EDG1 Genes Research Articles

Effect of Expressions and SNPs of Candidate Genes on Intramuscular Fat Content in Qinchuan Cattle.

Simple SummaryIntramuscular fat (IMF), which characterizes the marbling, is a key meat quality trait in the beef industry. In this study, we validated the effect of the expression levels and single nucleotide polymorphisms (SNPs) of AKIRIN2, TTN, EDG1, and MYBPC1 genes on IMF content in Chinese Qinchuan cattle. The results showed that the expression levels of these genes could affect the IMF content, and the SNP in EGD1 could affect the IMF content in Qinchuan. The information of this study may be applied to effective marker-assisted selection, to increase the levels of marbling in Qinchuan beef production.Marbling is characterized by the amount and distribution of intramuscular fat (IMF). The AKIRIN2, TTN, EDG1, and MYBPC1 genes are well-known marbling-related genes, which were first identified in Japanese Black beef cattle. The objectives of this study were to analyze the correlation of the expression levels of these genes in the longissimus muscle (LM) with IMF content, and the associations between the single nucleotide polymorphisms (SNPs) in these genes and IMF content in Chinese Qinchuan cattle (n = 350). The association analyses showed that the g.42041062G>T SNP in the EDG1 gene was significantly associated with IMF content in Qinchuan (p < 0.05). Further, the expressions of the EDG1 and MYBPC1 were up-regulated (p < 0.05) in LM of Qinchuan cattle group with low IMF content. Down-regulations of the AKIRIN2 and TTN genes (p < 0.05 and p < 0.01, respectively) were observed in the Qinchuan cattle group with high IMF content. These results suggest possible effects of the expression levels of selected genes on IMF content in the LM, and the g.42041062G>T SNP in the EDG1 gene might be useful as a molecular marker for IMF content in Qinchuan.

The The favorable alleles of AKIRIN2:c.*188G&gt;A, EDG1:c.-312A&gt;G and TTN:g.231054C&gt;T as candidate markers for high-marbling are very low in Bali cattle

Abstract. Anwar S, Wulandari AS, Putra WPB, Said S. 2019. The favorable alleles of AKIRIN2:c.*188G&gt;A, EDG1:c.-312A&gt;G and TTN:g.231054C&gt;T as candidate markers for high-marbling are very low in Bali cattle. Biodiversitas 20: 965-970. Marbling is one of primary factors influencing the palatability of beef. The AKIRIN2, EDG1 and TTN genes are considered as the three candidate genes responsible for high-marbling in beef cattle. This study aimed to investigate the genetic polymorphism of the SNPs of AKIRIN2:c.*188G&gt;A, EDG1:c.-312A&gt;G and TTN g.231054C&gt;T in Bali cattle. A total of 125 DNA samples of Bali cattle were genotyped for AKIRIN2 and TTN gene and 82 DNA samples for EDG1 gene using PCR-RFLP method. In the present study, it was found that the A allele of AKIRIN2:c.*188G&gt;A, the G allele of EDG1:c.-312A&gt;G and the T allele of TTN g.231054C&gt;T that considered as the favorable alleles were found to be very low in Bali cattle (0.004, 0.000, and 0.004, respectively). This results indicated that the polymorphism of AKIRIN2:c.*188G&gt;A, EDG1:c.-312A&gt;G and TTN g.231054C&gt;T were very low and can be considered as monomorphic in Bali cattle. These findings suggested that Bali cattle may not genetically potential as a high-marbling cattle breed. However, the other specific genes for high-marbling still needs to be investigated in Bali cattle.

Polymorphism of ADIPOQ and EDG1 genes in Indonesian beef cattle

The ADIPOQ and EDG1 genes were responsible in intramuscular fat deposition and marbling scores. This study was aimed to identify polymorphism of indel g.81966364D&gt;I in promoter region of ADIPOQ gene and SNP c.-312A&gt;G in 5' UTR of EDG1 gene in Indonesian beef cattle. Blood samples were collected from 211 cattle, including Bali (44), Madura (20), Pesisir (18), Katingan (20), PO (22), Pasundan (20), SO (12), Brahman (20), Simmental (15) and Limousin (18). Polymorphism of ADIPOQ gene was analyzed using PCR and direct sequencingmethods, whereas EDG1 gene was analyzed using PCR-RFLP (MscI enzyme) and direct sequencingmethods. Results of genotyping indel g.81966364D&gt;I was monomorphic (DD genotype). The SNP c.-312A&gt;G was polymorphic (AA and AG genotype) in Madura, Pesisir, Pasundan, Brahman, and Limousine. The Frequencies of allele A and G were 0.95, 0.92, 0.98, 0.95, 0.94 and 0.05, 0.08, 0.02, 0.05, 0.06 respectively. The values of Ho and He were 0.05-0.17 and 0.05-0.15 respectively and in Hardy-Weinberg equilibrium (P&gt;0.05). In Bali, Katingan, PO, SO and Simmental were monomorphic (GG genotype). In Bali cattle, two novel SNP candidates were found in position of c.-399C&gt;T and c.-273C&gt;G which were potential to be used as genetic markers of marbling score for Bali cattle. As result this study, it can be concluded that ADIPOQ gene was similar while EDG1 gene was different in Indonesian beef cattle. in addition, found two candidates potential SNP in Bali cattle.

Genetic characterization of Vietnamese Yellow cattle using mitochondrial DNA and Y-chromosomal haplotypes and genes associated with economical traits.

Vietnamese Yellow cattle are native cattle well adapted to local tropical environment. The aim of this study was to investigate genetic characteristics of the Yellow cattle using molecular markers. We investigated the nucleotide sequences of mitochondrial DNA and SRY gene on Y chromosome, and genotyped SREBP-1, SCD1, EDG1, NCAPG, DGAT1, MC1R, and HSP70 genes in the Yellow cattle population. The sequence analysis of the mitochondrial DNA showed that most of the cattle possesses zebu (Bos indicus) type I1 haplotype, suggesting relatively low genetic diversity in maternal lineage. The sequence analysis of the SRY gene indicates that while most of the males possess zebu type haplotype, taurine (Bos taurus) type haplotype was also observed, suggesting gene-flow from taurine cattle. The results of the genotyping of the functional genes showed that the NCAPG, SCD, MC1R, and HsSP70 genes are polymorphic in the population, whereas the SREBP-1, EDG1, and DGAT1 genes are monomorphic. Particularly, the presence of the desirable and undesirable alleles of the NCAPG and HSP70 genes, respectively, will be important for the selection of animals by potential performances in meat productivity and fertility. The present findings will be informative for future conservation and breeding of the Vietnamese Yellow cattle.

Molecular genetics and bioinformatics analysis of EDG1 and AKIRIN2 genes in Iranian fat-tailed and nonfat-tailed sheep breeds

This study was conducted to detect different allelic forms and its association with fat-tail traits, and also investigation of domains variants in 5/- and 3/-UTR regions of EDG1 and AKIRIN2 genes in Iranian fat-tailed (Lori-Bakhtiari) and nonfat-tailed (Zel) sheep breeds. Blood samples were randomly collected from 300 sheep (150 samples per each breed) and genomic DNA was extracted by modified salting out method. Two primer pairs were designed by oligo7 software for amplification of fragments with the length of 516 and 454bp from EDG1 and AKIRIN2 genes, respectively. The results of genotyping by SSCP assay and silver nitrate staining showed that, in both breeds, there was no incidence of mutation in AKIRIN2 gene. But, the EDG1 was highly polymorph in studied sheep breeds, so that, six banding patterns with different frequencies of A (36.73 and 31.19%), B (8.84 and 21.6%), C (7.48 and 41.12%), D (29.93 and 34.50%), E (13.61 and 34.10%) and F (3.40 and 38.1%) were observed for Zel and Lori-Bakhtiari breeds, respectively. Comparison of banding pattern frequencies by Chi-square test showed significant difference between two breeds (P<0.01). One sample from each banding patterns were sequenced. The alignment and motif analysis of sequences by BioEdit and DNASIS MAX software showed incidence of 11 single nucleotide polymorphism (SNP) and 18 DNA motifs in EDG1 marker site. According to different SNPs, the paternal and maternal haplotypes were determined by phase software. The association study showed significant association between EDG1 gene haplotypes and banding patterns with fat-tail traits in Lori-Bakhtiari sheep except for height of fat-tail. This study was the first effort for evaluate of these genes in sheep. Therefore, more studies on other breeds of sheep and on coding sequence of these genes are recommended.

Effect of Single Nucleotide Polymorphism of Endothelial Differentiation G-Protein Coupled Receptor 1 (EDG1) Gene on Marbling Score in Hanwoo

Marbling (intramuscular fat) is the most economically important meat quality trait in Hanwoo (Korean cattle). The endothelial differentiation G-protein coupled receptor 1 (EDG1) gene, involved in blood vessel formation, is located within the genomic region of a quantitative trait locus (QTL) for marbling on bovine chromosome 3. Thus, the EDG1 gene can be considered as a positional and functional candidate gene for meat quality in beef cattle. This study aimed to identify single nucleotide polymorphisms (SNPs) in the EDG1 gene and to evaluate their associations with carcass traits in Hanwoo population. We have sequenced a fragment of 5'-UTR of the EDG1 gene and identified one SNP. Genotyping of the g.166A>G SNP marker was carried out using PCR-RFLP analysis in 309 Hanwoo steers in order to evaluate their association with carcass traits. The g.166A>G SNP marker showed a significant effect on the marbling score. Animals with the GG genotype had higher marbling score compared with AA and AG genotypes (p<0.05). This SNP marker also showed a significant additive effects for the marbling score (p<0.05). These results suggest that the EDG1 gene can be used as a molecular marker for DNA marker-assisted selection in order to increase the levels of the marbling score in Hanwoo.

한우에서 TG와 EDG1 유전자의 단일염기다형 확인 및 도체형질과의 연관성 분석

Thyroglobulin (TG) gene was known to be regulated fat cell growth and differentiation and the endothelial differentiation sphingolipid G-protein-coupled receptor 1 (EDG1) gene involves blood vessel formation and known to be affecting carcass traits in beef cattle. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) in both TG and EDG1 genes and to analyze the association with carcass traits in Korean cattle (Hanwoo). The T354C SNP in TG gene located at the 3' flanking region and c.-312A>G SNP located at 3'-UTR of EDG1 gene were used for genotyping the animals using PCR-RFLP method. Three genotypes were identified in T354C SNP in TG gene and only two AA and AG genotypes were observed for the c.-312A>G SNP in EDG1 gene. The results indicated that T354C SNP in TG gene was not significantly associated with carcass traits. However, the c.-312A>G SNP in EDG1 gene had significant effects on backfat thickness (BF) and yield index (YI). These results may provide valuable information for further candidate gene studies affecting carcass traits in Korean cattle and may use as marker assisted selection for improving the quality of meat in Hanwoo.

Abstract P261: Endothelial Differentiation Gene-1 Is Involved in Related Transcriptional Enhancer Factor-1--Induced Endothelial Cell Aggregation

We have previously demonstrated that Related Transcriptional Enhancer Factor-1 (RTEF-1) regulated angiogenesis through induction of vascular endothelial growth factor (VEGF) under hypoxic conditions. Recently, we discovered that RTEF-1 accelerated endothelial cell connections and enhanced endothelial cell aggregation. To investigate the mechanism of RTEF-1 in regulated endothelial aggregation, a DNA array analysis was performed in the endothelial cell (HMEC-1) lacking RTEF-1 by its siRNA and resulted in a significant decrease of endothelial differentiation gene -1 (Edg-1). RTEF-1 significantly increase Edg-1 promoter activity (4.10 Fold) and the expression levels of Edg-1 gene are concordant with the RTEF-1 gene in both mRNA and protein levels in cultured endothelial cells as well as in endothelial specific RTEF-1 transgenic mice (VE-Cad-RTEF-1) and RTEF-1 knock out mice (Tie2 Cre :RTEF-1 floxp ) . In Matrigel tube formation assay, over-expression of Edg-1 gene stimulated endothelial cell aggregation, while siRNA of Edg-1 significantly blocked RTEF-1-driven endothelial cell aggregation. Furthermore, using an Edg-1 receptor agonist Sphingosine-1-phosphate (S1P), the acceleration of the proliferation, migration and tube formation were observed, whereas Edg-1 receptor inhibitor Pertussis Toxin (PTX) could block RTEF-1-induced cell aggregation. Our data demonstrates that Edg-1 is a potential target gene of RTEF-1. It is involved in RTEF-1 regulated endothelial cell angiogenesis, specifically in endothelial cell aggregation.

Recovery and Expression of Messenger RNA from Postmortem Human Brain Tissue

The Bryan Alzheimer's Disease Research Center Rapid Autopsy Program at Duke University Medical Center obtains postmortem human brain tissue for experimental investigations. We evaluated 19 brains for RNA integrity and mRNA gene expression. Nine were from patients diagnosed with Alzheimer's disease, and ten were from nondemented controls. In all cases, the following variables were recorded: postmortem procurement delay (range, 1 hour and 10 minutes to 14 hours), pH of cerebrospinal fluid, premortem fever or sepsis, provision of supplemental oxygen in the agonal period, and temporal relation to time of death (either sudden death or protracted illness). Total RNA was extracted, quantified, and evaluated by agarose gel electrophoresis and quantitative gene expression analysis of 18S rRNA and edg-1 using TaqMan technology. All samples appeared to yield intact RNA without significant degradation, and expression of the edg-1 gene was detected by the real time reverse transcriptase polymerase chain reaction in all cases. We conclude that intact RNA can be obtained from postmortem human brain tissue, even in patients with severe premortem illnesses and delayed postmortem tissue procurement intervals. However, we caution that the successful expression of certain genes from postmortem brain tissue may require enhanced procurement efforts to maximize RNA integrity.

Sphingosine 1-phosphate receptors

Sphingosine-1-phosphate (SPP) is a bioactive lipid produced from the metabolism of sphingomyelin. It is an important constituent of serum and regulates cell growth, survival, migration, differentiation and gene expression. Its mode of action has been enigmatic; however, recent findings have shown that a family of G-protein-coupled receptors (GPCR) of the endothelial differentiation gene (EDG) family serve as plasma membrane-localized receptors for SPP. Furthermore, the EDG receptors appear to be SPP receptor subtypes with distinct signaling characteristics. In vascular endothelial cells, SPP acts on EDG-1 and EDG-3 subtypes of receptors to induce cell survival and morphogenesis. Such pathways appear to be critical for SPP-induced angiogenic response in vivo. In addition, the EDG-1 gene is essential for vascular maturation in development. Moreover, developmental studies in Zebrafish have indicated that SPP signaling via the EDG-5 like receptor Miles Apart ( Mil) is essential for heart development. These data strongly suggest that a physiological role of SPP is in the formation of the cardiovascular system. Despite these recent findings, much needs to be clarified with respect to the physiological role of SPP synthesis and action. This review will focus on the recent findings on SPP receptors and the effects on the cardiovascular system.

Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation.

Sphingolipid signaling pathways have been implicated in many critical cellular events. Sphingosine-1-phosphate (SPP), a sphingolipid metabolite found in high concentrations in platelets and blood, stimulates members of the endothelial differentiation gene (Edg) family of G protein-coupled receptors and triggers diverse effects, including cell growth, survival, migration, and morphogenesis. To determine the in vivo functions of the SPP/Edg signaling pathway, we disrupted the Edg1 gene in mice. Edg1(-/-) mice exhibited embryonic hemorrhage leading to intrauterine death between E12.5 and E14.5. Vasculogenesis and angiogenesis appeared normal in the mutant embryos. However, vascular maturation was incomplete due to a deficiency of vascular smooth muscle cells/pericytes. We also show that Edg-1 mediates an SPP-induced migration response that is defective in mutant cells due to an inability to activate the small GTPase, Rac. Our data reveal Edg-1 to be the first G protein-coupled receptor required for blood vessel formation and show that sphingolipid signaling is essential during mammalian development.

The Inducible G Protein-coupled Receptor edg-1 Signals via the Gi/Mitogen-activated Protein Kinase Pathway

The edg-1 gene encodes an inducible G protein-coupled receptor (GPR) homologue that is induced during the in vitro differentiation of human endothelial cells. The aim of this study was to investigate the G protein-coupling and -signaling properties of the edg-1 polypeptide. The third cytosolic loop (i3) of edg-1 associates with G(i) alpha and G(o) alpha polypeptides in a guanosine 5'-O-(thiotriphosphate)-sensitive manner. Immunoprecipitation of the edg-1 polypeptide in transfected cells results in the co-precipitation of G(i) alpha 1 and G(i) alpha 3 polypeptides. These data strongly suggest that edg-1 is capable of coupling to the Gi pathway. Overexpression of the edg-1 GPR in human embryonic kidney 293 cells results in the sustained activation of the MAP kinase activity that is blocked by pertussis toxin treatment. Moreover, NIH3T3 cells permanently transfected with edg-1 exhibit enhanced MAP kinase and phospholipase A2 activities. These data suggest that the G(i)/mitogen-activated protein kinase pathway is a major signaling pathway regulated by the orphan receptor edg-1.

Cloning of the rat edg-1 immediate-early gene: expression pattern suggests diverse functions

The edg-1 immediate-early gene encodes a G-protein-coupled receptor homolog implicated in endothelial cell differentiation. We report the cloning of the rat edg-1 gene. Our Northern analyses indicate that edg-1 is much more widely expressed than previously thought, edg-1 mRNA was found in many organs at several stages of development with relatively high levels present in adult brain, edg-1 transcripts were also detected in several cell lines. Expression of edg-1 mRNA in the PC 12 cell model of neuronal differentiation was unaffected by agents that cause PC 12 cells to differentiate or proliferate. Therefore, edg-1 may play a cell-type-specific role in differentiation and also participate in neuro-transmission.