Expression And Localization Of Gnα11 And Gnαs In Sheep Skin With Different Coat Color

Natural coat color is not only one of the important biological characteristics, but also an important economic traits in woolen animals. G protein-coupled signaling pathway plays an important role in the regulation of animal coat color formation. Gnα11 and Gnas are two small downstream subunit genes of the alpha subunit of heterologous trimeric G protein. In the endothelin signaling pathway, Gnall regulates the formation of coat color through endothelin, in which endothelin and its receptor promote differentiation and maturation of melanocytes. In the Melanocortin signaling pathway, Gnas is required for the regulation of different types of pigment formation in the single melanocyte by melanin cortical hormone pathway. Therefore, Gnα11 and Gnas may be related with the formation of animal coat color.To identify the regulation effect of Gnα11 and Gnas on sheep coat pigment formation, the QRT-PCR, Western blotting, Immunohistochemistry and Immunofluorescence techniques were used to analyse the expression and location of Gnα11 and Gnas between sheep skin with different coat colors.The results showed that:1. RT-PCR successfully amplified two partial gene bands of Gnall and Gnas, the lengths of which were 300 bp and 139 bp respectively. It is indicated that Gnα11 and Gnas could normally expressed in the sheep skin.2. QRT-PCR showed that the relative expressions of Gnα11 mRNA in black and white sheep were 1.8323±0.0864 and 1.0955±0.1238 respectively, and times of which was 1.7 (P<0.01). The relative expressions of Gnas mRNA in black and white sheep were 2.8068±0.2343 and 1.1732±0.1647 respectively, and times of which was 2.4 (P<0.01). This showed that Gnal 1 and Gnas were correlated with coat color.3.Western blotting results showed the expressions of Gnall mRNA in black and white sheep were 0.4843±0.1292 and 0.2388Q0.0889 respectively, and times of which was 2.0 (P<0.05). The relative expressions of Gnas mRNA in black and white sheep were 0.4421±0.0435 and 0.2678±0.0720 respectively, and times of which was 1.7 (P<0.05).4. Immunohistochemistry and Immunofluorescence techniques showed that Gnall protein was located to outer root sheath of hair follicle in white and black sheep. Gnas protein was located to outer root sheath and dermal papilla of hair follicle in white and black sheep.In a word, the expression of Gna11 and Gnas both had differences in white and black sheep, which may be related to coat color formation of sheep.