Abstract
Solute carrier family 7 member 11 (SLC7A11) is a cystine/glutamate exchanger, also known as xCT, has been found to play an important role in pheomelanin synthesis. Adjusting the cystine content of cells to influence pheomelanin synthesis affects the proportion of total melanin, changing mammalian coat colour. In our previous study, we used RNA-seq to show that SLC7A11 was involved in coat colour regulation in Rex rabbits. However, the precise role of SLC7A11 in rabbit coat colour formation has not been investigated. To better understand the functions of SLC7A11 in rabbits, we used RNA interference (RNAi) to explore the effects of small interfering RNA (siRNA)-mediated downregulation of SLC7A11 gene expression on the expression of melanogenesis-related genes in rabbit skin fibroblasts (RAB-9). The effects of siRNA treatment were measured by quantitative real-time polymerase chain reaction and the efficiency of RNAi was calculated. The expression levels of melanogenesis-related genes, including MITF, MC1R, Agouti, CREB1 and SLC24A5 were detected at 24 h after RNAi transfection. The results showed that MITF, MC1R, SLC24A5, Agouti and CREB1 expression was significantly downregulated after SLC7A11 inhibition. This suggested that changes in SLC7A11 gene expression could directly affect the transcription of genes related to melanin production. This provides a scientific basis for further study of the role of SLC7A11 in the formation of coat colour.
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References
Barsh G. and Cotsarelis G. 2007 How hair gets its pigment. Cell 130, 779–781.
Bauer G. L., Praetorius C., Bergsteinsdóttir K., Hallsson J. H., Gísladóttir B. K., Schepsky A. et al. 2009 The role of MITF phosphorylation sites during coat color and eye development in mice analyzed by bacterial artificial chromosome transgene rescue. Genetics 183, 581–594.
Chintala S., Li W., Lamoreux M. L., Ito S., Wakamatsu K., Sviderskaya E. V. et al. 2005 Slc7a11 gene controls production of pheomelanin pigment and proliferation of cultured cells. Proc. Natl. Acad. Sci. USA 102, 10964–10969.
Cone R. D., Lu D., Koppula S., Vage D. I., Klungland H., Boston B. et al. 1996 The melanocortin receptors: agonists, antagonists, and the hormonal control of pigmentation. Recent Prog. Horm. Res. 51, 287–317.
Conrad M. and Sato H. 2012 The oxidative stress-inducible cystine/glutamate antiporter, system XC\(^{-}\) : cystine supplier and beyond. Amino Acids 42, 231–246.
Elbashir S. M., Harborth J., Weber K. and Tuschl T. 2002 Analysis of gene function in somatic mammalian cells using small interfering RNAs. Methods 26, 199–213.
Flanagan N., Healy E., Ray A., Philips S., Todd C., Jackson I. J. et al. 2000 Pleiotropic effects of the melanocortin 1 receptor (MC1R) gene on human pigmentation. Hum. Mol. Genet. 9, 2531–2537.
Fontanesi L., Dall’Olio S., Beretti F., Portolano B. and Russo V. 2011 Coat colours in the Massese sheep breed are associated with mutations in the agouti signalling protein (ASIP) and melanocortin 1 receptor (MC1R) genes. Animal 5, 8–17.
Hannon G. J. 2002 RNA interference. Nature 418, 244–251.
Hartmeyer M., Scholzen T., Becher E., Bhardwaj R. S., Schwarz T. and Luger T. A. 1997 Human dermal microvascular endothelial cells express the melanocortin receptor type 1 and produce increased levels of IL-8 upon stimulation with alpha-melanocyte-stimulating hormone. J. Immunol. 159, 1930–1937.
Holen T., Amarzguioui M., Wiiger M. T., Babaie E. and Prydz H. 2002 Positional effects of short interfering RNAs targeting the human coagulation trigger tissue factor. Nucleic Acids Res. 30, 1757–1766.
Ito S. and Wakamatsu K. 2008 Chemistry of mixed melanogenesis–pivotal roles of dopaquinone. Photochem. Photobiol. 84, 582–592.
Lamason R. L., Mohideen M. A., Mest J. R., Wong A. C., Norton H. L., Aros M. C. et al. 2005 SLC24A5, a putative cation exchanger, affects pigmentation in zebrafish and humans. Science 310, 1782–1786.
Li H. T., He X., Zhou Z. Y., Zhao S. H., Zhang W. X., Liu G. et al. 2012 Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors. Yi Chuan. (Hereditas) 34, 1314–13149.
Maudet C. and Taberlet P. 2002 Holstein’s milk detection in cheeses inferred from melanocortin receptor 1 (MC1R) gene polymorphism. J. Dairy Sci. 85, 707–715.
Mundy N. I. and Kelly J. 2006 Investigation of the role of the agouti signaling protein gene (ASIP) in coat color evolution in primates. Mamm. Genome 17, 1205–1213.
Patel S. A., Warren B. A., Rhoderick J. F. and Bridges R. J. 2004 Differentiation of substrate and non-substrate inhibitors of transport system XC\(^{-}\) : an obligate exchanger of L-glutamate and L-cystine. Neuropharmacology 46, 273–284.
Qin L. Z., Wang W. Z., Shi L. J., Wan X. Y., Yan X. R., Weng Q. Q. et al. 2016 Transcriptome expression profiling of fur color formation in domestic rabbits using Solexa sequencing. Genet. Mol. Res. 15.
Ren Y. H., Ren B., Fan R. W., Zhu Z. W., Yang Y., Hui L. I. et al. 2009 Cloning and expression level analysis of MC1R gene in alpacas with dfferent coat color. Chinese J. Anim. Vet. Sci. 40, 841–845.
Shi L. J., Qin L. Z., Wang W. Z., Yan X. R., Weng Q. Q. and Xin-Sheng W. U. 2015 Screening candidate differentially expressed genes of Rex rabbits’ fur color with gene chips. Chinese J. Anim. Vet. Sci. 46, 568–575.
Song Y., Lin Y., Wang Y., Chen M., Deng J., Lv Q. et al. 2016 Efficient dual sgRNA-directed large gene deletion in rabbit with CRISPR/Cas9 system. Cell Mol. Life Sci. 73, 1–10.
Stoner C. J., Bininda-Emonds O. R. and Caro T. 2003 The adaptive significance of coloration in lagomorphs. Biol. J. Linn. Soc. 79, 309–328.
Xin H., Li H., Zhou Z., Zhao Z. and Wei L. 2012 Production of brown/yellow patches in the SLC7A11 transgenic sheep via testicular injection of transgene. J. Genet. Genomics 39, 281–285.
Acknowledgements
This research was financially supported by the Modern Agricultural Industrial System Special Funding (CARS-44-A-1) and Science and Technology Major Project of New Variety Breeding (Livestock and Poultry) of Zhejiang Province, China (2016C02054-10).
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Yang, N., Mu, L., Zhao, B. et al. RNAi-mediated SLC7A11 knockdown inhibits melanogenesis-related genes expression in rabbit skin fibroblasts. J Genet 97, 463–468 (2018). https://doi.org/10.1007/s12041-018-0945-5
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DOI: https://doi.org/10.1007/s12041-018-0945-5