Expression of Serine Biosynthesis Pathway Genes in Breast Muscles of Iranian Native Turkeys with Divergent Feed Efficiency

پژوهشهای علوم دامی ایران

Article 9, Volume 14, Issue 3 - Serial Number 51, September 2022, Pages 427-437   

Document Type: Research Articles

DOI: 10.22067/ijasr.2022.74768.1061

Authors

Zahra Pezeshkian¹; Seyed Ziaeddin Mirhoseini^{* 2}; Shahrokh Ghovvati¹; Esmaeil Ebrahimie³

¹Department of Animal Sciences, University of Guilan, Rasht, Guilan, Iran.

²Department of Anmal Sciences- University of Guilan, Rasht,Guilan,Iran

³Department of Agriculture and Plant Breeding, Shiraz University, Shiraz, shiraz, Iran.

Abstract

Introduction: Feed is the main cost part of poultry production. High feed efficiency poultry produce less feed and less excrement per unit weight gain. Therefore, a comprehensive understanding of the biological mechanisms that control feed efficiency is crucial for the development of optimal breeding and selection strategies. The serine biosynthesis pathway is one of the most important pathways in animals with high feed efficiency. The aim of this study was to investigate the expression of PHGDH, PSAT1 and PSPH genes by real-time PCR in Iranian native turkeys with high and low feed efficiencies.
Materials and Methods: Iranian native male turkeys (n=500) were reared up to 20 weeks of age under standard production guidelines. Then 75 turkeys were randomly selected and placed in separate cages with free access to water and feed from 20 to 24 weeks. Turkeys were ranked based on feed conversion ratio (FCR) and three turkeys with the highest and three turkeys with lowest feed efficiency were selected as high feed efficiency (HFE) and low feed efficiency (LFE) birds, respectively. After slaughter of turkeys, RNA was extracted from breast tissue. Quantity and purity of the extracted RNAs were determined using a nanodrop device and its quality was evaluated using 1% agarose gel electrophoresis. Sequences of PSPH, PHGDH, PSAT1 and RSP7 genes were collected from the NCBI database. The primer was designed using Primer Premier version 5 software. All primers were synthesized by Sinaclon (Iran). In this study, RSP7 gene was used as a reference gene. Then, cDNA synthesis was performed. The best amplification temperature for simultaneous amplification of target and reference genes was determined. Samples were amplified for each gene with 3 replications using real-time PCR reaction. Significance level between treatments for each gene was determined separately using t-test in SAS software version 9.2 (P<0.05).
Results and Discussion: Results of ultraviolet light absorption measurements at 260 and 280 nm by the nanodrop device showed that the quantity and quality of RNA extracted from the breast muscle samples were of high purity and not contaminated. The range of RNA concentration of the extracted samples was between 480 to 962 ng/μl and the ratio of absorption at 260 and 280 wavelength was about 2.1, which indicates the good quality of the extracted RNAs. The most suitable temperature was selected for specific binding of primers and simultaneous amplification of target genes and temperature control of 58 °C. To investigate and confirm the specificity of replication, melting curves were created to ensure the specificity of the amplified products, the absence of non-specific bands and secondary structures such as hairpin and primer-dimer structures. The results showed that there was only one narrow peak for each gene. The results of studying the expression of serine biosynthesis pathway genes (PSPH, PHGDH and PSAT1) showed that the expression level of these genes in HFE male turkeys was significantly higher than LFE male. Higher expression of PSPH, PHGDH and PSAT1 genes in HFE animals than in LFE animals indicates activation of the serine amino acid biosynthesis pathway, which itself can provide precursors for the Krebs cycle and purine biosynthesis. Glucose is the main source of metabolic energy in the body. When glucose enters the cell, glycolysis begins in the cytoplasm. The pathway of glycolysis and Glutamine catabolism produces an intermediate metabolite called 3-phosphoglycerate, which is gradually catalyzed to serine by PHGDH, PSAT1, and PSPH. Eventually serine is converted to glycine. Activation of this pathway indicates the higher ability of HFE animals to make better use of energy sources such as glucose, which increases protein production in breast muscle tissue and enhances volume and weight of muscle tissue in HFE turkeys.
Conclusion: The results of this study showed that the expression of serine biosynthesis pathway genes (PSPH, PHGDH and PSAT1) was significantly higher in high feed efficiency turkeys than in low feed efficiency turkeys. In fact, these results at the level of molecular biology show that turkeys with higher feed efficiency cultivate better use of energy received from feed. Activation of this pathway increases the biosynthesis of various amino acids and thus increases protein and muscle mass in birds. The results of this study can be a promising window to introduce genes that affect feed efficiency in order to further investigate the population and larger flocks of birds.

Keywords

Feed efficiency; Gene expression; Real-time PCR; Serine biosynthesis; Turkey

References

1. Aggrey, S. E., Karnuah, A. B., Sebastian, B., & Anthony, N. B. (2010). Genetic properties of feed efficiency parameters in meat-type chickens. Genetic Selection Evolution, 42, 25. https://doi.org/1186/1297-9686-42-25
2. Artegoitia, V. M., Foote, A. P., Lewis, R. M., & Freetly, H. C. (2019). Metabolomics profile and targeted lipidomics in multiple tissues associated with feed efficiency in beef steers. ACS Omega, 4(2), 3973-3982. https://doi.org/1021/acsomega.8b02494
3. Bottje, W., & Carstens, G. (2009). Association of mitochondrial function and feed efficiency in poultry and livestock species. Journal of Animal Science, 87(suppl_14), E48-E63. https://doi.org/2527/jas.2008-1379
4. Case, L. A., Wood, B. J., & Miller, S. P. (2012). The genetic parameters of feed efficiency and its component traits in the turkey (Meleagris gallopavo). Genetic Selection Evolution, 44, 2. https://doi.org/1186/1297-9686-44-2
5. Ebrahimzadeh-Allahabad, A., Mahmodian, Z., Pezeshkian, Z., & Mollaei, A. (2015). Estimation of genetic and phenotypic parameters of some important economic traits in Khazak native hen in Iran. Animal Genetic Resources/Resources Génétiques Animales/Recursos Genéticos Animales, 57: 99-103. https://doi.org/1017/S2078633615000272
6. Fonseca, L. D., Eler, J. P., Pereira, M. A., Rosa, A. F., Alexandre, P. A., Moncau, C. T., Salvato, F., Fernandes, L. R., Palmisano, G., Ferraz, J. B. S., & Fukumasu, H. (2019). Liver proteomics unravel the metabolic pathways related to feed efficiency in beef cattle. Scientific Reports, 9(1), 1-11. https://doi.org/1038/s41598-019-41813-x
7. Gong, S. (2010). Identification of nover growth signalling pathway in sheep skeletal muscle by comparative microarray analysis. Doctoral dissertation. University of Nottingham. The UK.
8. Kong, B. W., Song, J., Lee, J., Hargis, B., Wing, T., Lassiter, K., & Bottje, W. (2011). Gene expression in breast muscle associated with feed efficiency in a single male broiler line using a chicken 44K oligo microarray. I. Top differentially expressed genes. Poultry Science, 90(11), 2535-2547. https://doi.org/3382/ps.2011-01435
9. Pan, S., Fan, M., Liu, Z., Li, X., & Wang, H. (2021). Serine, glycine and one‑carbon metabolism in cancer. International Journal of Oncology, 58(2), 158-170. https://doi.org/3892/ijo.2020.5158
10. Parr, T., Al-Doski, S., Hemmings, K., Daniel, Z., Brown, D., Lu, C., Hodgman,, May, S., & Brameld, J. (2015). Increased expression of serine biosynthetic pathway genes is associated with skeletal muscle hypertrophy in sheep. Proceedings of the Nutrition Society, 74(OCE2). https://doi.org/10.1017/S0029665115002013
11. Pfaffl, M. W. (2001). A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Research, 29(9): e45-e45. https://doi.org/1093/nar/29.9.e45
12. Pirani, N., Elyasi Zarringhabaei, G. H., & Taghizadeh, A. (2010). Genetic relationship of 6 Iranian native chicken populations using RAPD markers. Animal Science Researches, 19(1), 69-79.
13. Richards, M. (2003). Genetic regulation of feed intake and energy balance in poultry. Poultry science, 82(6), 907-916. https://doi.org/1093/ps/82.6.907
14. Salahi, A. (2014). A review of the turkey meat production industry in Iran. Zootec International, 36, 24-29.
15. Wang, X., & Kadarmideen, H. N. (2019). Metabolomics analyses in high-low feed efficient dairy cows reveal novel biochemical mechanisms and predictive biomarkers. Metabolites, 9(7): 151. https://doi.org/3390/metabo9070151
16. Willems, O. W. (2014). Evaluation methods and technologies for improving feed efficiency in the turkey (Meleagris gallopavo). Doctoral dissertation. University of Guelph. Guelph. Canada.
17. Willems, O. W., Miller, S. P., & Wood, B. J. (2013). Assessment of residual body weight gain and residual intake and body weight gain as feed efficiency traits in the turkey (Meleagris gallopavo). Genetics Selection Evolution, 45(1), 26. https://doi.org/1186/1297-9686-45-26
18. Xu, C., Wang, X., Zhuang, Z., Wu, J., Zhou, Sh., Quan, J., Ding, R., Ye, Y., Peng, L., Wu, Z., Zheng, E., & Yang, J. (2020). A transcriptome analysis reveals that hepatic glycolysis and lipid synthesis are negatively associated with feed efficiency in DLY pigs. Scientific Reports, 10(1), 1-12. https://doi.org/1038/s41598-020-66988-6
19. Yang, H., Huang, X., Fang, S., He, M., Zhao, Y., Wu, Z., Yang, M., Zhang, Zh., Chen, C., &Huang, L. (2017). Unraveling the fecal microbiota and metagenomic functional capacity associated with feed efficiency in pigs. Frontiers in Microbiology, 8, 1555. https://doi.org/3389/fmicb.2017.01555
20. Zhou, N., Lee, W. R., & Abasht, B. (2015). Messenger RNA sequencing and pathway analysis provide novel insights into the biological basis of chickens’ feed efficiency. BMC Genomics, 16(1), 1-20. https://doi.org/1186/s12864-015-1364-0