Dissertation > Excellent graduate degree dissertation topics show

Expression of β-Catenin in Pig’s Ovary and the Effect of β-Catenin on Porcine Granulosa Cells Apotosis and Steroidogenesis Related Enzyme

Author: WangLi
Tutor: XuYinXue
School: Nanjing Agricultural College
Course: Animal Genetic Breeding and Reproduction
Keywords: pig granulosa cell Wnt/β-catenin signaling pathway RNAi over-expression apoptosis steroid hormone synthesis
CLC: S828
Type: Master's thesis
Year: 2010
Downloads: 15
Quote: 0
Read: Download Dissertation

Abstract


P-catenin is a critical signal transduction molecules of the Wnt signaling pathway plays an vital role in the female during development, deletion or mutation ofβ-catenin will lead to Wnt/β-catenin signaling pathway disorder, and thus have impact on body tissues or cells. Wnt4 knockout mice before birth can lead to some of the sex reversal and the prostration of oocytes, P-catenin knockout mouse develop follicular containing pleomorphic follicular granulosa cells, which lead to the occurrence of granular cell carcinoma. Recently found thatβ-catenin involved in transcriptional regulation network of gonadotropin-releasing hormone, plays an important role in hormone formation.In this study, Erhualian pig, Pietrain Pig, commercial pigs and pig ovarian granulosa cells were taken as experimental material, real time RT-PCR were utilized to detect the different expression ofβ-catenin within the pig ovary and ovarian tissue;β-catenin was knocked down by RNAi methods, and based on the principle that LiCl can antagonize the GSK-3βto phosphorylate of P-catenin thus degradation, which will lead to the accumulation of P-catenin in nucleus.granulosa cells were continuously treated with LiCl to over-express P-catenin in GCs. Flow cytometry, real time PCR, western blot and other methods were used to examine the effect of P-catenin on granulosa cell apoptosis and steroid hormone synthesis from silencing and over-expressing ofβ-catenin gene in pig ovarian granulosa cell.The main results are as follows:1 Real time PCR was explored to detect the expression ofβ-catenin, the key signal transduction molecule of Wnt/β-catenin signaling pathway,in different ovary tissues and in ovaries of different varieties of porcine.The results show that:β-catenin in porcine of different fertility differentially expressed in ovary, the expression ofβ-catenin in erhualian pig is over 3 times of that in commercial pig ovary. While the expression ofβ-catenin in Pietrain pig is of 40% of that in commercial pig ovary.β-catenin is differently expressed in different development stage,the expression ofβ-catenin in large follicles was significantly higher than in the medium follicles and small follicles (p<0.05).β-catenin in ovarian granulosa cells was also is nearly 2.5 times of that in oocytes(p<0.05).2 To demonstrate the effects of Wnt/β-catenin signal on cell apoptosis and steroid hormone synthesis of porcine GCs, granulosa cells were isolated from 3-5mm antral follicles in diameter and cultured in vitro. Based onβ-catenin in the core molecular of the pathway, A strategy of RNAi-mediated’gene silencing’ofβ-catenin were used to "silence" pigβ-catenin gene, inhibit Wnt/β-catenin signaling pathway, while LiCl was used to activate the Wnt/β-catenin signaling pathway, to detect the effect ofβ-catenin on porcine granulosa cell apoptosis and steroid hormone synthesis.The results showed that:(1) P-catenin gene successfully "knocked down" by using the RNAi technology, mRNA levels ofβ-catenin was reduced by 59.93%(p<0.05), protein levels were also decreased. Knocked down P-catenin significantly increased granulosa cell apoptosis rate, by up-regulating Bax by 32.76%(p<0.05)and down-regulated BCL-2 by 44.28%(p<0.05), the expression of P450arom mRNA was reduced by 47.72%(p<0.05), but the expression of P450scc was not affected. (2) LiC1 treatment successfully activated Wnt/β-catenin pathway,β-catenin mRNA levels was up-regulated by 95.15%(p<0.05), protein levels was increased. Over-expression ofβ-catenin was significantly increased apoptosis rate,by down-regulating Bax mRNA by 25.60%(p<0.05) and increasing BCL-2 mRNA by 12.58%(p>0.05), and P450arom mRNA was up-regulated by 151.6%(p<0.05), P450scc mRNA was upregulated by 124.3%(p<0.05). Indicate that the Wnt/β-catenin signaling pathway have important role in porcine granulosa cells.

Related Dissertations

  1. Research on Combinatorial Regulation of Multiple Transcription Factors,Q78
  2. The Research of the High-Rise Building’s Form and Expression in the New Century,TU971
  3. Research on Design of Teaching Buildings for Arts Departments of Higher Learning Institutions,TU244.3
  4. Research on Temporal Information Recognition and Normalization,TP391.1
  5. Expression of D-AtCGS in E. Coli and Preparation of Polyclonal Antibody Against D-AtCGS,Q943.2
  6. The Study on Arabidopsis Thaliana Heat Shock Factor HSFA 1d Response to Formaldehyde Stress,Q945.78
  7. Expression of hBMP4 and hBMP7 in Chinese Hamster Ovary Cells,Q78
  8. Cloning and Expression Analysis of GPx, GST and SAHH Genes in Chlamydomonas Sp. ICE-L from Antarctica,Q943.2
  9. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  10. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  11. Expression of Prolyl Isomerase Pin1 in Osteosarcoma and the Effect of Regulation on Cell Cycle,R738.1
  12. The Expression Survey of BMP Signalling Pathway in the Human Embryonic Tooth Germ,R78
  13. Impact on Drosophila Brain Developement Through the Overexpression and Loss of Function of Gsα,Q75
  14. The Study on Cryopreservation and Mechanism of Freezing Injury on the Spermatozoa of Coelomactra Antiquate,S968.3
  15. Preliminary Study of Phosphorylation Effect on UGT1A3 Metabolism Activity Towards Drugs,R346
  16. The Anti-tumor Effect of CADPE and Inducing Apoptosis in Human Gastric Cancer Cells,R735.2
  17. Cloth and β- elemene combined administration of anti-tumor effect and mechanism of celecoxib,R96
  18. Cloning and Expression of Growth Hormone Gene in Cobia (Rachycentron Canadus),Q786
  19. Cloning and Characterization of BAFF Gene from Whitespotted Catshark and Crucian Crap,S917.4
  20. Improvement of the Resistance of Nicotiana Tabaccum to Virus by RNAi and Initial Establishment of Genetic Transformation System of L.regale,S435.72
  21. TRAIL in the regulation of tumor invasion CD4 ~ CD25 ~ Treg,R730.2

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Pig
© 2012 www.DissertationTopic.Net  Mobile