DOI QR코드

DOI QR Code

Whole-transcriptome RNA sequencing reveals global expression dynamics and ceRNA regulatory networks related to hair follicle development and melanogenesis in goats

  • Junyin Zhao (College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science) ;
  • Jipan Zhang (College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science) ;
  • Ziyi Chen (College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science) ;
  • Min Xiao (College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science) ;
  • Yongju Zhao (College of Animal Science and Technology, Southwest University, Chongqing Key Laboratory of Herbivore Science)
  • Received : 2024.09.03
  • Accepted : 2025.03.26
  • Published : 2025.09.01

Abstract

Objective: Domestic animals, fur is a product of long-term selection by humans and the natural environment. It is generally used to distinguish between different breeds. This study aims to dissect the molecular mechanisms underlying the distinct fur characteristics of goats, particularly focusing on the molecular and regulatory differences between the Dazu Black Goat (DBG) and the Inner Mongolia Cashmere Goat (IMCG). Through whole-transcriptome analysis, we aim to identify differentially expressed RNAs and construct a ceRNA network to reveal the genetic regulation of goat hair follicle development and melanin production. Methods: Skin, hair, and cashmere samples were collected from DBG (n = 15) and IMCG (n = 17) to assess hair follicle density, length, diameter, and melanin content. Whole-transcriptome sequencing of skin tissues from DBG (n = 3) and IMCG (n = 3) identified 50,652 RNAs. Differential expression analysis was performed on mRNAs, lncRNAs, miRNAs, and circRNAs. Results: IMCG exhibited significantly higher hair follicle density, hair length, and cashmere diameter than DBG (p < 0.01), whereas DBG had significantly thicker hair and higher melanin content (p < 0.01). A total of 640 differentially expressed RNAs were identified, including 157 mRNAs, 234 lncRNAs, 72 miRNAs, and 177 circRNAs. These were enriched in pathways related to melanogenesis, hair follicle development, and GO terms such as collagen fiber organization and pigmentation. ceRNA networks constructed from differentially expressed RNAs revealed key regulatory mechanisms of coat color and hair traits. Conclusion: Whole-transcriptome sequencing revealed expression profiles and ceRNA networks involved in hair follicle development and melanogenesis in goats. These findings provide insights into the roles of coding and non-coding RNAs in fur traits, supporting future breeding strategies and textile applications.

Keywords

Acknowledgement

The authors thank the other members of Zhao's Lab who participated in the skin samples collection.

References

  1. Gong G, Fan Y, Yan X, et al. Identification of genes related to hair follicle cycle development in Inner Mongolia cashmere goat by WGCNA. Front Vet Sci 2022;9:894380. http://doi.org/10.3389/fvets.2022.894380
  2. Saleh AA, Rashad AMA, Hassanine NNAM, Sharaby MA, Zhao Y. Evaluation of morphological traits and physiological variables of several Chinese goat breeds and their crosses. Trop Anim Health Prod 2021;53:74. https://doi.org/10.1007/s11250-020-02549-3
  3. Arenas-Báez P, Torres-Hernández G, Castillo-Hernández G, et al. Coat color in local goats: influence on environmental adaptation and productivity, and use as a selection criterion. Biology 2023;12:929. https://doi.org/10.3390/biology12070929
  4. Voß K, Blaj I, Tetens JL, Thaller G, Becker D. Roan coat color in livestock. Anim Genet 2022;53:549-56. https://doi.org/10.1111/age.13240
  5. Bellone RR. Pleiotropic effects of pigmentation genes in horses. Anim Genet 2010;41:100-10. https://doi.org/10.1111/j.1365-2052.2010.02116.x
  6. Shang F, Ma R, Rong Y, et al. Construction and functional analysis of ceRNA regulatory network related to the development of secondary hair follicles in Inner Mongolia cashmere goats. Front Vet Sci 2022;9:959952. https://doi.org/10.3389/fvets.2022.959952
  7. Zhu B, Xu T, Yuan J, Guo X, Liu D. Transcriptome sequencing reveals differences between primary and secondary hair follicle-derived dermal papilla cells of the Cashmere goat (Capra hircus). PLOS ONE 2013;8:e76282. https://doi.org/10.1371/journal.pone.0076282
  8. Li J, Ba X, Li J, et al. MicroRNA-200a regulates skin pigmentation by targeting WNT5A and FZD4 in cashmere goats. Res Vet Sci 2022;147:68-73. https://doi.org/10.1016/j.rvsc.2022.03.020
  9. Wang M, Ma R, Ma Q, et al. Role of LncRNA MSTRG. 20890.1 in hair follicle development of cashmere goats. Genes 2024;15:1392. https://doi.org/10.3390/genes15111392
  10. He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2004;5:522-31. https://doi.org/10.1038/nrg1379
  11. Ma T, Li J, Li J, et al. Expression of miRNA-203 and its target gene in hair follicle cycle development of Cashmere goat. Cell Cycle 2021;20:204-10. https://doi.org/10.1080/15384101.2020.1867789
  12. Wang M, Dai H, Sheng S, et al. Discovery and functional analysis of secondary hair follicle miRNAs during annual cashmere growth. Int J Mol Sci 2023;24:1063. https://doi.org/10.3390/ijms24021063
  13. Zhu Z, Ma Y, Li Y, et al. Comparison of miRNA-101a-3p and miRNA-144a-3p regulation with the key genes of alpaca melanocyte pigmentation. BMC Mol Biol 2019;20:19. https://doi.org/10.1186/s12867-019-0137-8
  14. Li Y, Wang S, Ning J, Mao X, Ge K, Zhang R. The effects of miRNA-27a-3p on human epidermal melanocytes. Skin Res Technol 2023;29:e13345. https://doi.org/10.1111/srt.13345
  15. Mercer TR, Dinger ME, Mattick JS. Long non-coding RNAs: insights into functions. Nat Rev Genet 2009;10:155-9. https://doi.org/10.1038/nrg2521
  16. Yin RH, Wang YR, Zhao SJ, et al. LncRNA-599554 sponges miR-15a-5p to contribute inductive ability of dermal papilla cells through positive regulation of the expression of Wnt3a in cashmere goat. Electron J Biotechnol 2020;45:19-29. https://doi.org/10.1016/j.ejbt.2020.03.002
  17. Zhao W, Mazar J, Lee B, et al. The long noncoding RNA SPRIGHTLY regulates cell proliferation in primary human melanocytes. J Invest Dermatol 2016;136:819-28. https://doi.org/10.1016/j.jid.2016.01.018
  18. Wu X, Gu Y, Li S, et al. RNA-Seq reveals the roles of long noncoding RNAs (lncRNAs) in cashmere fiber production performance of cashmere goats in China. Genes 2023;14:384. https://doi.org/10.3390/genes14020384
  19. Li HM, Ma XL, Li HG. Intriguing circles: conflicts and controversies in circular RNA research. Wiley Interdiscip Rev RNA 2019;10:e1538. https://doi.org/10.1002/wrna.1538
  20. Guz M, Jeleniewicz W, Cybulski M. Interactions between circRNAs and miR-141 in cancer: from pathogenesis to diagnosis and therapy. Int J Mol Sci 2023;24:11861. https://doi.org/10.3390/ijms241411861
  21. Zhao J, Shen J, Wang Z, et al. CircRNA-0100 positively regulates the differentiation of cashmere goat SHF-SCs into hair follicle lineage via sequestering miR-153-3p to heighten the KLF5 expression. Arch Anim Breed 2022;65:55-67. https://doi.org/10.5194/aab-65-55-2022
  22. Yin RH, Zhao SJ, Jiao Q, et al. CircRNA-1926 promotes the differentiation of goat SHF stem cells into hair follicle lineage by miR-148a/b-3p/CDK19 axis. Animals 2020;10:1552. https://doi.org/10.3390/ani10091552
  23. Jiang L, Huang J, Hu Y, et al. Identification of the ceRNA networks in α-MSH-induced melanogenesis of melanocytes. Aging 2020;13:2700-26. https://doi.org/10.18632/aging.202320
  24. Salmena L, Poliseno L, Tay Y, Kats L, Pandolfi PP. A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language? Cell 2011;146:353-8. https://doi.org/10.1016/j.cell.2011.07.014
  25. Hong L, Gu T, He Y, et al. Genome-wide analysis of circular RNAs mediated ceRNA regulation in porcine embryonic muscle development. Front Cell Dev Biol 2019;7:289. https://doi.org/10.3389/fcell.2019.00289
  26. Zhang C, Yu Z, Yang S, et al. ZNF460-mediated circRPPH1 promotes TNBC progression through ITGA5-induced FAK/PI3K/AKT activation in a ceRNA manner. Mol Cancer 2024;23:33. https://doi.org/10.1186/s12943-024-01944-w
  27. Bai WL, Zhao SJ, Wang ZY, et al. LncRNAs in secondary hair follicle of cashmere goat: identification, expression, and their regulatory network in Wnt signaling pathway. Anim Biotechnol 2018;29:199-211. https://doi.org/10.1080/10495398.2017.1356731
  28. Ren H, Wang G, Chen L, et al. Genome-wide analysis of long non-coding RNAs at early stage of skin pigmentation in goats (Capra hircus). BMC Genomics 2016;17:67. https://doi.org/10.1186/s12864-016-2365-3
  29. Xu Q, Liu X, Chao Z, et al. Transcriptomic analysis of coding genes and non-coding RNAs reveals complex regulatory networks underlying the black back and white belly coat phenotype in Chinese Wuzhishan pigs. Genes 2019;10:201. https://doi.org/10.3390/genes10030201
  30. Zhang Y, Li F, Shi Y, Zhang T, Wang X. Comprehensive transcriptome analysis of hair follicle morphogenesis reveals that lncRNA-H19 promotes dermal papilla cell proliferation through the Chi-miR-214-3p/β-catenin axis in cashmere goats. Int J Mol Sci 2022;23:10006. https://doi.org/10.3390/ijms231710006
  31. Zhou S, Zeng H, Huang J, et al. Epigenetic regulation of melanogenesis. Ageing Res Rev 2021;69:101349. https://doi.org/10.1016/j.arr.2021.101349
  32. Zhang W, Wang N, Zhang T, Wang M, Ge W, Wang X. Roles of melatonin in goat hair follicle stem cell proliferation and pluripotency through regulating the Wnt signaling pathway. Front Cell Dev Biol 2021;9:686805. https://doi.org/10.3389/fcell.2021.686805
  33. Shang F, Wang Y, Ma R, et al. Expression profiling and functional analysis of circular RNAs in Inner Mongolian cashmere goat hair follicles. Front Genet 2021;12:678825. https://doi.org/10.3389/fgene.2021.678825
  34. Zhang G, Xu J, Zhang Y, Yang S, Jiang H. Expression of miRNA-1-3p and its target gene in hair follicle cycle development of Liaoning cashmere goat. Anim Biotechnol 2023;34:1937-42. https://doi.org/10.1080/10495398.2022.2058519
  35. Xiang B, Li Y, Li J, Li J, Jiang H, Zhang Q. MiR-19 3b regulated the formation of coat colors by targeting WNT10A and GNAI2 in cashmere goats. Anim Biotechnol 2023;34:796-804. https://doi.org/10.1080/10495398.2021.1998089
  36. Wu Z, Fu Y, Cao J, Yu M, Tang X, Zhao S. Identification of differentially expressed miRNAs between white and black hair follicles by RNA-sequencing in the goat (Capra hircus). Int J Mol Sci 2014;15:9531-45. https://doi.org/10.3390/ijms15069531
  37. Bai M, Wu ZZ, Huang YL, Ke J, Xu Q, Wang X. STAT3 activates the transcription of lncRNA NR2F1-AS1 to promote the progression of melanoma via regulating the miR-493-5p/GOLM1 axis. J Gene Med 2021;23:e3338. https://doi.org/10.1002/jgm.3338
  38. Li D, Zhou T, She Q, et al. Circulating exosomal miR-493-3p affects melanocyte survival and function by regulating epidermal dopamine concentration in segmental vitiligo. J Invest Dermatol 2022;142:3262-73.E11. https://doi.org/10.1016/j.jid.2022.05.1086
  39. Yamada T, Hasegawa S, Inoue Y, et al. Accelerated differentiation of melanocyte stem cells contributes to the formation of hyperpigmented maculae. Exp Dermatol 2014;23:652-8. https://doi.org/10.1111/exd.12496
  40. Fan R, Xie J, Bai J, et al. Skin transcriptome profiles associated with coat color in sheep. BMC Genomics 2013;14:389. https://doi.org/10.1186/1471-2164-14-389
  41. Bao Q, Ma X, Jia C, et al. Resequencing and signatures of selective scans point to candidate genetic variants for hair length traits in long-haired and normal-haired Tianzhu white yak. Front Genet 2022;13:798076. https://doi.org/10.3389/fgene.2022.798076
  42. Santoni G, Morelli MB, Santoni M, Nabissi M, Marinelli O, Amantini C. Targeting transient receptor potential channels by MicroRNAs drives tumor development and progression. Adv Exp Med Biol 2020;1131:605-23. https://doi.org/10.1007/978-3-030-12457-1_24
  43. Le L, Escobar IE, Ho T, et al. SLC45A2 protein stability and regulation of melanosome pH determine melanocyte pigmentation. Mol Biol Cell 2020;31:2687-702. https://doi.org/10.1091/mbc.E20-03-0200
  44. Kim H, Choi N, Kim DY, Kim SY, Song SY, Sung JH. TGF-β2 and collagen play pivotal roles in the spheroid formation and anti-aging of human dermal papilla cells. Aging 2021;13:19978-95. https://doi.org/10.18632/aging.203419
  45. Ma Y, Jin Y, Li C, Liu Y, Wang D. LncRNA MSC-AS1 motivates the development of melanoma by binding to miR-302a3p and recruiting IGF2BP2 to elevate LEF1 expression. Exp Dermatol 2021;30:1764-74. https://doi.org/10.1111/exd.14427