Relationship between MC1R c.676A>G and goat skin pigmentation and its effect on melanin synthesis

doi:10.3969/j.issn.1004-1524.2022.12.06

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (12): 2629-2639.DOI: 10.3969/j.issn.1004-1524.2022.12.06

• Animal Science • Previous Articles Next Articles

Relationship between MC1R c.676A>G and goat skin pigmentation and its effect on melanin synthesis

JIANG Jing¹^,²(), REN Hangxing¹^,², ZHOU Peng¹^,², SUN Xiaoyan¹^,², LI Jie¹^,², FU Lin¹^,², ZHANG Li¹^,², LIU Liangjia¹^,², WANG Gaofu¹^,²^,^*()

1. Chongqing Academy of Animal Sciences, Rongchang 402460, Chongqing, China
2. Chongqing Engineering Research Center for Goat, Rongchang 402460, Chongqing, China

Received:2022-01-12 Online:2022-12-25 Published:2022-12-26
Contact: WANG Gaofu

Abstract

Abstract:

In order to reveal the relationship between goat MC1R gene c.676A>G mutation and goat skin pigmentation, and the effect of mutation on melanin synthesis and the expression of pigment-related genes in the downstream of the cAMP signaling pathway: on the one hand, the direct sequencing was used to detect the MC1R c.676A>G mutation of Youzhou dark-skin goats and the hybrid offsprings (120 individuals), and the abdominal skin color of individuals of different genotypes was determined with the colorimeter, and the relationship between c.676A>G mutation and skin pigmentation was analyzed; on the other hand, the eukaryotic expression vectors of wild type (MC1R c.676A) and mutant type (MC1R c.676G) were constructed and transfected into B16-F10 cells for overexpression by liposome-mediated method, and the differences of melanin in each group were detected by the microplate reader and the expression differences of pigment-related genes in the downstream of the cAMP signaling pathway (MITF, TYR, TYRP1, DCT) in cells were detected by quantitative real-time PCR. The results showed that the c.676A>G site of Youzhou dark-skin goats and their hybrid offsprings was dominated by the GG genotype, and the skin color values of different genotype populations did not show significant difference (P>0.05). The melanin in mutant group cells (MC1R c.676G) showed significantly (P<0.01) higher value than that of the wild group cells (MC1R c.676A). The relative expression of MITF, TYRP1 and DCT in the mutant group cells was significantly (P<0.01) higher than that of the wild group, and the relative expression of TYR in the mutant group was significantly (P<0.05) higher than that of the wild group. In summary, MC1R c.676A>G did not directly affect goat skin pigmentation. However, MC1R c.676G could promote the synthesis of melanin in B16-F10 cells and could increase the expression of MITF, TYR, TYRP1 and DCT genes in the downstream of cAMP signaling pathway, indicating that MC1R c.676A>G was closely related to melanin synthesis.

Key words: goats, MC1R gene, mutation, melanin synthesis, cAMP signaling pathway

CLC Number:

S827

JIANG Jing, REN Hangxing, ZHOU Peng, SUN Xiaoyan, LI Jie, FU Lin, ZHANG Li, LIU Liangjia, WANG Gaofu. Relationship between MC1R c.676A>G and goat skin pigmentation and its effect on melanin synthesis[J]. Acta Agriculturae Zhejiangensis, 2022, 34(12): 2629-2639.

Figures/Tables 8

Table 1 Parameters of primers used in experiment

引物名称 Primer name	正向引物序列 Forward primer sequences(5'→3')	反向引物序列 Reverse primer sequences(5'→3')	产物大小 Product size/bp
Goat-K-MC1R	TGAGAGCAAGCACCCTTTCCT	GGTCTAGCGATCCTCTTTGTCA	1 062
Goat-MC1R	CTCCATCTTCTACGCCCTGC	GGACGACCGTGTGGTTGTAG	134
Mouse-ACTB	TCCAGCCTTCCTTCTTGGGTAT	GTTGGCATAGAGGTCTTTACGG	104
Mouse-MITF	CAACGGGAACAGCAACGAGC	TTCACCAGATCAGGCGAGCA	164
Mouse-TYR	AATCCTAACTTACTCAGCCCAGC	TAATAGTGGTCCCTCAGGTGTTC	120
Mouse-TYRP1	ACTGATGCGGTCTTTGACGAATG	GCCTGTTATGTCCAATAGGTGCG	91
Mouse-DCT	TCTTCCTAACCGCAGAGCAACTT	GATTCCAATGACCACTGAGAGAG	101

Table 2 Genotype (allele) frequency and genetic characteristics of MC1R gene c.676A>G locus in Youzhou dark-skin goats and its hybrid progeny

群体 Group									基因型及其频率 Genotype and frequency
酉州乌羊Youzhou dark-skin goats	0.709	0.291	1.410	0.248	0.617	0.432	A	0.188	AA	0.059(1)
									AG	0.235(4)
							G	0.812	GG	0.706(12)
杂交后代Hybrid offsprings	0.751	0.248	1.331	0.217	4.971	0.025	A	0.145	AA	0.049(5)
									AG	0.194(20)
							G	0.854	GG	0.757(78)

Table 3 Skin color of Youzhou dark-skin goats and their hybrid offsprings under different MC1R gene c.676A>G locus polymorphism

基因型 Genotype	酉州乌羊 Youzhou dark-skin goats	杂交后代 Hybrid offsprings
AA	70.07±12.51	31.48±5.59
AG	79.67±6.25	29.92±2.79
GG	75.68±3.61	27.14±1.41

Fig.1 Cell morphology of each group at different time A, B, C, D represent BC group, NC group, MC1R-A group and MC1R-G group, respectively. 1, 2 and 3 represent pre-transfection, 24 h after transfection and 72 h after differentiation, respectively.

Fig.2 Results of cell transfection in each group at different time A, B, C, D represent BC group, NC group, MC1R-A group and MC1R-G group, respectively. 1 and 2 represent 24 h after transfection and 72 h after differentiation, respectively.

Fig.3 Relative expression level of MC1R gene in each group at 72 h of differentiation Bars marked without the same uppercase letters indicate significant difference at P<0.01. The same as below.

Fig.4 Percentage of melanogenesis in each group Bars marked without the same lowercase letters indicate significant difference at P<0.05. The same as below.

Fig.5 Expression level of cytochrome related genes in each group

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Relationship between MC1R c.676A>G and goat skin pigmentation and its effect on melanin synthesis

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