A Novel Discipline in Embryology — Animal Embryo Breeding

The modern animal biotechnologies, such as animal cloning, transgenesis, sex determination, stem cells, designing new livestock, must be performed on animal gametes including sperm and oocytes, and embryos based on embryology theory. Currently, some key biotechnologies in embryology have become the most powerful tool for animal scientists and breeders to improve genetic construction of animal herds. Here, authors put forward a new concept of Animal Embryo Breeding Science to describe this discipline formation, development, and application in animal genetic improvement and breeding. The relationship of embryo breeding with other disciplines has been profiled. Thus, animal scientists and breeders can easily understand and apply embryo breeding theory and related key techniques to accelerate animal improvement speed, to modify genetic construction of animal population, and to design and create new animal individual or breed

Identification of SNPs in chemerin gene and association with carcass and meat quality traits of Qinchuan Cattle

Chemerin is a novel adipokine that regulates adipogenesis and adipocyte metabolism via its own receptor. In this study, two novel SNPs (868A>G in exon 2 and 2692C>T in exon 5) of chemerin gene were identified by PCR-SSCP and DNA sequencing technology. The allele frequencies of the novel SNPs were determined in the genetically diverse bovine breeds including six Chinese indigenous cattle breeds (Caoyuan red, Jiaxian red, Luxi, Nanyang, Qinchuan and Xia’nan cattle). We evaluated the potential association of the SNPs with traits measured by ultrasound measurement in 214 Qinchuan individuals. Furthermore, meat quality traits data gotten from carcass measurement in another 69 Qinchuan individuals were analyzed by the comparison between the genotypes and their phenotypic data. Results showed that SNP 868A>G had a significant association with the ultrasound loin-muscle area (P < 0.05), loin-eye area and water holding capability (P < 0.05). And also revealed significant effects of genotype on the ultrasound backfat thickness (P < 0.05), backfat thickness and water holding capability (P < 0.05) of SNP 2692C>T. It was shown that associations do exist between chemerin gene and carcass and meat quality traits. As a result of the small sample size of this study, it is proposed that further effort is required to validate these findings in larger populations. It could be concluded that ultrasound measurements are similar in accuracy to carcass measurements for predicting carcass and meat quality traits in cattle, and could be a useful predictor of retail yield in live animals.Keywords: Bos bovine, chemerin gene, PCR-SSCP, SNP, meat quality trait

Molecular characterization, polymorphism of growth differentiation factor 5 gene and association with ultrasound measurement traits in native Chinese cattle breeds

Growth differentiation factor 5 (GDF5), involved in the development and maintenance of bone andcartilage, is a n important candidate gene for growth and carcass traits selection through markerassisted selection (MAS). Genomic structural analysis showed that bovine GDF5 shares much similarity with human GDF5. The latest findings demonstrate that the single nucleotide polymorphism (SNP) T586C in exon 1 is significantly associated with ultrasound marbling score (UMAR) and ultrasound backfat thickness (UBF). Furthermore, the analysis of T586C SNP marker shows there are significant effects on the UBF (P = 0.0498) and on the UMAR (P = 0.0058) in 465 individuals. These results clearly suggest that the GDF5 gene is among target genes for carcass traits in bovine reproduction and breeding.Keywords: Cattle, GDF5 gene, ultrasound measurement, polymorphism, association analysisAfrican Journal of Biotechnology Vol. 9(33), pp. 5269-5273, 16 August, 201

OLFML3 Expression is Decreased during Prenatal Muscle Development and Regulated by MicroRNA-155 in Pigs

The Olfactomedin-like 3 (OLFML3) gene has matrix-related function involved in embryonic development. MicroRNA-155 (miR-155), 21- to 23-nucleotides (nt) noncoding RNA, regulated myogenesis by target mRNA. Our LongSAGE analysis suggested that OLFML3 gene was differently expressed during muscle development in pig. In this study, we cloned the porcine OLFML3 gene and detected its tissues distribution in adult Tongcheng pigs and dynamical expression in developmental skeletal muscle (12 prenatal and 10 postnatal stages) from Landrace (lean-type) and Tongcheng (obese-type) pigs. Subsequently, we analyzed the interaction between OLFML3 and miR-155. The OLFML3 was abundantly expressed in liver and pancreas, moderately in lung, small intestine and placenta, and weakly in other tissues and postnatal muscle. There were different dynamical expression patterns between Landrace and Tongcheng pigs during prenatal skeletal muscle development. The OLFML3 was down-regulated (33-50 days post coitus, dpc), subsequently up-regulated (50-70 dpc), and then down-regulated (70-100 dpc) in Landrace pigs, while in Tongcheng pigs, it was down-regulated (33-50 dpc), subsequently up-regulated (50-55 dpc) and then down-regulated (55-100 dpc). There was higher expression in Tongcheng than Landrace in prenatal muscle from 33 to 60 dpc, and opposite situation from 65 to 100 dpc. Dual luciferase assay and real time PCR documented that OLFML3 expression was regulated by miR-155 at mRNA level. Our research indicated that OLFML3 gene may affect prenatal skeletal muscle development and was regulated by miR-155. These finding will help understanding biological function and expression regulation of OLFML3 gene in mammal animals

Estimation of genetic parameters for fertility traits in Chinese Holstein of south China

Introduction: Chinese Holstein in South China suffer heat stress for a long period, which leads to evolutionary differences from Chinese Holstein in North China. The aim of this study was to estimate the genetic parameters of fertility traits for Chinese Holstein in South China.Methods: A total of 167,840 Chinese Holstein heifers and cows from Guangming Animal Husbandry Co., LTD farms were used in this study. The fertility traits analyzed were calving interval (CI), days open (DO), age of first service (AFS), age of first calving (AFC), calving to first insemination (CTFS), first insemination to conception (FSTC), gestation length (GL), non-return rate to 56 days (NRR), and number of services (NS).Results: The descriptive statistics revealed that the same trait in heifers performed better than in cows, which was consistent with the other studies. The heritabilities of fertility traits in this study ranged from close to 0 (for NS of cows) to 0.2474 (for AFC of heifers). The genetic correlation of NRR between heifers and cows was 0.9993, which indicates that the NRR for heifers and cows could be treated as one trait in this population.Conclusion: The heritabilities of fertility traits in Chinese Holstein in south China were quite different from the heritabilities of fertility traits in North China. NRR56, NS, AFC, and CI were suggested to be included into the selection index to improve fertility performance of Chinsese Holstein of south China. The results of this study could provide genetic parameters for the animal breeding program of Chinese Holstein in the south of China

Polymorphisms of the bovine chemokine receptor-like 1 gene and their associations with meat quality traits in Qinchuan cattle ( Bos taurus )

Background: We investigated the polymorphisms of the bovine chemokine receptor-like 1(CMKLR1) gene. The coding region of CMKLR1 was screened in Qinchuan cattle by PCR-RFLP technology. Results: In this study, we discovered two single nucleotide polymorphisms (SNPs) (264G > C and 762C > T) in the coding region of the CMKLR1 gene. Hence, we described the BmgT120l and Pdm1 PCR-RFLP methods for detecting the 64G > C and 762C > T mutations, respectively. PCR-RFLP and sequencing were used to analyze the two loci of CMKLR1 gene in 324 individuals, which were randomly selected from breeding populations. Furthermore, meat quality traits in another 80 Qinchuan individuals were analyzed by the comparison between the genotypes and their phenotypic data. Conclusions: The results showed that the G264C SNP and C762T SNP of bovine CMKLR1 were significantly associated with backfat thickness (BFT) and water holding capacity (WHC), respectively

Functional Genomic Analysis of Variation on Beef Tenderness Induced by Acute Stress in Angus Cattle

Beef is one of the leading sources of protein, B vitamins, iron, and zinc in human food. Beef palatability is based on three general criteria: tenderness, juiciness, and flavor, of which tenderness is thought to be the most important factor. In this study, we found that beef tenderness, measured by the Warner-Bratzler shear force (WBSF), was dramatically increased by acute stress. Microarray analysis and qPCR identified a variety of genes that were differentially expressed. Pathway analysis showed that these genes were involved in immune response and regulation of metabolism process as activators or repressors. Further analysis identified that these changes may be related with CpG methylation of several genes. Therefore, the results from this study provide an enhanced understanding of the mechanisms that genetic and epigenetic regulations control meat quality and beef tenderness

miRNA-dysregulation associated with tenderness variation induced by acute stress in Angus cattle

miRNAs are a class of small, single-stranded, non-coding RNAs that perform post-transcriptional repression of target genes by binding to 3’ untranslated regions. Research has found that miRNAs involved in the regulation of many metabolic processes. Here we uncovered that the beef quality of Angus cattle sharply diversified after acute stress. By performing miRNA microarray analysis, 13 miRNAs were significantly differentially expressed in stressed group compared to control group. Using a bioinformatics method, 135 protein-coding genes were predicted as the targets of significant differentially expressed miRNAs. Gene Ontology (GO) term and Ingenuity Pathway Analysis (IPA) mined that these target genes involved in some important pathways, which may have impact on meat quality and beef tenderness.https://doi.org/10.1186/2049-1891-3-1

Function and Transcriptional Regulation of Bovine TORC2 Gene in Adipocytes: Roles of C/EBPγ, XBP1, INSM1 and ZNF263

The TORC2 gene is a member of the transducer of the regulated cyclic adenosine monophosphate (cAMP) response element binding protein gene family, which plays a key role in metabolism and adipogenesis. In the present study, we confirmed the role of TORC2 in bovine preadipocyte proliferation through cell cycle staining flow cytometry, cell counting assay, 5-ethynyl-20 -deoxyuridine staining (EdU), and mRNA and protein expression analysis of proliferation-related marker genes. In addition, Oil red O staining analysis, immunofluorescence of adiponectin, mRNA and protein level expression of lipid related marker genes confirmed the role of TORC2 in the regulation of bovine adipocyte differentiation. Furthermore, the transcription start site and sub-cellular localization of the TORC2 gene was identified in bovine adipocytes. To investigate the underlying regulatory mechanism of the bovine TORC2, we cloned a 1990 bp of the 5’ untranslated region (50UTR) promoter region into a luciferase reporter vector and seven vector fragments were constructed through serial deletion of the 50UTR flanking region. The core promoter region of the TORC2 gene was identified at location −314 to −69 bp upstream of the transcription start site. Based on the results of the transcriptional activities of the promoter vector fragments, luciferase activities of mutated fragments and siRNAs interference, four transcription factors (CCAAT/enhancer-binding protein C/BEPγ, X-box binding protein 1 XBP1, Insulinoma-associated 1 INSM1, and Zinc finger protein 263 ZNF263) were identified as the transcriptional regulators of TORC2 gene. These findings were further confirmed through Electrophoretic Mobility Shift Assay (EMSA) within nuclear extracts of bovine adipocytes. Furthermore, we also identified that C/EBPγ, XBP1, INSM1 and ZNF263 regulate TORC2 gene as activators in the promoter region. We can conclude that TORC2 gene is potentially a positive regulator of adipogenesis. These findings will not only provide an insight for the improvement of intramuscular fat in cattle, but will enhance our understanding regarding therapeutic intervention of metabolic syndrome and obesity in public health as well