Expression And Location Analysis Of APC In Alpaca Skin

Mammal coat color is decided by pigment kinds, numbers in melanocytes. Researches show that Wnt signal pathway involves in the forming process of coat color, and regulatory factors in this path which are play impotant roles in melanocytes development include Wnt、 APC、(3-catenin、LEF and MITF. In recent years, reseachers conducted mass study on the interaction of these regulatory factors, but concentrate mainly on cancer mechanism parts. And, our lab studied Wnt3a、β-catenin、LEF and MITF respectively, but no study about expression and location of APC in animal skin. In this reseach, we set alpaca which has variety coat color phenotypic as the object of study, using quantitative real-time PCR and immunohistochemistry to detect the mRNA and protein expression levels of APC in brown and white coat color alpaca, to explore the mechanism of APC action in forming process of alpaca coat color.1Logining NCBI, look up APC mRNA sequences in cow and other species, find conservative region, using Primer3.0plus soft to design the primer of APC. Using PCR, and amplified a248bp length of gene sequence, after sequencing and the Blast function of NCBI, detected that this sequence has a high homology with other species, so ensured that it is APC gene sequence.2Using using quantitative real-time PCR to analyse the mRNA expression level of APC in brown and white coat color alpaca, the result showes that:The gene expression quantity of APC in brown alpaca skin was5.0422times than white one, and hints that APC may involve in the coat color formation in alpaca.3Using immunohistochemistry to analyse the location of APC in alpaca skin, the result showes that APC exprese in hair bulb and root of hair follicles and epidermis, and the average protein expression of APC in brown alpaca skin was higher than that in white one by using the analysis of Image-Pro Plus6.0..Thus, APC express in alpaca skin, and whether in mRNA and in protein level, browns are higher than white ones, the results show that APC may involve in the coat color formation in alpaca, and we speculate that APC may up-regulate β-cantenin, and prompt the function of LEF and MITF, then regulate the coat color.