Expression stability of reference genes in the skeletal muscles of beef cattle

Mberema, Christopher Heinz Hunke; Sparagano, Olivier

doi:10.5897/ajb2016.15140

Cited by 4 publications

(2 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In cattle, De Ketelaere et al (2006) selected sdha , ywhaz , and 18S rRNA as being the most stable genes for the accurate normalization of qPCR of bovine polymorphonuclear leukocytes [ 35 ]. Likewise, sdha has also been ranked greatest in terms of expression stability in bovine neutrophils [ 36 ]. However, the reference genes mentioned previously were described for different experimental conditions.…”

Section: Discussionmentioning

confidence: 99%

Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

Lozano-Villegas

Rodríguez‐Hernández

Herrera‐Sánchez

et al. 2021

Animals

View full text Add to dashboard Cite

Real-time PCR is widely used to study the relative abundance of mRNA due to its specificity, sensitivity, and repeatability quantification. However, relative quantification requires a reference gene, which should be stable in its expression, showing lower variation by experimental conditions or tissues. The aim of this study was to evaluate the stability of the expression of five commonly used reference genes (actb, ywhaz, b2m, sdha, and 18s rRNA) at different physiological stages (alert and emergency) in three different cattle breeds. In this study, five genes (actb, ywhaz, b2m, sdha, and 18s rRNA) were selected as candidate reference genes for expression studies in the whole blood from three cattle breeds (Romosinuano, Gyr, and Brahman) under heat stress conditions. The transcription stability of the candidate reference genes was evaluated using geNorm and NormFinder. The results showed that actb, 18SrRNA, and b2m expression were the most stable reference genes for whole blood of Gyr and Brahman breeds under two states of livestock weather safety (alert and emergency). Meanwhile, actb, b2m, and ywhaz were the most stable reference genes for the Romosinuano breed.

show abstract

Section: Discussionmentioning

confidence: 99%

Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

Lozano-Villegas

Rodríguez‐Hernández

Herrera‐Sánchez

et al. 2021

Animals

View full text Add to dashboard Cite

show abstract

“…Fluorescence was monitored during PCR by BIO‐RAD Real‐Time System (CFX96). Data were analyzed and normalized to GAPDH expression as internal control, which was reported to be stable in cattle muscle ( M values < 1.5; Mberema & Sparagano, 2017). The expressions of target gene were calculated from ∆Cq of sample divide by ∆Cq of calibrator as ∆∆Cq using Bio‐Rad CFX Mastero 1.1 (Version 4.1.2433.12219).…”

Section: Methodsmentioning

confidence: 99%

Effects of body condition at far‐off dry period on blood biochemistry, liver triacylglycerol and muscular monocarboxylate transporter‐1 mRNA expression in tropical Holstein dairy cows during peripartum period

Triwutanon

Rukkwamsuk

2021

Animal Science Journal

View full text Add to dashboard Cite

Effects of body condition score (BCS) at far-off dry period on blood biochemistry, liver triacylglycerol, and muscular monocarboxylate transporter-1 mRNA expression in tropical Holstein dairy cows during peripartum period were studied. Eight dry cows were divided into two groups of four cows based on their BCS at 8 weeks prepartum: high BCS (HBS) and low BCS (LBS). Blood samples were collected weekly from 8 weeks before expected calving date until 8 weeks postpartum for determination of serum glucose, non-esterified fatty acid (NEFA), and β-hydroxybutyrate (BHBA).Muscle and liver samples were biopsied at 2 weeks before expected calving date and at 2 weeks postpartum for determination of muscular monocarboxylate transporter-1 (MCT-1) mRNA expression and liver triacylglycerol concentrations.Decreased BCS and elevated serum NEFA and BHBA indicated that LBS cows entered to the NEB period earlier than did HBS cows. LBS cows showed lower liver TAG and higher muscular MCT-1 mRNA expression than did HBS cows. The MCT-1 mRNA expression at 2 weeks prepartum was negatively correlated with BCSs at 2-3 weeks prepartum and at 1-2 weeks postpartum. In conclusion, LBS cows most likely adapted to NEB by upregulation of muscular MCT-1 expression and had lower NEB consequences during postpartum period.

show abstract

Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle

Coelho¹,

Chalfun-Júnior²,

Barreto³

et al. 2022

Revista Brasileira De Zootecnia

View full text Add to dashboard Cite

The objective of the present study was to evaluate the stability of candidate reference genes and select the genes that can be used for normalizing realtime polymerase chain reaction (PCR) in the liver, skeletal muscle, and jejunum tissues of Nellore or Nellore × Angus steers fed different diets. Fourteen purebred and 14 crossbred steers were used, in which half of the animals of each genetic group received a diet containing whole shelled corn (WSC) and the other half whole shelled corn and sugarcane bagasse (WSCB). Stability was analyzed by the RefFinder program. To validate the selection of candidate reference genes, the expression of target genes was evaluated using the different groups of reference genes. The most stable genes were 18S, ACTB, and CASC3 for skeletal muscle; HMBS, ACTB, and 18S for the liver; and GAPDH, ACTB, and CASC3 for the jejunum, regardless of breed and diet provided. Possible errors caused in data analyses were clarified comparing the more and less stable genes as reference for normalization of the target genes FASN, ACOX, SCD1, MGAM, and SLC2A1. The use of the more stable and less stable sets of reference genes may lead to different conclusions in respect to the expression profile of the target studied gene. The selection of more suitable reference genes for each experiment is of utmost importance to ensure the reliability of gene expression studies so that they can be applied in practice.

show abstract

Expression stability of reference genes in the skeletal muscles of beef cattle

Cited by 4 publications

References 27 publications

Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

Identification of Reference Genes for Expression Studies in the Whole-Blood from Three Cattle Breeds under Two States of Livestock Weather Safety

Effects of body condition at far‐off dry period on blood biochemistry, liver triacylglycerol and muscular monocarboxylate transporter‐1 mRNA expression in tropical Holstein dairy cows during peripartum period

Reference gene selection for quantitative PCR in liver, skeletal muscle, and jejunum of Bos indicus cattle

Contact Info

Product

Resources

About