不同毛色牦牛皮肤组织学观察及MC1R基因功能验证

doi:10.11843/j.issn.0366-6964.2018.07.006

Abstract

Abstract:

The objectives of this study were to elucidate the characteristics of skin histomorphology in yaks with different coat colors and to discover the potential molecular mechanism by which MC1R induced melanin synthesis. Skin tissues were collected from the hindquarters of 6 Datong yaks(dark brown) and 6 Tianzhu yaks(white). Toluidine blue staining was used to reveal the content and distribution of both melanin and mature melanocytes in the skin tissues of different coat colors. MC1R expression was determined by quantitative real-time polymerase chain reaction(qRT-PCR). Afterward,shRNA interference vectors for MC1R were transfected into B16 murine melanoma cells,and transfection efficiency was quantified by qRT-PCR and Western blot assay. The expressions of hair color-related genes(Agouti,MITF and TYR) in each group were analyzed and compared,and melanin production was detected with a microplate reader at different time points. The results showed that a large number of melanocytes distributed around the epidermis and hair follicles of both types of yaks. Tiny amounts of melanin were detected at the base of the epidermis and the hair roots of Tianzhu white yak skin tissues,while large amounts of melanin were discovered in the epidermis and hair follicles of Datong yak skin tissues. The expression of MC1R in Tianzhu white yaks was significantly lower than that in Datong yaks(P<0.01).In B16 cells with successful inhibition of MC1R,expression of TYR was very significantly lower(P<0.01),MITF expression was significantly lower(P<0.05),Agouti expression was not significantly different(P>0.05),and melanin content was lower than in cells that expressed MC1R. These results indicate that MC1R plays an important role in melanin synthesis and formation of coat color. To summarize,shRNA MC1R vectors were successfully transfected into B16 mouse melanoma cells;inhibition of MC1R in B16 cells was consistent with reduced expression of the downstream gene TYR,which lowered the melanin content in skin tissues. Therefore,we conclude that massive deposition of melanin and high expression of MC1R are responsible for the differences in yak coat colors.

Key words: yak, skin tissue morphology, shRNA MC1R, hair color-related genes, melanogenesis

CLC Number:

S823.8⁺5.2

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.

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Skin Histological Observation of Yak with Different Coat Colors and MC1R Functional Verification

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