山羊皮肤组织miRNA测序与miR-129-5p调控黑色素生成的功能研究

doi:10.11843/j.issn.0366-6964.2024.03.015

Abstract

Abstract: This work aimed to screen key miRNA(s) that regulate skin melanin deposition in goats, and to explore its regulation mechanism on skin color or coat color. The 100-day-old fetal skin samples from the Youzhou dark goat (YZDG, n=3) and Chuandong white goat (CDWG, n=3), and skin samples from the healthy 2-3-years-old Dazu black goats (DBG, n=3), Inner Mongolia cashmere goat (IMCG, n=3) were collected to observe skin melanin deposition by section staining technology, and to explore differential miRNAs by miRNA-seq technology. Otherwise, B16-F10 cutaneous melanoma cells were cultured. The effect of miR-129-5p on melanin production was verified by cell transfection, qPCR, Western blot and melanin content detection. The results showed that melanin granules were obviously deposited in the hair bulb, hair shaft, and outer root sheath of DBG hair follicles and YZDG fetal goat skin, whereas they were not observed in the skin of CDWG fetal goats and the epidermis, hair follicles of IMCG. The sequencing results showed that 62 miRNAs were differentially expressed between YZDG and CDWG, of which 31 were up-regulated and 31 down-regulated in dark-skin goats. A total of 38 differentially expressed miRNAs were found between DBG and IMCG, 10 were up-regulated and 28 down-regulated in black-coated goats. The result showed miR-129-5p was significantly highly expressed in dark-skin and black-coated goats (P<0.05). After over-expression of miR-129-5p, the cell melanin deposition in the mimics group was increased by 18.9% (P<0.05), the expression of the TYR and TYRP1 genes was up-regulated by 57.3% and 16.5%, respectively (P<0.05), and the protein expression was significantly up-regulated by 49.2% and 40.2% (P<0.05); but the mRNA and protein expressions of MITF were not significantly changed (P>0.05). After inhibition of miR-129-5p, the mRNA expression of TYR gene and its protein expression level in the inhibitor group were significantly down-regulated by 38.9% and 21.1%, respectively (P<0.05). And TYRP1, MITF protein expression levels were down-regulated by 25.3% and 28.4%, respectively (P<0.05). The results indicated that the miR-129-5p was differentially expressed in different skin colors or coat colors of goats, and it could affect melanin deposition by regulating the transcript and protein abundance of melanin-related genes (such as TYR and TYRP1). These results demonstrate that miR-129-5p is an important regulator involved in the skin melanogenesis of goats.

Key words: Capra hircus, skin color, miR-129-5p, melanogenesis, TYR

CLC Number:

S827.2

XIAO Min, ZHAO Wei, SUN Wu, NA Risu, ZHAO Le, LIU Taolu, ZHANG Jipan, ZHAO Yongju. The Skin Tissue miRNA-seq and Regulation Mechanism of miR-129-5p on Melanogenesis in Goat (Capra hircus)[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1019-1029.

References

[1] 常洪.山羊的毛色遗传[J].西安联合大学学报, 1999, 2(2):1-4.
CHANG H.Genetics of hair color in goats[J].Journal of Xi'an United University, 1999, 2(2):1-4.(in Chinese)
[2] 杜小龙, 王麒, 张乐超, 等.山羊DCT基因启动子区甲基化水平、SNP与毛色特征关系研究[J].畜牧兽医学报, 2019, 50(2):271-279.
DU X L, WANG Q, ZHANG L C, et al.Study on the relationship between methylation level, SNP in promoter region of DCT gene and hair color in goat[J].Acta Veterinaria et Zootechnica Sinica, 2019, 50(2):271-279.(in Chinese)
[3] LIANG D, ZHAO P J, SI J F, et al.Genomic analysis revealed a convergent evolution of LINE-1 in coat color:a case study in water buffaloes (Bubalus bubalis)[J].Mol Biol Evol, 2021, 38(3):1122-1136.
[4] LI M N, KNAPP S K, IDEN S.Mechanisms of melanocyte polarity and differentiation:What can we learn from other neuroectoderm-derived lineages?[J].Curr Opin Cell Biol, 2020, 67:99-108.
[5] KIM J Y, KIM J, AHN Y, et al.Autophagy induction can regulate skin pigmentation by causing melanosome degradation in keratinocytes and melanocytes[J].Pigment Cell Melanoma Res, 2020, 33(3):403-415.
[6] CAO W, ZHOU X H, MCCALLUM N C, et al.Unraveling the Structure and Function of Melanin through Synthesis[J].J Am Chem Soc, 2021, 143(7):2622-2637.
[7] BRENNER M, HEARING V J.The protective role of melanin against UV damage in human skin[J].Photochem Photobiol, 2008, 84(3):539-549.
[8] SCHALLREUTER K U, KOTHARI S, CHAVAN B, et al.Regulation of melanogenesis——controversies and new concepts[J].Exp Dermatol, 2008, 17(5):395-404.
[9] 章誉兴, 吴宏, 于黎.哺乳动物毛色调控机制及其适应性进化研究进展[J].遗传, 2021, 43(2):118-133.
ZHANG Y X, WU H, YU L.Progress on coat color regulation mechanism and its association with the adaptive evolution in mammals[J].Hereditas (Beijing), 2021, 43(2):118-133.(in Chinese)
[10] JIA X B, DING P, CHEN S Y, et al.Analysis of MC1R, MITF, TYR, TYRP1, and MLPH genes polymorphism in four rabbit breeds with different coat colors[J].Animals (Basel), 2021, 11(1):81.
[11] LAI X L, WICHERS H J, SOLER-LOPEZ M, et al.Structure and function of human tyrosinase and tyrosinase-related proteins[J].Chem Eur J, 2018, 24(1):47-55.
[12] ZHOU S H, ZENG H L, HUANG J H, et al.Epigenetic regulation of melanogenesis[J].Ageing Res Rev, 2021, 69:101349.
[13] BAUER G L, PRAETORIUS C, BERGSTEINSDÓTTIR K, et al.The role of MITF phosphorylation sites during coat color and eye development in mice analyzed by bacterial artificial chromosome transgene rescue[J].Genetics, 2009, 183(2):581-594.
[14] CHEN T Z, ZHAO B L, LIU Y, et al.MITF-M regulates melanogenesis in mouse melanocytes[J].J Dermatol Sci, 2018, 90(3):253-262.
[15] D'MELLO S A N, FINLAY G J, BAGULEY B C, et al.Signaling pathways in melanogenesis[J].Int J Mol Sci, 2016, 17(7):1144.
[16] TIAN X, JIANG J B, FAN R W, et al.Identification and characterization of microRNAs in white and brown alpaca skin[J].BMC Genomics, 2012, 13:555.
[17] WU Z Y, FU Y H, CAO J H, et al.Identification of differentially expressed miRNAs between white and black hair follicles by RNA-sequencing in the goat (Capra hircus)[J].Int J Mol Sci, 2014, 15(6):9531-9545.
[18] WANG X C, WU Y F, DU P, et al.Study on the mechanism of miR-125b-5p affecting melanocyte biological behavior and melanogenesis in vitiligo through regulation of MITF[J].Dis Markers, 2022, 2022:6832680.
[19] WU D T, CHEN J S, CHANG D C, et al.Mir-434-5p mediates skin whitening and lightening[J].Clin Cosmet Investig Dermatol, 2008, 1:19-35.
[20] DONG C S, WANG H D, XUE L L, et al.Coat color determination by miR-137 mediated down-regulation of microphthalmia-associated transcription factor in a mouse model[J].RNA, 2012, 18(9):1679-1686.
[21] DJEBALI S, DAVIS C A, MERKEL A, et al.Landscape of transcription in human cells[J].Nature, 2012, 489(7414):101-108.
[22] SHI X L, WU J P, LANG X, et al.Comparative transcriptome and histological analyses provide insights into the skin pigmentation in Minxian black fur sheep (Ovis aries)[J].PeerJ, 2021, 9:e11122.
[23] YUAN W, QIN H, BI H, et al.Ssc-mir-221-3p regulates melanin production in Xiang pigs melanocytes by targeting the TYRP1 gene[J].BMC Genomics, 2023, 24(1):369.
[24] 唐朋, 凌笑笑, 高泽成, 等.不同毛色牦牛皮肤组织学观察及MC1R基因功能验证[J].畜牧兽医学报, 2018, 49(7):1377-1386.
TANG P, LIN X X, GAO Z C, et al.Skin histological observation of yak with different coat colors and MC1R functional verification[J].Acta Veterinaria et Zootechnica Sinica, 2018, 49(7):1377-1386.(in Chinese)
[25] HUSHCHA Y, BLO I, OTON-GONZALEZ L, et al.microRNAs in the regulation of melanogenesis[J].Int J Mol Sci, 2021; 22(11):6104.
[26] TANG P, ZHOU J J, LIU H G, et al.Depletion of lncRNA MEG3 ameliorates imatinib-induced injury of cardiomyocytes via regulating miR-129-5p/HMGB1 axis[J].Anal Cell Pathol (Amst), 2023, 2023:1108280.
[27] YAO F, ZHAO B H, HU S S, et al.miR-129-5p participates in hair follicle growth by targeting HOXC13 in rabbit[J].Genes (Basel), 2022, 13(4):679.
[28] YIN C, TIAN Y, YU Y, et al.miR-129-5p inhibits bone formation through TCF4[J].Front Cell Dev Biol, 2020, 8:600641.
[29] MO X, KAZMI H R, PRESTON-ALP S, et al.Interferon-gamma induces melanogenesis via post-translational regulation of tyrosinase[J].Pigment Cell Melanoma Res, 2022, 35(3):342-355.
[30] HELSING P, NYMOEN D A, ROOTWELT H, et al.MC1R, ASIP, TYR, and TYRP1 gene variants in a population-based series of multiple primary melanomas[J].Genes Chromosomes Cancer, 2012, 51(7):654-661.
[31] 赵若阳, 李超, 图格琴, 等.越背花毛蒙古马皮肤组织中色素生成相关基因TYR、TYRP1及DCT的表达分析[J].农业生物技术学报, 2019, 27(6):1042-1050.
ZHAO R Y, LI C, TU G Q, et al.Expression analysis of melanogenesis related genes TYR, TYRP1 and DCT in Tobiano Mongolian Horse (Equus caballus) skin tissue[J].Journal of Agricultural Biotechnology, 2019, 27(6):1042-1050.(in Chinese)
[32] ZHAO Y Y, WANG P C, MENG J Z, et al.MicroRNA-27a-3p Inhibits Melanogenesis in Mouse Skin Melanocytes by Targeting Wnt3a[J].Int J Mol Sci, 2015, 16(5):10921-10933.
[33] 张利环, 马悦悦, 刘文艳, 等.microRNA-96-5p靶向调控羊驼黑色素细胞中MITF基因的表达[J].畜牧兽医学报, 2020, 51(6):1229-1237.
ZHANG L H, MA Y Y, LIU W Y, et al.microRNA-96-5p targets MITF gene in Alpaca melanocytes[J].Acta Veterinaria et Zootechnica Sinica, 2020, 51(6):1229-1237.(in Chinese)
[34] ZHU Z W, MA Y Y, LI Y, et al.Comparison of miRNA-101a-3p and miRNA-144a-3p regulation with the key genes of alpaca melanocyte pigmentation[J].BMC Mol Biol, 2019, 20(1):19.
[35] WANG P C, ZHAO Y Y, FAN R W, et al.MicroRNA-21a-5p functions on the regulation of melanogenesis by targeting Sox5 in mouse skin melanocytes[J].Int J Mol Sci, 2016, 17(7):959.
[36] VASUDEVAN S, TONG Y C, STEITZ J A.Cell-cycle control of microRNA-mediated translation regulation[J].Cell Cycle, 2008, 7(11):1545-1549.
[37] ØROM U A, NIELSEN F C, LUND A H.MicroRNA-10a binds the 5'UTR of ribosomal protein mRNAs and enhances their translation[J].Mol Cell, 2008, 30(4):460-471.
[38] GAGNON K T, LI L D, CHU Y J, et al.RNAi factors are present and active in human cell nuclei[J].Cell Rep, 2014, 6(1):211-221.
[39] MAJID S, DAR A A, SAINI S, et al.MicroRNA-205-directed transcriptional activation of tumor suppressor genes in prostate cancer[J].Cancer, 2010, 116(24):5637-5649.
[40] XIAO M, LI J, LI W, et al.MicroRNAs activate gene transcription epigenetically as an enhancer trigger[J].RNA Biol, 2017, 14(10):1326-1334.

The Skin Tissue miRNA-seq and Regulation Mechanism of miR-129-5p on Melanogenesis in Goat (Capra hircus)

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 6

Recommended Articles

Metrics

Comments

[1]	LIU Hang, WANG Huanhuan, GE Ying, ZHANG Lei, ZHANG Weiwu, WEI Yinghui, LI Qinghai, FAN Jinghui, ZHANG Xuedong. Screening of Candidate Genes of Skin Color of Black-Bone Chicken Based on Transcriptome and Proteome [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2320-2329.
[2]	SONG Xingchao, LIU Linling, PAN Hongjun, ZHAO Jiaping, JIA Yun, YANG Fuhe, XU Chao. Cloning,SNPs Screening and mRNA Differential Expression Analysis of TYR Gene in Skin of Mink(Neovison vison) [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 66-76.
[3]	XU Dongmei, ZHU Zhiwei, TANG Zhongwei, LI Pengfei, DONG Changsheng, ZHAO Yujun. Effect of Interaction between miR-186-5p and α-MSH on Melanogenesis in Sheep Melanocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 19-27.
[4]	TANG Peng, LING Xiao-xiao, GAO Ze-cheng, JIA Cong-jun, LIANG Chun-nian, WU Xiao-yun, CHU Min, YAN Ping. Skin Histological Observation of Yak with Different Coat Colors and MC1R Functional Verification [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(7): 1377-1386.
[5]	WU Xiao-qian，LIU Chen-dong，DU Jing-jing，LUO Jia，ZHU Li，ZHANG Shun-hua. Research Progress of the Role of microRNAs in the Regulation of Animal Coat and Skin Color [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(6): 1086-1092.
[6]	WANG Huan-huan，CHEN Mei-ling，LOU Li-feng，ZHANG Lei，ZHANG Cheng-xian，CHEN Xian-hui，ZHANG Xue-dong. GNAS Gene Promoter Mutation in Chicken and the Correlation with Skin Color Traits [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2354-2361.