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Identification and Characteristics Analysis of miRNAs with Sexual Differentially Expression at the Early Stages of Sexual Differentiation of Chicken Embryo

Author: HuangPan
Tutor: PengXiuLi；ZuoYanPing
School: Huazhong Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: miRNA Chicken Gonadal development Time expression profiling Spatial expression patterns
CLC: S831
Type: Master's thesis
Year: 2010
Downloads: 112
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Abstract

miRNA, was newly discovered class of post-transcriptional gene expression levels down small molecule non-coding RNA family. Study found that, IniRNA in animal development process plays an important role, but it is in chick embryo gonad differentiation regulation has not been determined. For the study of miRNA in chicken embryos during early sexual differentiation investigate the expression of miRNA in the early development of the role of the gonads, this study pre-chip differentially expressed miRNAs screened for the study, on which the new miRNAs in bioinformatics analysis, and analysis of candidate miRNAs in male and female gonads in the early embryo expression patterns. The results obtained in this study are as follows: 1. Use chicken genomic information to locate the new miRNAs and found that miR-363 and 363 *, miR-551b and miR-612 are located in the chicken chromosome 4 (-), 9 (-) and Z (-), and simultaneously obtained by cloning the miR-363 * in the chicken genome unknown flanking sequences. The secondary structure of the precursor folding results show that the new chicken miRNAs were predicted stem loop precursor structure with mature sequence substantially located in the corresponding stem and folding minimum free energy are ≤ -25kcal/mol. miR-363 (363 *) and miR-551b in human, rat, mouse and chicken four species homology analysis showed that the sequence similarity precursors were 80% or more, and only the mature sequence individual base difference. (2) using RNA-tailing and primer extension and stem loop primer semi-quantitative RT-PCR method to analyze the 13 miRNAs in E3.5-6.5 d female and male gonads time embryo expression profiling, and they are in the same Sex in different periods and in different periods of the same sex differences were statistically significant analysis. The results showed that, overall trend, the detected miRNAs in five were female and 4 showed high expression of male high expression, and the remaining four are irregular expression; miR-363, miR-200b, miR-1810, miR-1599 , miR-551b and miR-196 in the seized certain periods between the sexes differentially expressed achieve significant or very significant level; miR-363, miR-1456, miR-196, miR-363 *, miR-612, miR-1599 , miR-126 and miR-1810 in the same period when the expression of gender exist some significant or very significant. 3 Use WISH technology miR-363 and miR-1810 spatial expression pattern analysis showed that, miR-363 primarily in E4.5 d chicken embryo limbs, ectoderm, notochord and the brain and other parts of the expression in E6.5 d gonadal sites was significantly higher than the kidneys and gonads in females was higher than males; miR-1810 mainly in E4.5 d chicken embryo brain and limbs expressed in E6.5 and E8.5 d gonads expression above the kidneys, and the male gonads was higher than females. Combined results of this study and other species corresponding miRNA studies screened five (miR-363, miR-551b, miR-1810, miR-1599 and miR-200b) may be involved in the process of early embryo gonad miRNAs as the next further object of study.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Poultry > Chicken
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