The Research Of Albino Trait Mechanism Of Cricetulus Barabensis Albino(A：CHA)

Cultivation of Cricetulus barabensis albino mutant lines (A: CHA) at home and abroad areconsidered for the first time, The strains of Medical Biology provides a new animal model, alsohas a high academic value for the field of Laboratory Animal Care International that adds a newstrain. Cricetulus barabensis albino mutant lines (A: CHA) after establishment shows betterapplication prospect than the black line hamster in the skin microcirculation, radiation protection,ectoparasites and epidemiological applications, which have more superiority than the black linehamster. Currently only know the black line hamster albino mutant lines genes that belong to asingle genetic recessive inheritance, but do not know the reason of black line hamster in albinotraits, so it is necessary to reveal the mechanism of black line hamsters in albino mutation ofalbino trait, figure out the mutations in the genetic background of the population, to furtherimprove the albino black line hamster mutant group on the basic of biological data, to lay a goodfoundation for the popularization and application in biomedical. This study use the skin tissue ofblack line hamster albino mutation as research material, using the methods of biology, histology,molecular biology to comparative study the skin tissue of black line hamster albino mutant line,comprehensively discussing the machanism of the albino characteristics of the Cricetulusbarabensis albino line. The thesis includes two parts, the first literature of thesis is review, thesecond literature of thesis is research papers, including five chapters, and the last is the summaryand outlook. The main research content includes histological analysis of the melanocytes withinhair folliciles which locate in black line hamster and albino mutant lines, comparing skin tissuemelanoma cell ultrastructure, albino gene cloning and bioinformatics analysis, albino relatedgenes expression and analysis of the albino correlation, based on Illumina sequencing technologyto transcriptome sequencing and analysis of the skin tissue which is in black line hamster andalbino mutant lines. Experiment results will now report as follows:1. Histological analysis of the hair follicle melanocytes which locate inCricetulus barabensis and its albino mutant linesDopa staining results showed that Cricetulus barabensis and its albino mutation of skin tissuewere positive staining of dopamine, dobutamine positive staining shows the dopa positive melanin cells, this results demonstrate that mature melanocytes within black line hamsters and albinomutant lines are distributed in the skin tissue, but the distribution is different. Black line hamsterin dopa-positive area was significantly higher than the albino mutant lines. Dopa is a specificsubstrate of tyrosinase, shows that the TYR expression level of albino mutation was significantlylower than the black line hamster. Dopa-toluidine blue redyeing fully indicates that thedistribution of dopa-positive melanin in the skin tissue, the skin tissue melanocytes which locatein the black line and albino hamster exist in the hair follicle dermal papilla.2. Cricetulus barabensis and albino mutant skin melanoma cellultrastructure study(1) Cricetulus barabensis melanin content was significantly higher than the albino mutant.Visible, lack of melanin in the skin tissue is the direct reason that causing Cricetulus barabensisalbino mutation.(2) Comparing with Cricetulus barabensis, the albino mutation has significantly fewer skinmelanin cells, the development of melanosomes in melanocytes is low, most melanosomes arepre-melanosome that has not deposit of melanin. For the albino mutation, the density ofmelanosomes in melanocytes is obviously smaller. Visible, the lack of melanocytes and lack ofmelanin deposition in melanosomes are the histologic features of Cricetulus barabensis albinooccurrence.3. Cloning and sequence analysis about an albino genes that is in Cricetulusbarabensis and its albino mutationIn this test, through the Cricetulus barabensis and albino mutation of TYR, TYRP1, TYRP2,P, MITF, Agouti gene cloning and sequence analysis, clear Cricetulus barabensis albino mutantalbino trait is due to the gene mutation, according to the site of gene conservation in mice and ratsdesign a few pairs of primers, using RT-PCR method to clone and sequence. Successfully obtainthe Cricetulus barabensis and albino mutation of TYR, TYRP1, TYRP2, P, MITF, agouti gene,first clone coding region sequence of TYR, TYRP1, TYRP2, P, MITF, Agouti gene which is inCricetulus barabensis and albino mutation, but from coding regions did not mutant, the albino traitmechanism needs further research.4. Comparative analysis about the level of an albino genes expression which isin Cricetulus barabensis and albino mutationThe results showed that TYR, TYRP1, TYRP2, P, MITF, Agouti mRNA expression quantityin black line hamsters is2.5、5.3、1.12、1.7、1.8、1.7time to albino mutation, the conclusion prompt that the expression level of TYR, TYRP1, TYRP2, P, MITF, Agouti has certain correlationwith Cricetulus barabensis color phenotype, suggesting that several members play a regulatingrole in the pigmentation process. This study clarify an albino genes regulation mechanism ofmelanin traits, as well as laid a theoretical foundation of the mechanism for Cricetulus barabensisalbino mutation albino traits.5. Based on Illumina sequencing technology to transcriptome sequencingand analysis of the skin tissue which is in Cricetulus barabensis and albinomutant linesThis study take the skin tissue of black line hamsters and albino mutation as the researchobject, using Illumina/Solexa high-throughput sequencing technology to the depth of theirtranscriptome sequencing, using short sequence assembly software SOAP denovo to de novoassembly the results of its transcriptome sequencing, establishing database of the transcriptome,and proceeding sequence alignment, functional annotation and metabolic pathway analysis forprotein database with nr, Swiss-Prot, KEGG and COG; on this basis, proceeding the analysis ofdifferentially expressed genes, screening pigment synthesis genes, and to explore the bleachingmechanism.Through this study, a total of more than80million high quality matching short sequencesare obtained, through sequence assembly received116,504Unigenes. By black line hamsters andalbino mutation skin transcriptome analysis of differentially expressed genes were screened16,905differentially expressed genes(|log2Ratio︱≥1and FDR≤0.001), through GO functionenrichment analysis and KEGG pathway enrichment analysis, these differentially expressed genesinvolved in cellular metabolism, protein translation, extracellular matrix receptor interaction andbiological processes. These differentially expressed genes were screened out53kinds of geneswhich play important regulating effect in protein synthesis, cell proliferation and cell apoptosisduring the process of growth in black line hamsters,12kinds of differentially expressed geneswhich is related to the growth of black line hamsters, and7kinds of differentially expressed geneswhich is ralated to pigment biosynthesis, the results of these experiments lay the foundation forfurther study about the structure and function of genes which is in black line hamsters.