Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Khan, Samiullah; Roberts, Juliet R.; Wu, Shu‐Biao

doi:10.1038/s41598-017-14693-2

Cited by 26 publications

(20 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To calculate the relative expression of the candidate target genes, Cq values were analysed in qbase+ software version 3.0 [52]. The Cq values of target genes were normalized against previously optimised reference genes ( TBP : TATA-Box binding protein and YWHAZ : Tyrosine 3-monooxygenase/Tryptophan 5-monooxygenase activation protein zeta) [53] to obtain normalized relative quantities of individual genes. Candidate target gene specific amplification efficiencies were used based on the method of Pfaffl [54].…”

Section: Methodsmentioning

confidence: 99%

RNA-sequencing analysis of shell gland shows differences in gene expression profile at two time-points of eggshell formation in laying chickens

Khan

Roberts

2019

BMC Genomics

Self Cite

View full text Add to dashboard Cite

BackgroundEggshell formation takes place in the shell gland of the oviduct of laying hens. The eggshell is rich in calcium and various glycoproteins synthesised in the shell gland. Although studies have identified genes involved in eggshell formation, little is known about the regulation of genes in the shell gland particularly in a temporal manner. The current study investigated the global gene expression profile of the shell gland of laying hens at different time-points of eggshell formation using RNA-Sequencing (RNA-Seq) analysis.ResultsGene expression profiles of the shell gland tissue at 5 and 15 h time-points were clearly distinct from each other. Out of the 14,334 genes assessed for differential expression in the shell gland tissue, 278 genes were significantly down-regulated (log2 fold change > 1.5; FDR < 0.05) and 413 genes were significantly up-regulated at 15 h relative to the 5 h time-point of eggshell formation. The down-regulated genes annotated to Gene Ontology (GO) terms showed anion transport, synaptic vesicle localisation, organic anion transport, secretion and signal release as the five most enriched terms. The up-regulated gene annotation showed regulation of phospholipase activities, alanine transport, transmembrane receptor protein tyrosine kinase signalling pathway, regulation of blood vessels diameter and 3, 5-cyclic nucleotide phosphodiesterase activity as the five most enriched GO terms. The putative functions of genes identified ranged from calcium binding to receptor activity. Validation of RNA-Seq results through qPCR showed a positive correlation.ConclusionsThe down-regulated genes at 15 h relative to the 5 h time-point were most likely involved in the transport of molecules and synthesis activities, initiating the formation of the eggshell. The up-regulated genes were most likely involved in calcium transportation, as well as synthesis and secretory activities of ions and molecules, reflecting the peak stage of eggshell formation. The findings in the current study improve our understanding of eggshell formation at the molecular level and provide a foundation for further studies of mRNA and possibly microRNA regulation involved in eggshell formation in the shell gland of laying hens.Electronic supplementary materialThe online version of this article (10.1186/s12864-019-5460-4) contains supplementary material, which is available to authorized users.

show abstract

Section: Methodsmentioning

confidence: 99%

RNA-sequencing analysis of shell gland shows differences in gene expression profile at two time-points of eggshell formation in laying chickens

Khan

Roberts

2019

BMC Genomics

Self Cite

View full text Add to dashboard Cite

show abstract

“…The kidney and lung tissues were collected from all chickens, and their viral RNA levels were quantified using RT-PCR. Cumulative mortality over the 5 d challenge was calculated as the number of birds that survived the challenge divided by the total number of birds challenged (Khan et al, 2017, Meir et al, 2010.…”

Section: = Smentioning

confidence: 99%

Adjuvant Effects of Platycodin D on Immune Responses to Infectious Bronchitis Vaccine in Chickens

Zhou

Mao

2020

J. Poult. Sci.

View full text Add to dashboard Cite

Adjuvants are common vaccine components. Novel adjuvants may improve the protective immunity conferred by vaccines against poultry diseases. Here, a lesshemolytic saponin, platycodin D (PD), isolated from the root of Platycodon grandiflorum was investigated as a potential alternative adjuvant. PD was tested as an adjuvant in the infectious bronchitis (IB) vaccine, because the existing IB vaccine has often failed to induce effective immune responses. The adjuvant activity of PD in conjunction with IB vaccine was evaluated in this study. Compared to control treatment, PD treatment significantly increased the proliferation of chicken peripheral blood mononuclear cells, concentration of interferon-γ in culture supernatants, and anti-IB antibody titer. In chickens pre-challenged with the Mass 41 infectious bronchitis virus (IBV), PD administration resulted in fewer and less severe clinical signs, lower mortality rate, and higher protection compared to control treatment. Histopathological examination showed that the lungs and kidneys of PD-treated chickens displayed fewer pathological lesions than those of control chickens. Our results also demonstrated that this new vaccine adjuvant improved chicken humoral and cellular immune responses without any side effects. Hence, our findings suggest that PD might serve as an effective adjuvant in IBV vaccines.

show abstract

“…The statistical significance ( P < 0.05) between mean values was determined by the Tukey-Kramer test. For gene expression data analysis, raw Cq values were analysed in qbase+ software version 3.0 against TBP and YWHAZ as reference genes [73]. The analysis was based on relative expression (2 -ΔΔCq ) using gene specific amplification efficiencies [74–76].…”

Section: Methodsmentioning

confidence: 99%

Genes involved in mitochondrial biogenesis and function may not show synchronised responses to mitochondria in shell gland of laying chickens under infectious bronchitis virus challenge

Khan

Roberts

2019

BMC Mol and Cell Biol

Self Cite

View full text Add to dashboard Cite

Background Egg formation takes place in the oviduct of laying hens over a 24 h period. Infectious bronchitis virus (IBV) causes pathological lesions in the chicken oviduct. In the current study, mitochondrial counts were determined in three different segments of the oviduct during egg formation in laying chickens challenged with IBV T strain. Nuclear DNA encoded genes that are involved in mitochondrial biogenesis, fission and function were studied in the shell gland of the oviduct undergoing virus multiplication. Results In the shell gland, the mitochondrial count was significantly lower ( P < 0.05) in the challenged group, compared with the control group. However, it did not vary in response to IBV challenge in the isthmus and magnum regions of the oviduct. The gene succinate dehydrogenase complex, subunit A, flavoprotein variant ( SDHA ) was down-regulated in the shell gland by IBV challenge ( P < 0.05), while other genes being studied did not show responses to the challenge ( P > 0.05). Differential expression of the genes was observed at different time-points of egg-shell formation. The expression levels of citrate synthase ( CS ), cytochrome C, somatic ( CYC, S ) and sodium-potassium adenosine triphosphatase ( Na + -K + ATPase ) genes were significantly higher, while those of SDHA and dynamin related protein 1 ( Drp1 ) genes were significantly lower, at 15 h compared with 5 h following oviposition of the previous egg. The expression level of peroxisome proliferator-activated receptor gamma coactivator 1-alpha ( PGC-1α ) did not show significant change at different time-points. Conclusions It was concluded that IBV T strain infection in laying hens reduced mitochondrial counts only in the shell gland region of the oviduct. The genes involved in mitochondrial biogenesis or function may not show synchronised responses to that of mitochondria in the shell gland of chickens under T strain of IBV challenge.

show abstract

Reference gene selection for gene expression study in shell gland and spleen of laying hens challenged with infectious bronchitis virus

Cited by 26 publications

References 79 publications

RNA-sequencing analysis of shell gland shows differences in gene expression profile at two time-points of eggshell formation in laying chickens

RNA-sequencing analysis of shell gland shows differences in gene expression profile at two time-points of eggshell formation in laying chickens

Adjuvant Effects of Platycodin D on Immune Responses to Infectious Bronchitis Vaccine in Chickens

Genes involved in mitochondrial biogenesis and function may not show synchronised responses to mitochondria in shell gland of laying chickens under infectious bronchitis virus challenge

Contact Info

Product

Resources

About