Study On The Expression Characteristics Of TRPM1 In Different Coat Color Sheep Skin At Different Developmental Stages

Transient Receptor Potential Melastatin 1(TRPM1)plays an important role in the regulation of fully differentiated melanocyte phenotype and proliferation,and is only expressed in normal melanocytes.It has been shown that the decrease of melanin content is related to the decrease of TRPM1 expression.This experiment was to study the expression characteristics of TRPM1 in different coat color male sheep skin at different developmental stages.We select Dorper(?)× Mongolia sheep(?)0,3,6,12 months black and white hybrid as test objects,extract total RNA of sheep skin tissues and amplify TRPM1 complete CDS The expression characteristics of TRPM1 mRNA in the sheep skin tissues were detected by RT-PCR,immunohistochemical technique was used to locate TRPM1 in sheep skins,Western Blotting technique to analyze TRPM1 protein.The main results were as follows:(1)The complete CDS zone of the sheep TRPM1 gene was 5017 bp and encoded 1,671 amino acids.There are potential signal peptide sites in the secondary structure of TRPM1,containing hydrophobic regions and 6 transmembrane structures,and there may be 9 N-glycosylation sites with potential phosphorylation sites.(2)TRPM1 mRNA has different degree of expression in the black and white hybrid male sheep skin at0?3?6?12 months,expression of 6 months of age white sheep is significantly higher than other months amount of expression in the white sheep skin(P<0.05).The expression of TRPM1 in 0-month-old sheep skin was significantly lower than that of other months(P<0.05).In different development stages of black sheep skin,TRPM1 mRNA expression is significantly higher than expression of the white sheep skin at the same amount of months(P<0.05).(3)Western Blotting results showed that TRPM1 protein was expressed in the skin of black and white sheep at different stages of development,but the expression of different months was different,which further verified the results of mRNA level detection(4)In white and black sheep skin,TRPM1 protein is located in hair follicle hair shaft?outer root sheath?dermal papilla and hair matrixThis study preliminarily illustrates the TRPM1 expression characteristics of different hair color sheep skin at different developmental stages.The preliminary confirmation of the TRPM1 gene plays an important role in the formation of sheep hair color and lays a foundation for the study of the function of sheep TRPM1 gene in the process of melanin formation.