| Antlers are the only boneness organ that can be completely regenerated in mammals,which is a very precious animal tissue regeneration model in nature.In the fast growing period,although the cell proliferations speed of the antler growth center is more than 30 times that of the cancer cell proliferation,there is no sign of canceration in the antler cells.Therefore,researching the antler regeneration mechanism and the role growth factor played in the antler cell proliferation process can provide new ideas and directions for human cancer treatment.Reindeer are the only deer species in both female and male horned,the phenomenon denies that horn is the second sexual characteristic of the male deer.However,the female reindeer horn has obvious differences with the male reindeer,such as antler shape,number of branches and regrowth time.Therefore,there is great scientific significance to research the differences in the expression of antler growth-related genes in male and female reindeer antler for understanding the role of various growth factors in the growth process and the molecular regulatory mechanisms of reindeer antler.In this study,the velvet skin tissue of reindeer antler-tip was used as the research material,and antler growth-related TGF??EGFR?Morf411?MRFAP1?IGF1?.PTN genes were selected as the research object.Trizol method was used to isloate total RNA from the velvet skin tissues of the reindeer antler-tip,and the cDNA sequences of the growth-related genes of the reindeer antler were obtained by RT-PCR and molecular cloning techniques,then bioinformatics methods were used to depthly analysis of the biological information carried by each gene.Finally,primers were designed based on the known cDNA sequences of gene,and real-time fluorescent quantitative PCR was used to study the differential expression of antler growth related genes in the velvet skin tissue of male and female reindeer antler.The results were as follows:(1)The sequences of TGFa,EGFR,Morf411,MRFAP1,IGF1,PTN genes of antler growth-related of reindeer were cloned and obtained,which sizes are 499 bp,3529 bp,1097 bp,532 bp,680 bp,and 787 bp.After analysis,it was found that the cDNA of TGFa,Morf411,MRFAP1,IGF 1,and PTN genes contained the complete coding region sequences,and EGFR contained partial coding region sequences,based on the cDNA sequence analysis,a large number of biological information of antler growth-related genes were found.(2)Through multiple sequence comparisons and phylogenetic tree analysis,we found that antler growth-related genes of reindeer were very conservative in evolution,and the similarities of the corresponding genes cDNA sequences of homologous species is all above 95%,such as cattle and sheep.The amino acid sequences of the encoded protein could reach 97%or higher similarity with corresponding proteins of homologous species,such as cattle and sheep,and there were fewer differences among species.Therefore,it is presumed to be a key functional gene in the evolution of the species and may participate in the regulation of cell proliferation in the reindeer antler during the rapid growth period.(3)Fluorescent quantitative analysis revealed that there was a difference in the expression of antler growth-related genes between velvet skin of the male and female reindeer antlers.The expression levels of TGFa,EGFR,Morf411 and MRFAP1 genes in the antler velvet skin tissue of female reindeer were 3.377±0.226,3.289±0.272,2.127±0.032,and 1.992±0.058 times higher than that of the male,The expression levels of IGF 1 gene expression levels in the antler velvet skin tissue of male reindeer was 3.725±0.060 times higher than that of the female,The expression levels of PTN gene expression levels in the antler velvet skin tissue of female reindeer was 1.271 ±0.034 times higher than that of the male.Compared with other genes,the expression levels of PTN gene showed less difference in the antler velvet skin tissue between the male and female reindeer. |
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