敲降TYRP1基因对香猪表皮黑素细胞黑色素生成的影响

doi:10.11843/j.issn.0366-6964.2023.10.013

Abstract

Abstract: The aim of this study was to explore the effect of tyrosine-related protease 1 (tyrosinase related protein 1, TYRP1) on melanin production in skin melanocytes of Xiang pigs. Three 3-month-old healthy pigs with the hair color characteristics of "two-head-black" animals were selected. The black skin tissue on the head and white skin tissue on the back were collected for each pig.The structure of hair follicles and the distribution characteristics of melanocytes were observed by HE staining; The epidermal melanocytes were cultured in vitro and identified by Dopa (L-Dopa) staining, quantitative real-time PCR and Western Blot methods; Five siRNAs were constructed from TYRP1 gene sequence, and the siRNA with the highest interference efficiency was transfected by quantitative real-time PCR. After successful interference of TYRP1 in melanocytes, the mRNA and protein expression levels of TYRP, TYRP1 and TYRP2 were measured by quantitative real-time PCR and Western Blot, the total intracellular melanin content also been measured. The results showed that intact hair follicle structures were observed in both the black and white skin of Xiang pigs, and the melanocytes were mainly distributed in the outer root sheath of hair follicles and epidermes in the black hairy skin of Xiang pig. Under the treatment of MelM medium, the melanocytes grew vigorously, and the results of cell morphology, growth curve, Dopa staining, quantitative real-time PCR and Western Blot all indicated that the cultured melanocytes maintained normal biological characteristics. Knocking down of TYRP1 downregulated the mRNA and protein expression of melanogenesis-related genes TYR, TYRP1 and TYRP2 in Xiang pig epidermal melanocytes, and also had an inhibitory effect on melanin formation. The TYRP1 gene was able to affect the melanin production in melanocyte epidermal cells,the results provide experimental reference and basic data to explore the molecular mechanism of TYRP1 gene on melanin deposition in Xiang pigs.

Key words: TYRP1 gene, melanocytes, siRNA, HE staining, L-Dopa staining, melanin

CLC Number:

S828.2

BI Huan, QIN Hai, YUAN Wei, ZHANG Yudan, ZHANG Yiyu, CHEN Wei. Effect of Knocking down TYRP1 Gene on Melanin Production in the Xiang Pig Epidermal Melanocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4143-4153.

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Effect of Knocking down TYRP1 Gene on Melanin Production in the Xiang Pig Epidermal Melanocytes

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