Growth factors improve the proliferation of Jeju black pig muscle cells by regulating myogenic differentiation 1 and growth-related genes

Park, Jinryong,Lee, Jeongeun,Song, Ki-Duk,Kim, Sung-Jo,Kim, Dae Cheol,Lee, Sang Cheol,Son, Young June,Choi, Hyun Woo,Shim, Kwanseob 아세아태평양축산학회 2021 Animal Bioscience Vol.34 No.8

Objective: The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells. Methods: Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5×10⁵ cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (MyoD) expression and growth-related gene expression in JBP muscle cells, respectively. Results: We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (EGF+bFGF) than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of collagen type XXI alpha 1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells. Conclusion: Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.

Induction of Ski Protein Expression upon Luteinization in Rat Granulosa Cells

Kim, Hyun,Kim, Dong-Hun,Park, Soo-Bong,Ko, Yeoung-Gyu,Kim, Sung-Woo,Do, Yoon-Jun,Park, Jae-Hong,Yang, Boh-Suk 아세아태평양축산학회 2012 Animal Bioscience Vol.25 No.5

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells; however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to locate Ski protein in the rat ovary during luteinizationto predict the possible role of Ski. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rats, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of the corpus luteum (CL). Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggests that its expression is regulated post-transcriptionally.

Effect of Buffalo Follicular Fluid Alone and in Combination with PMSG and M199 on in vitro Buffalo Oocyte Maturation

Gupta, P.S.P.,Nandi, S.,Ravindranatha, B.M.,Sarma, P.V. 아세아태평양축산학회 2001 Animal Bioscience Vol.14 No.5

The effect of replacement of in vitro maturation medium completely with the buffalo follicular fluid (buFF) on in vitro oocyte maturation of buffalo oocytes was studied. 5 to 8 buffalo cumulus oocyte complexes were cultured in a single drop with each of the eight media studied i.e., M199+steer serum (10% v/v), M199+steer serum (10% v/v)+PMSG, M199+buFF (10% v/v), M199+buFF (10% v/v)+PMSG, M199+buFF (50% v/v), M199+buFF (50% v/v)+ PMSG, buFF (100%) and buFF+PMSG at $39^{\circ}C$ and 5% $CO_2$ in air for 24 h. Supplementation of M199 with Steer serum alone resulted in IVM rate of 35% only. When the above medium was supplemented with PMSG, the maturation rate rallied to 82%. Significant increase in the maturation rates were observed when M199 was supplemented with increasing levels of buFF. A further increase in the maturation rate was also obtained when PMSG was incorporated into the medium of M199 supplemented with buFF. The rate of maturation was to the tune of 91% when oocytes were matured in buFF alone which was increased non significantly on the addition of PMSG. Highest maturation rate (97%) obtained with M199+buFF (50%v/v)+PMSG did not differ significantly from that obtained by either M199+buFF (10%v/v)+PMSG or buFF+PMSG. It is suggested that buFF alone without any supplementation can form the effective in vitro maturation medium for buffalo oocytes.

Detection of genome-wide structural variations in the Shanghai Holstein cattle population using next-generation sequencing

Dengying Liu,Zhenliang Chen,Zhe Zhang,Hao Sun,Peipei Ma,Kai Zhu,Guanglei Liu,Qishan Wang,Yuchun Pan 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.3

Objective: The Shanghai Holstein cattle breed is susceptible to severe mastitis and other diseases due to the hot weather and long-term humidity in Shanghai, which is the main distribution centre for providing Holstein semen to various farms throughout China. Our objective was to determine the genetic mechanisms influencing economically important traits, especially diseases that have huge impact on the yield and quality of milk as well as reproduction. Methods: In our study, we detected the structural variations of 1,092 Shanghai Holstein cows by using next-generation sequencing. We used the DELLY software to identify deletions and insertions, cn.MOPS to identify copy-number variants (CNVs). Furthermore, we annotated these structural variations using different bioinformatics tools, such as gene ontology, cattle quantitative trait locus (QTL) database and ingenuity pathway analysis (IPA). Results: The average number of high-quality reads was 3,046,279. After filtering, a total of 16,831 deletions, 12,735 insertions and 490 CNVs were identified. The annotation results showed that these mapped genes were significantly enriched for specific biological functions, such as disease and reproduction. In addition, the enrichment results based on the cattle QTL database showed that the number of variants related to milk and reproduction was higher than the number of variants related to other traits. IPA core analysis found that the structural variations were related to reproduction, lipid metabolism, and inflammation. According to the functional analysis, structural variations were important factors affecting the variation of different traits in Shanghai Holstein cattle. Our results provide meaningful information about structural variations, which may be useful in future assessments of the associations between variations and important phenotypes in Shanghai Holstein cattle. Conclusion: Structural variations identified in this study were extremely different from those of previous studies. Many structural variations were found to be associated with mastitis and reproductive system diseases; these results are in accordance with the characteristics of the environment that Shanghai Holstein cattle experience.

Transcriptional regulation of chicken leukocyte cell-derived chemotaxin 2 in response to toll-like receptor 3 stimulation

이석현,이라함,김성조,이학교,나종삼,송기덕 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.12

Objective: Leukocyte cell-derived chemotaxin 2 (LECT2) is associated with several physiological processes including inflammation, tumorigenesis, and natural killer T cell generation. Chicken LECT2 (chLECT2) gene was originally identified as one of the differentially expressed genes in chicken kidney tissue, where the chickens were fed with different calcium doses. In this study, the molecular characteristics and gene expression of chLECT2 were analyzed under the stimulation of toll-like receptor 3 (TLR3) ligand to understand the involvement of chLECT2 expression in chicken metabolic disorders. Methods: Amino acid sequence of LECT2 proteins from various species including fowl, fish, and mammal were retrieved from the Ensembl database and subjected to Insilco analyses. In addition, the time- and dose-dependent expression of chLECT2 was examined in DF-1 cells which were stimulated with polyinosinic:polycytidylic acid (poly [I:C]), a TLR3 ligand. Further, to explore the transcription factors required for the transcription of chLECT2, DF-1 cells were treated with poly (I:C) in the presence or absence of the nuclear factor κB (NFκB) and activated protein 1 (AP-1) inhibitors. Results: The amino acid sequence prediction of chLECT2 protein revealed that along with duck LECT2 (duLECT2), it has unique signal peptide different from other vertebrate orthologs, and only chLECT2 and duLECT2 have an additional 157 and 161 amino acids on their carboxyl terminus, respectively. Phylogenetic analysis suggested that chLECT2 is evolved from a common ancestor along with the actinopterygii hence, more closely related than to the mammals. Our quantitative polymerase chain reaction results showed that, the expression of chLECT2 was up-regulated significantly in DF-1 cells under the stimulation of poly (I:C) (p<0.05). However, in the presence of NFκB or AP-1 inhibitors, the expression of chLECT2 is suppressed suggesting that both NFκB and AP-1 transcription factors are required for the induction of chLECT2 expression. Conclusion: The present results suggest that chLECT2 gene might be a target gene of TLR3 signaling. For the future, the expression pattern or molecular mechanism of chLECT2 under stimulation of other innate immune receptors shall be studied. The protein function of chLECT2 will be more clearly understood if further investigation about the mechanism of LECT2 in TLR pathways is conducted.

Effects of Flaxseed Diets on Fattening Response of Hanwoo Cattle : 2. Fatty Acid Composition of Serum and Adipose Tissues

C. M. Kim,J. H. Kim,T. Y. Chung,박근규 아세아·태평양축산학회 2004 Animal Bioscience Vol.17 No.9

Two separate trials were designed to determine effects of dietary level of whole flaxseed (WFS) on fatty acid composition of serum, and subcutaneous, perirenal, intermuscular, and intramuscular adipose tissues of Korean Hanwoo cattle. Twenty-one bulls (trial 1) and 15 cows (trial 2) were assigned to diets containing 0, 10 or 15% WFS. Relative treatment effects were similar between bulls and cows. The proportion of C18:3 in serum and to a lesser extent in adipose tissues were increased by dietary inclusion of WFS, reflecting supplemented lipid composition of WFS that escaped ruminal biohydrogenation. Animals fed WFS had a lower proportion of saturated fatty acids in serum and adipose tissues than animals fed diets without WFS, while the opposite trend was observed in unsaturated fatty acids with little differences between two WFS groups. WFS-fed animals had higher proportions of C18:1, 18:2, 18:3, 20:3, and 22:3 and lower proportions of C12:0, 14:0, 16:0 and 18:0 in intramuscular fat than animals fed diets without WFS. Furthermore, feeding WFS increased proportions of both w-3 and w-6 fatty acids but decreased the ratio of w-6/w-3 substantially. In conclusion, feeding WFS can be an effective method of increasing absorption of unsaturated fatty acids, and subsequent deposition in adipose tissues.

Comparative evaluation of supplemental zilpaterol hydrochloride sources on growth performance, dietary energetics and carcass characteristics of finishing lambs

A. Rivera-Villegas,A. Estrada-Angulo,B.I. Castro-Perez,J.D. Urias-Estrada,F.G. Rios-Rincon,D. Rodriguez-Cordero,A. Barreras,A. Plascencia,V.M.Gonzalez-Vizcarra,J.F. Sosa-Gordillo,R.A. Zinn 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.2

Objective: We compare the effects of three different approved sources of supplemental zilpaterol on growth-performance responses and carcass characteristics of finishing lambs. Methods: Twenty four Pelibuey×Katahdin lambs (46.75±2.43 kg) were used in a 33-day feeding trial. Lambs were fed a dry rolled corn-based finishing diet. Treatments consisted of the non-supplemental basal diet (Control) versus the basal diet supplemented with 125 mg zilpaterol/kg of diet (as fed basis) from three commercial sources marketed in Mexico: Zilmax (ZIL), Grofactor, and Zipamix. Results: Compared to controls, zilpaterol (ZH) supplementation did not affect dry matter intake (DMI), but increased carcass adjusted daily weight gain (ADG, 36.7%), gain efficiency (34.2%), and dietary net energy (26.0%), and decreased (23.4%) the ratio of observed:expected DMI. Compared to controls, supplemental ZH increased hot carcass weight (6.4%), dressing percentage (3.2%), m. longissimus thoracis (LM) area (15.6%), and shoulder muscle:fat ratio (28.7%), but decreased kidney-pelvic-heart fat, and fat thickness. Supplemental ZH increased 10.9% and 14.3% whole cut weight of loin and leg, respectively, and the proportion (as percentage of cold carcass weight) of leg (4.3%). These increases were reflected in greater forequarter and hindquarter weights. Lambs fed ZH increased (4.6%) empty body weight (EBW) and reduced (14.7%) liver/spleen weight (as g/kg EBW). Likewise, ZH supplementation tended (p = 0.08) to lower (8.9%) visceral fat. Growth performance, energetic efficiency, hot carcass weight, dressing percentage, LM area and whole cuts were not different across supplemental ZH sources. However, compared with non-supplemented controls, only ZIL appreciably decreased carcass fat distribution, including fat thickness, percentage kidney pelvic and heart fat, shoulder fat, and visceral fat. Conclusion: Supplemental ZH increases ADG, gain efficiency, carcass dressing percentage, and LM area. The magnitude of these responses was similar among ZH sources. Nevertheless, compared with non-supplemented controls, only ZIL appreciably decreases carcass fat. The basis for this is uncertain, but indicative that some practical differences in zilpaterol bio-equivalency may exist across commercial sources tested.

Genetic evaluation of eggshell color based on additive and dominance models in laying hens

Guo Jun,Wang Kehua,Qu Liang,Dou Taocun,Ma Meng,Shen Manman,Hu Yuping 아세아·태평양축산학회 2020 Animal Bioscience Vol.33 No.8

Objective: Eggshells with a uniform color and intensity are important for egg production because many consumers assess the quality of an egg according to the shell color. In the present study, we evaluated the influence of dominant effects on the variations in eggshell color after 32 weeks in a crossbred population. Methods: This study was conducted using 7,878 eggshell records from 2,626 hens. Heritability was estimated using a univariate animal model, which included inbreeding coefficients as a fixed effect and animal additive genetic, dominant genetic, and residuals as random effects. Genetic correlations were obtained using a bivariate animal model. The optimal diagnostic criteria identified in this study were: L* value (lightness) using a dominance model, and a* (redness), and b* (yellowness) value using an additive model. Results: The estimated heritabilities were 0.65 for shell lightness, 0.42 for redness, and 0.60 for yellowness. The dominance heritability was 0.23 for lightness. The estimated genetic correlations were 0.61 between lightness and redness, –0.84 between lightness and yellowness, and –0.39 between redness and yellowness. Conclusion: These results indicate that dominant genetic effects could help to explain the phenotypic variance in eggshell color, especially based on data from blue-shelled chickens. Considering the dominant genetic variation identified for shell color, this variation should be employed to produce blue eggs for commercial purposes using a planned mating system.

Genetic architecture and candidate genes detected for chicken internal organ weight with a 600 K single nucleotide polymorphism array

Taocun Dou,Manman Shen,Meng Ma,Liang Qu,Yongfeng Li,Yuping Hu,Jian Lu,Jun Guo,Xingguo Wang,Kehua Wang 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.3

Objective: Internal organs indirectly affect economic performance and well-being of animals. Study of internal organs during later layer period will allow full utilization of layer hens. Hence, we conducted a genome-wide association study (GWAS) to identify potential quantitative trait loci or genes that potentially contribute to internal organ weight. Methods: A total of 1,512 chickens originating from White Leghorn and Dongxiang Blue-Shelled chickens were genotyped using high-density Affymetrix 600 K single nucleotide polymorphism (SNP) array. We conducted a GWAS, linkage disequilibrium analysis, and heritability estimated based on SNP information by using GEMMA, Haploview and GCTA software. Results: Our results displayed that internal organ weights show moderate to high (0.283 to 0.640) heritability. Variance partitioned across chromosomes and chromosome lengths had a linear relationship for liver weight and gizzard weight (R2 = 0.493, 0.753). A total of 23 highly significant SNPs that associated with all internal organ weights were mainly located on Gallus gallus autosome (GGA) 1 and GGA4. Six SNPs on GGA2 affected heart weight. After the final analysis, five top SNPs were in or near genes 5-Hydroxytryptamine receptor 2A, general transcription factor IIF polypeptide 2, WD repeat and FYVE domain containing 2, non-SMC condensin I complex subunit G, and sonic hedgehog, which were considered as candidate genes having a pervasive role in internal organ weights. Conclusion: Our findings provide an understanding of the underlying genetic architecture of internal organs and are beneficial in the selection of chickens.

Carcass characteristics of lambs fed diets with increasing levels of crude glycerin

Caio Alves da Costa,Francisco Fernando Ramos de Carvalho,Adriana Guim,Gilcifran Prestes de Andrade,Daniel Barros Cardoso,Michel do Vale Maciel,Gabriela Goncalves da Silva,Andreza Guedes de Oliveira Na 아세아·태평양축산학회 2019 Animal Bioscience Vol.32 No.12

Objective: An experiment was conducted to evaluate the effects of increasing levels of crude glycerin (0%, 6%, 12%, and 18%) used as a substitute for corn in lamb feed on the quantitative characteristics of the carcass. Methods: A total of 40 crossbred Santa Inês lambs that were four months old with a mean initial weight of 21.0±0.8 kg were randomly distributed in four treatments with ten replicates. The animals were slaughtered after 66 days of confinement. The effects of crude glycerin as a replacement for corn in the diet of the lambs on the carcass characteristics, commercial cut weight and yield and carcass measurements were studied. Results: There was an increasing linear effect for body weight at slaughter with the replacement of corn by crude glycerin. The dry matter and metabolizable energy intakes, weight of the empty body, hot carcass weight and cold carcass weight showed a quadratic effect, with maximum crude glycerin levels estimated at 10.9%, 9.8%, 10.83%, 11.78%, and 11.35%, respectively. The initial pH was not influenced by the replacement of corn for crude glycerin, while the final pH presented a quadratic effect. The other parameters of the carcass and the weights and yields of commercial cuts were not influenced. There was also no effect of the diets on carcass morphometric measurements, except for the thoracic perimeter and the carcass compactness index, which presented quadratic and linear effects, respectively. Conclusion: Crude glycerin can replace up to 18% of corn because it favours muscle tissue deposition without promoting changes in the main carcass characteristics of lambs.