRF) gene expression in different skeletal muscles during development in naturally grazing Wuzhumuqin sheep. Three major muscles (i.e. the Longissimus dorsi (LD), Biceps femoris (BF) and Triceps brachii (TB)) were obtained from 20 Wuzhumuqin sheep and 20 castrated rams at each of the following ages: 1, 3, 6, 9, 12 and 18 months. Muscle fiber‐type composition and fiber diameter were measured using histochemistry and morphological analysis, and MRF gene expression levels were determined using real‐time PCR. In the LD muscle, changes in the proportion of each of different types of fiber (I, IIA and IIB) were relatively small. In the BF muscle, a higher proportion of type I and a 6.19‐fold lower proportion of type IIA fibers were observed (P < 0.05). In addition, the compositions of type I and IIA fibers continuously changed in the TB muscle (P < 0.05). Moreover, muscle diameter gradually increased throughout development (P < 0.05). Almost no significant difference was found in MRF gene expression patterns, which appeared to be relatively stable. These results suggest that changes in fiber‐type composition and increases in fiber size may be mutually interacting processes during muscle development.     

维生素E、K对动物基因表达的影响

本文在介绍维生素E和维生素K营养生理功能的基础上,综述了维生素E和维生素K对相关基因表达的调控及其主要机制。     

维生素A对动物基因表达的调控

本文主要综述了维生素A对调节视力的视蛋白基因的表达调控,时糖元异生起关键作用磷酸烯醇式丙酮酸羧激酶(PEPCK)基因的调控,多不饱和脂肪酸合成中△5-去饱和酶基因以及对与生长有关的类胰岛素生长因子(IGF)基因的调控,为研究日粮中维生素A添加对动物营养机理的研究提供理论依据.     

Influence of rice whole‐crop silage diet on growth performance,carcass and meat characteristics and muscle‐related gene expression in Japanese Black steers

The present study investigated the influence of a diet largely comprising rice whole‐crop silage (rWCS) on growth performance, carcass and meat characteristics, and expression of genes involved in muscle growth of Japanese Black steers. Steers were randomly separated into rWCS‐fed (rWCS ad libitum and restricted feeding of concentrate) and concentrate‐fed groups. Total digestible nutrient intake and daily gain (DG) decreased in rWCS‐fed steers in comparison with concentrate‐fed steers, whereas dressed carcass weight and final body weight did not significantly differ between the groups. Decreases in drip loss in the muscle of rWCS‐fed steers may be caused by α‐tocopherol and β‐carotene in muscle. Feeding large amounts of rWCS to steers may maintain quantitative productivity of beef steers equally to a concentrate‐based diet, and improve the qualitative productivity. Results of gene expression suggest that activation of skeletal muscle growth in rWCS‐fed steers may occur at the late fattening period owing to a decrease in myostatin and increase in myosin heavy chain gene expression. Preadipocyte factor‐1 and myostatin genes may be strongly involved in the control of lipid accumulation. This rearing system would allow beef production to switch to rWCS‐based diets from concentrate‐based diets.     

日粮维生素A水平对肉鸡组织CaBP基因表达影响的研究

采用完全随机试验设计,将192只8日龄健康AA雄性肉鸡称重后随机分为4个处理组,每处理组6个重复,每个重复8只。在以玉米和豆粕为主要原料的基础日粮中分别添加VA1500,3000,15000和45000U/kg构成试验日粮,试验期5周。应用SYBR GreenI实时定量RT-PCR技术测定肉鸡胫骨、十二指肠组织中CaBP基因表达量,研究不同VA水平对肉鸡组织CaBP基因表达的影响。结果表明:随着日粮VA添加量的增加,十二指肠CaBP基因表达降低,当VA水平增加至15000,45000U/kg时,CaBP基因表达量有降低趋势(P>0.05),其中以45000U/kg添加组的CaBP基因表达量最低。虽然日粮VA水平对肉鸡胫骨组织中CaBP基因的表达影响较小,但当添加水平为45000U/kg时胫骨组织中CaBP基因表达量有降低的趋势。说明过量VA对骨骼代谢影响的机制之一是过量VA降低了与钙、磷代谢有关的CaBP的基因表达,进而影响钙、磷代谢和骨骼发育。     

Correlation between skeletal muscle fiber type and free amino acid levels in Japanese Black steers

Free amino acids are important components of tastants and flavor precursors in meat. To clarify the correlation between muscle fiber type and free amino acids, we determined the concentrations of various free amino acids and dipeptides in samples of different muscle tissues (n = 21), collected from 26‐month‐old Japanese Black steers (n = 3) at 2 days postmortem. The proportions of the myosin heavy chain (MyHC), slow (MyHC1) and fast (MyHC2) isoforms were determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The contents of free amino acids and dipeptides were measured by high performance liquid chromatography (HPLC). The MyHC isoform composition varied among the tissue samples. The MyHC1 proportion ranged from 6.9% ± 3.9% to 83.3% ± 16.7%. We confirmed that there was a strong positive correlation between MyHC1 composition and total free amino acid concentrations, including those for two dipeptides. Among the 31 measured free amino acids and dipeptides, 11 showed significant positive correlations and five showed significant negative correlations with MyHC1 composition. These results suggest that a high MyHC1 content induces high free amino acid contents in bovine muscles possibly because of greater oxidative metabolism. This high level of free amino acids could contribute to the intense flavor of meat that is rich in slow‐twitch fibers.     

Changes in muscle fiber type and expression of mRNA of myosin heavy chain isoforms in porcine muscle during pre‐ and postnatal development

The purpose of this study is to elucidate developmental changes in muscle fiber type in the pig during pre‐ and postnatal development. For this purpose, we performed a histochemical analysis for myosin adenosine triphosphatase activity to assess muscle fiber type and determined abundances of messenger RNA (mRNA) of myosin heavy chain (MHC) isoforms. Samples of Longissimus dorsi (LD) muscle were taken from fetuses on day 90 of the fetal stage. Further, samples of LD, Rhomboideus and Biceps femoris (B. femoris) muscles were taken from pigs when they were 1, 12, 26, 45 or 75 days old. Expression of MHC 2b mRNA in the LD and the B. femoris muscles rapidly and considerably increased from the late fetal stage to the early postnatal stage and this increase was associated with the development of type 2b fibers at least in the LD muscle. As shown by the rapid and considerable changes in expression of MHC 2b mRNA, it seems that a certain plasticity of muscle fiber type still remains in this developmental stage.     

Blood and milk status of vitamin E,vitamin A,and selenium in nursing Awassi ewes injected with vitamin E and selenium

Abstract

This study investigated the effects of vitamin E and selenium (Se) injections on concentrations of vitamin E, vitamin A, and Se in blood and milk to predict when injections may improve ewes’ performance. Awassi ewes received 0 (control, 13 ewes) or 15 plus 0.05?mg/kg BW of vitamin E and Se (injected, 15 ewes) at 1 and 4 weeks postpartum. Blood and milk samples were collected weekly for 6 weeks. Injections increased vitamin E, decreased Se, and had no effects on vitamin A levels in blood. Blood vitamin E was correlated positively with vitamin A. Injections increased vitamin E, but had no effects on vitamin A or Se levels in milk. Milk vitamin A was positively correlated with vitamin E and Se levels in the control group. Milk vitamins E and A were positively correlated with their corresponding levels in blood. Milk somatic cell count was negatively correlated with blood vitamin A levels.     

VA对基因表达的调控

随着分子生物学技术和方法的快速发展及其在营养学中的应用,营养素与基因表达的相互作用已成为研究的热点。在介绍VA营养生理功能的基础上,综述了VA对相关基因表达的调控及其主要机制。     

Effect of supplemental β‐carotene compared to retinyl palmitate on fatty acid profile and expression of mRNA from genes involved in vitamin A metabolism in beef feedlot cattle

To examine the effects of dietary β‐carotene (βC) or retinyl palmitate (RP) on fatty acid (FA) profile and mRNA expression, samples were collected from 24 Angus‐cross calves that were allotted to four treatments consisting of RP supplemented at 2200 IU/kg, and synthetic β‐carotene (SβC) supplemented at one, five or 10 times RP. Longissimus muscle (LM) cis‐9, trans‐11 conjugated linoleic acid was greater in RP compared to SβC1X (P = 0.04). The polyunsaturated:saturated FA increased linearly (P = 0.04) in the LM as dietary SβC increased. Expression of βC oxygenase 2 (βCO2), an enzyme that cleaves β‐carotene, was greater in the LM for SβC1X compared to RP and decreased linearly as SβC increased (P ≤ 0.02). Peroxisome proliferator activated receptor γ (PPARγ) expression in the LM increased in SβC1X compared to RP (P = 0.03); however, PPARγ and retinoic acid X receptor α (RXRα) expression decreased linearly (P = 0.02) in the LM with increasing SβC. Retinoic acid receptor α (RARα) expression tended (P = 0.10) to decrease linearly in the LM with increased SβC. In conclusion, SβC supplementation increased mRNA expression of some lipogenic genes in the LM, but increasing dietary SβC inhibited their expression and tended to increase polyunsaturated FA.     

Grazing‐induced changes in muscle microRNA‐206 and ‐208b expression in association with myogenic gene expression in cattle

To investigate the roles of microRNAs (miRNAs) in muscle type conversion, the effects of 4 months of grazing on the expression levels of miRNAs and mRNAs associated with skeletal muscle development were analyzed by quantitative RT‐PCR using the Biceps femoris muscle of Japanese Shorthorn cattle. After 4 months of grazing, the expression of muscle fiber type‐associated miR‐208b was higher in the grazed cattle than in the housed. In concordance with the pattern in miR‐208b expression, the expression of MyoD, a myogenic regulatory factor associated with the shifting of muscle property to the fast type, was lower in the grazed cattle after 4 months of grazing than in the housed cattle. In addition, the expression of MyHC‐2x (a fast type) was higher in the housed cattle than in the grazed, after 4 months of grazing. During the grazing period, miR‐206 expression decreased in the housed cattle, whereas expression in the grazed cattle did not change, but rather remained higher than that of the housed cattle even at 3 months after the grazing ended. These miRNAs including miR‐206 persisting with muscles of grazed cattle may be associated with regulation of muscle gene expression during skeletal muscle adaptation to grazing.     

测定饲料中维生素A和维生素E含量方法的研究

本文旨在建立一种能同时准确测定饲料中维生素A和维生素E含量的方法,试验将样品在碱性条件下65℃水浴超声皂化提取,冰乙酸调pH至中性,乙醇稀释后直接用高效液相色谱法(HPLC)进行测定。结果表明:维生素A在0.036~108IU/mL线性关系良好,相关系数为1.00000,检出限1000IU/kg,方法回收率为95.8%~98.8%,RSD为0.8%~4.4%(n=5);维生素E在0.6~125μg/mL线性关系良好,相关系数为0.99998,检出限15mg/kg,方法回收率为95.3%~102.2%,RSD为0.8%~4.2%(n=5)。本方法快速简便,适用于配合料、浓缩料、维生素预混料和复合预混料中同时准确测定维生素A和维生素E的含量,相对于国标法节省了大量的时间和试剂,可应用于日常大批量饲料检测中。     

Effects of starch and gelatin encapsulated vitamin A on growth performance,immune status and antioxidant capacity in weaned piglets

To evaluate the effects of gelatin and starch encapsulated vitamin A on growth performance, immune status and antioxidant capacity in weaned piglets, a total of 96 weaned piglets (body weight = 9.11 ± 0.03 kg, 30-d-old) were randomly allotted to 3 treatments with 4 replications of 8 piglets each. The 3 treatments were control diet (basal diet without addition of vitamin A), gelatin vitamin A diet (basal diet + 13,500 IU/kg gelatin encapsulated vitamin A), and starch vitamin A diet (basal diet + 13,500 IU/kg starch encapsulated vitamin A), respectively. The results showed that piglets fed starch vitamin A diet had significantly higher final body weight and average daily gain compared to those in control and gelatin vitamin A groups (P < 0.05). Gelatin and starch vitamin A supplementation both highly increased serum retinol concentration and immunoglobulin (Ig) M level when compared with the control group (P < 0.05). Additionally, serum IgA level and glutathione peroxidase (GSH-Px) activity were significantly increased by gelatin vitamin A diet on d 21 and starch vitamin A diet on d 42, respectively (P < 0.05). These results demonstrated that dietary supplementation of vitamin A could improve immune function and antioxidant capacity in weaned piglets, and starch vitamin A is better than gelatin vitamin A, especially in promoting the growth performance of piglets.     

MyoD and Myf6 gene expression patterns in skeletal muscle during embryonic and posthatch development in the domestic duck (Anas platyrhynchos domestica)

The MyoD and Myf6 genes, which are muscle regulatory factors (MRFs), play major roles in muscle growth and development and initiate muscle fibre formation via the regulation of muscle‐specific gene translation. Therefore, MyoD and Myf6 are potential candidate genes for meat production traits in animals and poultry. The objective of this study was to evaluate MyoD and Myf6 gene expression patterns in the skeletal muscle during early developmental stage of ducks. Gene expression levels were detected using the quantitative RT‐PCR method in the breast muscle (BM) and leg muscle (LM) at embryonic days 13, 17, 21, 25, 27, as well as at 1 week posthatching in Gaoyou and Jinding ducks (Anas platyrhynchos domestica). The MyoD and Myf6 gene profiles in the two duck breeds were consistent during early development, and MyoD gene expression showed a ‘wave’ trend in BM and an approximate ‘anti‐√’ trend in LM. Myf6 gene expression in BM showed the highest level at embryonic day 21, which subsequently decreased, although remained relatively high, while levels at embryonic days 13, 17 and 21 were higher in LM. The results of correlation analysis showed that MyoD and Myf6 gene expression levels were more strongly correlated in LM than in BM in both duck breeds. These results indicated that different expression patterns of the MyoD and Myf6 genes in BM and LM may be related to muscle development and differentiation, suggesting that MyoD and Myf6 are integral to skeletal muscle development.     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

Muscle structure and gene expression in pectoralis major muscle in response to deep pectoral myopathy induction in fast- and slow-growing commercial broilers

1. The objective of this study was to determine muscle structure and gene expression in pectoralis major (p. major) muscle of broilers in response to deep pectoral myopathy (DPM) induction.

2. A total of 160 chickens from slow- and fast-growing broilers were raised under same conditions. Half of the broilers from each strain were encouraged to wing flap when they reached 2800 g body weight. Pectoralis minor (p. minor) muscle of the broilers was inspected for the occurrence of DPM and p. major samples were collected from broilers with or without DPM. The muscle fibre area and number, capillary number and the signalling pathways of vascular development (vascular endothelial growth factor A, VEGFA) and muscle contraction regulation (actin alpha 1, ACTA1; myosin light chain kinase 2, MYLK2 and ATPase Ca⁺² transporting gene 1, ATP2A1) were studied in p. major muscle.

3. DPM induction increased fibre area of p. major muscle with a greater rate in the slow-growing strain compared with fast-growing line. Although the capillary number was higher in slow-growing compared with fast-growing broilers, in the case of DPM induction, the number of capillaries was similar between strains.

4. Expression of VEGFA, MYLK2 and ATP2A1 was greater in slow- than in fast-growing broilers. DPM induction increased expression of ACTA1, VEGFA and ATP2A1 in p. major muscle of broilers from both strains; however, MYLK2 expression was downregulated.

5. Changes in capillary density and expression of VEGFA found in the p. major muscle of broilers with DPM suggest increased blood flow to increase oxygen availability. The upregulation of ATP2A1 by DPM induction could be attributable to alterations in calcium ion transportation from the cytoplasm into the sarcoplasmic reticulum.

6. The results are evidence of changes in muscle structure and gene expression pathways in p. major muscle of broilers with DPM.