Identification Of Mitochondrial Genes Affecting The Development Of Oocyte Under FSH Hormone By Using A Microarray Data In Cow

Document Type : Other


Department of Animal Science, University of Khuzestan Agricultural Sciences and Natural Resources, Khuzestan, Iran.


Introduction[1] Oocyte maturity includes nuclear and cytoplasmic maturity, both of which are important for embryo fertilization. Cytoplasmic maturation involves the redistribution of a range of organelles, including mitochondria. The nuclear and cytoplasmic mammalian oocytes maturation is a complex process nuclear maturation is demonstrated by extrusion of first polar body while there may be no indication for cytoplasmic maturation According to critical role of mitochondria for energy production in oocytes, it can be considered as an indicator of cytoplasmic maturation. Oocyte maturation requires more energy. Energy reaches its peak during ovulation. Changes in the mitochondrial distribution pattern can affect the ability of embryo development from oocytes. Since fetal mitochondrial replication is not performed until the blastocyst its stage, mature Oocytes (MII), fertilized Oocytes, Energy required for fertilization, embryonic development prior to implantation and early stages of fetal development depend on the storage of mitochondria in the time of ovulation. Therefore, the location and function of mitochondria can affect the quality of the Oocyte and consequently interfere with the process of embryo development. The topic of genetic networks explores the most important genes in a physiological process. The graph theory is used to construct and reconstruct the biology network. In biology networks, genes, proteins, or any other molecule that plays a role in a cell can be considered as a node and the relationship between these nodes is considered as an edge.
Materials and Methods In this study, GEO access codes for this data set GSE38345 were used to determine the effect of FSH on the expression of mitochondrial genes. In the past decade, with the ability to study genetic information of the genome in a wide range, micro arrays were a high-performance method for analyzing gene expression. The data are microarray and contain the gene expression information for cow's oocyte cells, whose maturity is influenced by the FSH hormone under laboratory conditions. After data implementation, the quality of the data was analyzed and if necessary, normalization was performed using the data conversion technique. Data analysis and comparison of gene expression in two cases before maturation (20 hours after oocyte treatment with FSH in laboratory conditions) and after maturation (96 hours after oocyte treatment with FSH in laboratory conditions) using From the GEO2R software link were done. After identifying the genes and examining the different genes expressed, two genotypes included Increased and decreased expression genes. The interaction of each gene group was studied using a string database based on co-expression data. Gene ontology was performed using the comparative GO database.
Results and Discussion In a comparison between oocyte gene expression data in the pre-maturation stage and the post-maturation stage after treatment with FSH, it was determined that 100 mitochondrial genes in maturation compared to pre-maturation stage increased expression and 94 genes of this organ has declined. Among them, the protein interaction network has been identified in a set of increased and decreased expression genes. Of the 100 genes that have been increased expression, 68 genes are coexpression based on string information. Among decreased expression genes, 53 genes from 64 genes were reported as coexpression. In the protein interaction network of the increased expression genes, the important genes of MRPS10, MRPS18A, MRPL16 and MRPL17, which played a role in the mitochondrial destruction and translation processes of mitochondrial genes, and in the network of decreased expression genes, MRPL22, ATP5B and ATP5C1 genes, which by reducing its expression, attempted to balance in the pathways associated with mitochondrial destruction and ATP production through its role in the ATP synthase structure.
Conclusion The results of this study reveal the most important genes affecting mitochondrial activity during oocyte maturation and control genes of this organ according to the network of protein interactions in the set of increased and decreased expression genes. In addition, the most important biological pathways in order to understand the mechanism of FSH effect on oocyte maturation through mitochondrial organ is investigated. Also, by comprehensive examining the gene expression network in the process of cytoplasmic oocyte maturation and showing the marker genes and different biochemical pathways, it is possible to understand the quality of oocyte during maturation, which can help improve IVM-IVF technique. Since effective mechanisms in cytoplasmic maturity are not yet fully understood, efforts to identify important regulators of mitochondria in oocyte maturation process will be effective in using fertility technology in animal production.


1- Alexeyev, M. F., N. Venediktova, V. Pastukh, I. Shokolenko, G. Bonilla, and G. L. Wilson. 2008. Selective elimination of mutant mitochondrial genomes as therapeutic strategy for the treatment of NARP and MILS syndromes. Gene Therapy, 15(7):516–523.
2- Bader, G. D., M. P. Cary, and C. Sander. 2006. Pathguide: a pathway resource list. Nucleic acids research, 34(2):504–506.
3- Bavister, B. D, and J. M. Squirrell. 2000. Mitochondrial distribution and function in oocytes and early embryos. Human Reporduction. 2:189-198.
4- Behdani, E, and M. R. Bakhtiarizadeh. 2017. Construction of an integrated gene regulatory network link to stress-related immune system in cattle. Genetica, 145(5):441-454.
5- Cantatore, p., Z. Flagella, F. Fracasso, A. M. S. Lezza, M. N. Gadaleta, and A. De Montalvo. 1987. Synthesis and turnover rates of four rat liver mitochondrial RNA species. FEBS Letters, 213(1):144-148.
6- Campoy, E, and M. I. Colombo. 2009. Autophagy in intracellular bacterial infection. Biochimica et Biophysica Acta, 1793(9):1465–1477.
7- Chargui, A. 2012. Subversion of autophagy in adherent invasive Escherichia coli-infected neutrophils induces inflammation and cell death. PLoS One, 7(12):1-10.
8- Chappel, S. 2013. The Role of Mitochondria from Mature Oocyte to Viable Blastocyst. Obstetrics and Gynecology International, 2013(10):1-10.
9- Chen, X., R. Prosser, S. Simonetti, J. Sadlock, G. Jagiello, and E. A. Schon. 1995. Rearranged mitochondrial genomes are present in human oocytes. The American Journal Human Genettics, 57(2):239–247.
10- Endo, T., H. Yamamoto, and M. Esaki. 2003. Functional cooperation and separation of translocators in protein import into mitochondria, the double-membrane bounded organelles. Journal of Cell Science. 116:3259–3267.
11- Feniouk, B. A, and W. Junge. 2005. Regulation of the F0F1-ATP synthase: the conformation of subunit epsilon might be determined by directionality of subunit gamma rotation .FEBS Letters, 579(23):5114-5118.
12- Ferreira, E.M., A. A. Vireque, P. R. Adona, F. V. Meirelles, R. A. Ferriani, and P. A .Navarro. 2009. Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence. Theriogenology, 71(5):836-848.
13- Fleckenstein, B., M. D. Daniel, R. D. Hunt, R. D. Werner, L. A. Falk, and C. Mulder. 1978. Tumour induction with DNA of oncogenic primate herpesviruses. Nature. 274:57–59.
14- Ghanem, N., M. Holker, F. Rings, D. Jennen, E. Tholen, M. A. Sirard, H. Torner, W. Kanitz, K. Schellander, and D. Tesfaye. 2007. Alterations in transcript abundance of bovine oocytes recovered at growth and dominance phases of the first follicular wave. BMC Developmental Biologhy. 7:90.
15- Gomes, L. C, and I. Dikic. 2014. Autophagy in antimicrobial immunity. Molecular Cell, 54(2):224–233.
16- Israel, M. A., H. W. Chan, S. L. Hourihan, W. P. Rowe, and M. A. Martin. 1979. Biological activity of polyomavirus DNA in mice and hamsters. Journal Virologhy. 29:990–996.
17- Jansen, R. P, and K. de Boer .1998. The bottleneck: mitochondrial imperatives in oogenesis and ovarian follicular fate. Molecular and Cellular Endocrinologhy, 145(1):81-88.
18- Kenmochi, N., T. Suzuki, T. Uechi, M. Magoori, M. Kuniba, S. Higa, K. Watanabe, and T. Tanaka. 2001. The human mitochondrial ribosomal protein genes: mapping of 54 genes to the chromosomes and implications for human disorders. Genomics, 77(2):65-70.
19- Krisher, R. L. 2004. The effect of oocyte quality on development. Journal of Animal Science. 82:14-23.
20- Labrecque, R. M., C .Vigneault, P .Blondin, and M. A. Sirard. 2013. Gene Expression Analysis of Bovine Oocytes With High Developmental Competence Obtained From FSH-Stimulated Animals. Molecular Reproduction & Development, 80(6):428–440.
21- Lapaquette, P., P. Brest, P. Hofman, and A. Darfeuille-Michaud . 2012. Etiology of Crohn’s disease: many roads lead to autophagy. Journal of Molecoular Medicine, 90(9):987–996.
22- Liu, J., L. Jing, and X. TU. 2016. Weighted gene co-expression network analysis identifies specific modules and hub genes related to coronary artery disease. BMC Cardiovasc Disord, 5(16):54.
23- Lu, S., J. Arthos, D. C. Montefiori, Y. Yasutomi, K. Manson, F. Mustafa, E. Johnson, J. C. Santoro, J. Wissink, J. I. Mullins, J. R. Haynes, N. L. Letvin, M. Wyand, and H. L. Robinson. 1996. Simian immune deficiency virus DNA vaccine trial in macaques. Virology journal, 70(6):3978–3991.
24- Luo, X., I. Budihardjo, H. Zou, C. Slaughter, and X. Wang .1998. Bid, a BCL2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell, 94(4):481-490.
25- Nagai, S., T. Mabuchi, S. Hirata, T. Shoda, T. Kasai, S. Yokota, H. Shitara, H. Yonekawa, and K. Hoshi. 2006. Correlation of abnormal mitochondrial distribution in mouse oocytes with reduced developmental competence. The Tohoku Journal Experimental Medicine, 210(2):137-144.
26- Nichols, J., B. Zevnik, K. Anastassiadis, H. Niwa, D. Klewe-Nebenius, I. Chambers, H. Schöler, and A. Smith. 1998. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell, 95(3):379-391.
27- Nicholls, D. G., S. J. Ferguson, and S. Ferguson. 2003. Bioenergetics. Third Edition.
28- Nishi, Y., T. Takeshita, K. Sato, and T. Araki. 2003. Change of the mitochondrial distribution in mouse ooplasm during in vitro maturation. Journal of Nippon Medical School, 70(5):408-415.
29- Nivet, A. L., A. Bunel, R. Labrecque, J. Belanger, C. Vigneault, P. Blondin, and M. A. Sirard. 2012. FSH with drawal improves developmental competence of oocytes in the bovine model. Reproduction, 143(2):165-171.
30- Novak, I. 2012. Mitophagy: a complex mechanism of mitochondrial removal. Antioxidants & Redox Signaling, 17(5):794–802.
31- Ohta, S., H. Tomura, K. Matsuda, and Y. Kagawa. 1988. Gene structure of the human mitochondrial adenosine triphosphate synthase beta subunit. The Journal Of Biological Chemistry, 263(23):11257–11262.
32- Patel, O. V., A. Bettegowda, J. J. Ireland, P. M. Coussens, P. Lonergan, and G. W. Smith. 2007. Functional genomics studies of oocyte competence: Evidence that reduced transcript abundance for follistatin is associated with poor developmental competence of bovine oocytes. Reproduction, 133(1):95-106.
33- Peddinti, D., E. Memili, and S. C. Burgess. .2010. Proteomics-Based Systems Biology Modeling of Bovine Germinal Vesicle Stage Oocyte and Cumulus Cell Interaction. PLoS One, 5(6):1-13.
34- Riquelme Medina, I, and Z .Lubovac-Pilav .2016. Gene Co-Expression Network Analysis for Identifying Modules and Functionally Enriched Pathways in Type 1 Diabetes. PLoS One, 11(6):1-18.
35- Segal, E., N. Friedman, N. Kaminski, A. Regev, and D. Koller. 2005. From signatures to models: understanding cancer using microarrays. Nature Genetics, 37 (l):38-45.
36- Shuster, R. C., A. J. Rubenstein, and D. C. Wallace. 1988. Mitochondrial DNA in anucleate human blood cells. Biochemical and Biophysical Research Communications, 155(3):1360-1365.
37- Springer, M. Z, and K. F. Macleod .2016. Mitophagy: mechanisms and role in human disease. The Journal of Pathology, 240(3):253–255.
38- Stojkovic, M., S. A. Machado, P. Stojkovic, V. Zakhartchenko, P. Hutzler, P. B. Gonçalves, and E. Wolf. 2001. Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and after in vitro maturation: correlation with morphological criteria and developmental capacity after in vitro fertilization and culture. Biology of Reproduction, 64(3):904–909.
39- Susin, S. A., N. Zamzami, M. Castedo, T. Hirsch, P. Marchetti, A. Macho, E. Daugas, M. Geuskens, and G. Kroemer. 1996. BCL-2 inhibits the mitochondrial release of an apoptogenic protease. Journal. Experimental. Medicine, 184(4):1331-1341.
40- Uddin, R. K, and S. M Singh. 2013. Hippocampal gene expression meta-analysis identifies aging and age-associated spatial learning impairment (ASLI) genes and pathways. PLoS One, 8(7):1-15.
41- Van Blerkom, J .1991. Microtubule mediation of cytoplasmic and nuclear maturation during the early stages of resumed meiosis in cultured mouse oocytes. Proceedings National Academy of Sciences USA, 88(11):5031–5035.
42- Van Blerkom, J. 2004. “Mitochondria in human oogenesis and preimplantation embryogenesis: engines of metabolism, ionic regulation and developmental competence. Reproduction, 128(3):269–280.
43- Wallace, D. C. 2001. A mitochondrial paradigm for degenerative diseases and ageing. Novartis Foundation Symposia. 235:247-263.
44- Wilding, M., B. Dale, M. Marino, L. di Matteo, C. Alviggi, M. L. Pisaturo, L. Lombardi, and G. De Placido. 2001. Mitochondrial aggregation patterns and activity in human oocytes and preimplantation embryos. Human Reproduction, 16(5):909-917.
45- Yuk, J. M., T .Yoshimori, and E. K. Jo. 2012. Autophagy and bacterial infectious diseases. Experimental and Molecular Medicine, 44(2):99-108.
46- Zhang, X., X. Q. Wu, S. Lu, Y. L. Guo, and X. Ma. 2006. Deficit of mitochondria-derived ATP during oxidative stress impairs mouse MII oocyte spindles. Cell Research, 16(10):841–850.
  • Receive Date: 01 January 1970
  • Revise Date: 03 July 2019
  • Accept Date: 24 August 2019
  • First Publish Date: 21 June 2020