Czech J. Anim. Sci., 2023, 68(5):189-201 | DOI: 10.17221/76/2022-CJAS

Effects of toll like receptor 4 overexpression on blood parameters, body composition, and gastrointestinal microbial diversity in sheepOriginal Paper

Xu-Ting Song1, Duo-Wei Zhao1, Qi Lu1, Jun Lu1, Mei-Yu Qi2, Ming-Hai Lu3, Xiu-Qin Yang1, Yu-Chang Yao1
1 College of Animal Science and Technology, Northeast Agricultural University, Harbin, P.R. China
2 Institute of Animal Husbandry, Heilongjiang Academy of Agricultural Sciences, Harbin, P.R. China
3 Department of Animal Science, Heilongjiang State Farms Science Technology Vocational College, Harbin, P.R. China

Genetic modifications can improve the health status and disease resistance of farm animals. In the previous study, genetically modified (GM) sheep overexpressing toll like receptor 4 (TLR4) were produced. The GM sheep had stronger ability to eliminate invasive microbes compared to the wild-type (WT) sheep. Physiological status and immune homeostasis, blood parameters, body composition, and gastrointestinal microbial diversity were evaluated in this study to elucidate the effects of TLR4 overexpression. In a set of 10 GM rams, the TLR4 mRNA and protein expression levels in the peripheral blood mononuclear cells, muscle, kidney, and spleen were higher than those of the WT (n = 10; P < 0.05). No significant differences in the parameters of red blood cells, white blood cells, platelets and in the composition of circulating T lymphocyte subsets were observed between the GM and WT sheep (P > 0.05). Of the biochemical parameters, only total protein levels differed (were higher) in the GM sheep than in the WT sheep (P < 0.05) while no significant differences were observed for the other indices (P > 0.05). GM and WT sheep showed similar offal weights and muscle fatty acid and amino acid compositions (P > 0.05). Furthermore, TLR4 overexpression did not affect the community structure of the gastrointestinal tract microbiota. Eleven KEGG pathways associated with the TLR4 gene, physiology and biochemistry, growth and metabolism, fatty acids, and amino acids did not demonstrate significant differences between the GM and WT sheep (P > 0.05). In conclusion, TLR4 overexpression had no principal or adverse effects on the physiological development and health of sheep, except for the general mRNA and protein expression levels.

Keywords: genetically modified sheep; TLR4 gene; microbiomes; immune homeostasis; biosafety assessment

Received: April 19, 2022; Accepted: April 20, 2023; Published: May 25, 2023  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Song X, Zhao D, Lu Q, Lu J, Qi M, Lu M, et al.. Effects of toll like receptor 4 overexpression on blood parameters, body composition, and gastrointestinal microbial diversity in sheep. Czech J. Anim. Sci. 2023;68(5):189-201. doi: 10.17221/76/2022-CJAS.
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    Supplementary files:

    Download file76-2022_CJAS_ESM.pdf

    File size: 574.91 kB

    Download fileSupplementary_Tables_S1-S5.xls

    File size: 116 kB

    References

    1. Bai H, Wang Z, Hu R, Kan T, Li Y, Zhang X, Zhang J, Lian L, Han H, Lian Z. A 90-day toxicology study of meat from genetically modified sheep overexpressing TLR4 in Sprague-Dawley rats. PLoS One. 2015 Apr 13;10(4): 15 p. Go to original source... Go to PubMed...
    2. Cawthraw S, Pennings JL, Hodemaekers HM, de Jonge R, Havelaar AH, Hoebee B, Johnson L, Best A, Kennedy E, La Ragione RM, Newell DG, Janssen R. Gene expression profiles induced by Salmonella infection in resistant and susceptible mice. Microbes Infect. 2011 Apr;13(4):383-93. Go to original source... Go to PubMed...
    3. Chrenek P, Ryban L, Vetr H, Makarevich AV, Uhrin P, Paleyanda RK, Binder BR. Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits. Transgenic Res. 2007 Jun;16(3):353-61. Go to original source... Go to PubMed...
    4. Chuang TH, Ulevitch RJ. Triad3A, an E3 ubiquitin-protein ligase regulating Toll-like receptors. Nat Immunol. 2004 May;5(5):495-502. Go to original source... Go to PubMed...
    5. Cloete JJ, Hoffman LC, Cloete SW. A comparison between slaughter traits and meat quality of various sheep breeds: Wool, dual-purpose and mutton. Meat Sci. 2012 Jul;91(3):318-24. Go to original source... Go to PubMed...
    6. Ekser B, Bianchi J, Ball S, Iwase H, Walters A, Ezzelarab M, Veroux M, Gridelli B, Wagner R, Ayares D, Cooper DK. Comparison of hematologic, biochemical, and coagulation parameters in α1,3-galactosyltransferase gene-knockout pigs, wild-type pigs, and four primate species. Xenotransplantation. 2012 Nov-Dec;19(6):342-54. Go to original source... Go to PubMed...
    7. Freitas VJF, Serova IA, Moura RR, Andreeva LE, Melo LM, Teixeira DIA, Pereira AF, Lopes ES Jr, Dias LPB, Nunes-Pinheiro DCS, Sousa FC, Alcantara-Neto AS, Albuquerque ES, Melo CHS, Rodrigues VHV, Batista RITP, Dvoryanchikov GA, Serov OL. The establishment of two transgenic goat lines for mammary gland hG-CSF expression. Small Rum Res. 2012;105(1-3):105-13. Go to original source...
    8. Jayachandran M, Brunn GJ, Karnicki K, Miller RS, Owen WG, Miller VM. In vivo effects of lipopolysaccharide and TLR4 on platelet production and activity: Implications for thrombotic risk. J Appl Physiol. 2007 Jan;102(1):429-33. Go to original source... Go to PubMed...
    9. Kampman-van de Hoek E, Jansman AJ, van den Borne JJ, van der Peet-Schwering CM, van Beers-Schreurs H, Gerrits WJ. Dietary amino acid deficiency reduces the utilisation of amino acids for growth in growing pigs after a period of poor health. J Nutr. 2016 Jan;146(1):51-8. Go to original source... Go to PubMed...
    10. Karsten E, Breen E, Herbert BR. Red blood cells are dynamic reservoirs of cytokines. Sci Rep. 2018 Feb 15;8(1): 12 p. Go to original source... Go to PubMed...
    11. Kelly B, Pearce EL. Amino assets: How amino acids support immunity. Cell Metab. 2020 Aug 4;32(2):154-75. Go to original source... Go to PubMed...
    12. Lee K, Uh K, Farrell K. Current progress of genome editing in livestock. Theriogenology. 2020 Jul 1;150:229-35. Go to original source... Go to PubMed...
    13. Lei Z, Zhang K, Li C, Jiao T, Wu J, Wei Y, Tian K, Li C, Tang D, Davis DI, Casper DP, Jiang H, Wang X, Wang J. Ruminal metagenomic analyses of goat data reveals potential functional microbiota by supplementation with essential oil-cobalt complexes. BMC Microbiol. 2019 Feb 4;19(1): 10 p. Go to original source... Go to PubMed...
    14. Liu J, Luo Y, Ge H, Han C, Zhang H, Wang Y, Su J, Quan F, Gao M, Zhang Y. Anti-bacterial activity of recombinant human β-defensin-3 secreted in the milk of transgenic goats produced by somatic cell nuclear transfer. PLoS One. 2013 Jun 14;8(6): 10 p. Go to original source... Go to PubMed...
    15. Lu D, Liu S, Shang S, Wu F, Wen X, Li Z, Li Y, Hu X, Zhao Y, Li Q, Li N. Production of transgenic-cloned pigs expressing large quantities of recombinant human lysozyme in milk. PLoS One. 2015 May 8;10(5): 14 p. Go to original source... Go to PubMed...
    16. Luu M, Pautz S, Kohl V, Singh R, Romero R, Lucas S, Hofmann J, Raifer H, Vachharajani N, Carrascosa LC, Lamp B, Nist A, Stiewe T, Shaul Y, Adhikary T, Zaiss MM, Lauth M, Steinhoff U, Visekruna A. The short-chain fatty acid pentanoate suppresses autoim-munity by modulating the metabolic-epigenetic crosstalk in lymphocytes. Nat Commun. 2019 Feb 15;10(1):12 p. Go to original source... Go to PubMed...
    17. Peng F, Chen X, Meng T, Li E, Zhou Y, Zhang S. Hematology and serum biochemistry parameters of captive Chinese alligators (Alligator sinensis) during the active and hibernating periods. Tissue Cell. 2018 Apr;51:8-13. Go to original source... Go to PubMed...
    18. Pisitkun P, Deane JA, Difilippantonio MJ, Tarasenko T, Satterthwaite AB, Bolland S. Autoreactive B cell responses to RNA-related antigens due to TLR7 gene duplication. Science. 2006 Jun 16;312(5780):1669-72. Go to original source... Go to PubMed...
    19. Qin J, Li Y, Cai Z, Li S, Zhu J, Zhang F, Liang S, Zhang W, Guan Y, Shen D, Peng Y, Zhang D, Jie Z, Wu W, Qin Y, Xue W, Li J, Han L, Lu D, Wu P, Dai Y, Sun X, Li Z, Tang A, Zhong S, Li X, Chen W, Xu R, Wang M, Feng Q, Gong M, Yu J, Zhang Y, Zhang M, Hansen T, Sanchez G, Raes J, Falony G, Okuda S, Almeida M, LeChatelier E, Renault P, Pons N, Batto JM, Zhang Z, Chen H, Yang R, Zheng W, Li S, Yang H, Wang J, Ehrlich SD, Nielsen R, Pedersen O, Kristiansen K, Wang J. A metagenome-wide association study of gut microbiota in type 2 diabetes. Nature. 2012 Oct 4;490(7418):55-60. Go to original source... Go to PubMed...
    20. Roy MF, Lariviere L, Wilkinson R, Tam M, Stevenson MM, Malo D. Incremental expression of Tlr4 correlates with mouse resistance to Salmonella infection and fine regulation of relevant immune genes. Genes Immun. 2006 Jul;7(5):372-83. Go to original source... Go to PubMed...
    21. Shi H, Kokoeva MV, Inouye K, Tzameli I, Yin H, Flier JS. TLR4 links innate immunity and fatty acid-induced insulin resistance. J Clin Invest. 2006 Nov;116(11):3015-25. Go to original source... Go to PubMed...
    22. Smirnov AV, Kontsevaya GV, Feofanova NA, Anisimova MV, Serova IA, Gerlinskaya LA, Battulin NR, Moshkin MP, Serov OL. Unexpected phenotypic effects of a transgene integration causing a knockout of the endogenous Contactin-5 gene in mice. Transgenic Res. 2018 Feb;27(1):1-13. Go to original source... Go to PubMed...
    23. Smith TG. Structure and function of red and white blood cells. Medicine. 2013 Apr;41(4):193-9. Go to original source...
    24. Solomon MB, Pursel VG, Paroczay EW, Bolt DJ. Lipid composition of carcass tissue from transgenic pigs expressing a bovine growth hormone gene. J Anim Sci. 1994 May;72(5):1242-6. Go to original source... Go to PubMed...
    25. Ubeda C, Lipuma L, Gobourne A, Viale A, Leiner I, Equinda M, Khanin R, Pamer EG. Familial transmission rather than defective innate immunity shapes the distinct intestinal microbiota of TLR-deficient mice. J Exp Med. 2012 Jul 30;209(8):1445-56. Go to original source... Go to PubMed...
    26. Wang S, Cao Y, Deng S, Jiang X, Wang J, Zhang X, Zhang J, Liu G, Lian Z. Overexpression of Toll-like receptor 4-linked mitogen-activated protein kinase signaling contributes to internalisation of Escherichia coli in sheep. Int J Biol Sci. 2018 Jun 3;14(9):1022-32. Go to original source... Go to PubMed...
    27. Wang S, Song X, Zhang K, Deng S, Jiao P, Qi M, Lian Z, Yao Y. Overexpression of Toll-like receptor 4 affects autophagy, oxidative stress, and inflammatory responses in monocytes of transgenic sheep. Front Cell Dev Biol. 2020 May 5;8: 14 p. Go to original source... Go to PubMed...
    28. Yang H, Zhang J, Zhang X, Shi J, Pan Y, Zhou R, Li G, Li Z, Cai G, Wu Z. CD163 knockout pigs are fully resistant to highly pathogenic porcine reproductive and respiratory syndrome virus. Antiviral Res. 2018 Mar;151:63-70. Go to original source... Go to PubMed...
    29. Yao YC, Han HB, Song XT, Deng SL, Liu YF, Lu MH, Zhang YH, Qi MY, He HJ, Wang SM, Liu GS, Li W, Lian ZX. Growth performance, reproductive traits and offspring survivability of genetically modified rams overexpressing toll-like receptor 4. Theriogenology. 2017 Jul 1;96:103-10. Go to original source... Go to PubMed...
    30. Zheng D, Liwinski T, Elinav E. Interaction between microbiota and immunity in health and disease. Cell Res. 2020 Jun;30(6):492-506. Go to original source... Go to PubMed...

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.


    Return to the content