Investigation of stem cells in adult and prepubertal mouse ovaries


The possible presence of oocyte and granulosa cells originated from stem cells in the adult mammalian ovaries was claimed by some studies which will lead to major changes in reproductive biology and infertility treatments. Purpose of this research is to investigate the possible existence and the location of the potential stem cells in mouse ovaries. In this study, the ovaries from 2-week (pre-puberty) and 8-week (adult) old BALB-C mice were used. For the investigation of the presence of possible stem cells, the expression profiles of three well known stem cell markers, Oct-4, Nanog and Sox2 were determined in the ovaries of two different age groups by real time quantitative RT-PCR (qRT-PCR). Protein expression levels and their localization in the ovary cells were immunohistochemically evaluated on fresh-frozen ovary tissue sections by using monoclonal antibodies specific to Sox2, Nanog and Oct-4. The gene expression levels of Oct-4 and Nanog were found to be significantly differentiated between 2-week old and 8-week old mice whereas no significant difference was observed in the expression level of Sox2 between two age groups. Immunohistochemistry results showed the presence of both Sox2 and Oct-4 protein in the cytoplasm of ovarian epithelial cells, granulosa cells, oocytes and theca cells. Nanog protein was observed only in the nucleus of the oocytes and furthermore the expression of Nanog was higher in eight weeks old samples compared to two weeks old ones according to qRT-PCR results. These results suggest for the first time that Nanog protein is expressed both in adult and pre-puberty mouse ovaries and locate at the nucleus of the oocytes and to the best of our knowledge this is the first study that shows the differential expression of Oct-4, Nanog and Sox2 in pre-puberty and adult mouse ovaries by qRT-PCR. Collectively, our results may suggest that both pre-puberty and adult mice ovaries accommodate cells carrying stem cell features.

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Esmaeilian, Y. , Dedeoglu, B. , Atalay, A. and Erdemli, E. (2012) Investigation of stem cells in adult and prepubertal mouse ovaries. Advances in Bioscience and Biotechnology, 3, 936-944. doi: 10.4236/abb.2012.37115.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Johnson, J., Canning, J., Kaneko, T., Pru, J.K. and Tilly, J.L. (2004) Germline stem cells and follicular renewal in the postnatal mammalian ovary. Nature, 428, 145-150. doi:10.1038/nature02316
[2] Bukovsky, A., Svetlikova, M. and Caudle, M.R. (2005) Oogenesis in cultures derived from adult human ovaries. Reproductive Biology and Endocrinology, 3, 17. doi:10.1186/1477-7827-3-17
[3] Virant-Klun, I., Zech, N., Rozman, P., Vogler, A., Cvjeticanin, B., Klemenc, P., Malicev, E. and Meden-Vrtovec, H. (2008) Putative stem cells with an embryonic character isolated from the ovarian surface epithelium of women with no naturally present follicles and oocytes. Differentiation, 76, 843-856. doi:10.1111/j.1432-0436.2008.00268.x
[4] Honda, A., Hirose, M., Hara, K., Matoba, S., Inoue, K., Miki, H., Hiura, H., Kanatsu-Shinohara, M., Kanai, Y. and Kono, T. (2007) Isolation, characterization, and in vitro and in vivo differentiation of putative thecal stem cells. Proceedings of the National Academy of Sciences USA, 104, 12389-12394. doi:10.1073/pnas.0703787104
[5] Liu, Y., Wu, C., Lyu, Q., Yang, D., Albertini, D.F., Keefe, D.L. and Liu, L. (2007) Germline stem cells and neooogenesis in the adult human ovary. Developmental Biology, 306, 112-120. doi:10.1016/j.ydbio.2007.03.006
[6] Boyer, L.A., Mathur, D. and Jaenisch, R. (2006) Molecular control of pluripotency. Current Opinion in Genetics & Development, 16, 455-462. doi:10.1016/j.gde.2006.08.009
[7] Chambers, I., Colby, D., Robertson, M., Nichols, J., Lee, S., Tweedie, S. and Smith, A.G. (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell, 113, 643-655. doi:10.1016/S0092-8674(03)00392-1
[8] Mitsui, K., Tokuzawa, Y., Itoh, H., Segawa, K., Murakami, M., Takahashi, K., Maruyama, M., Maeda, M. and Yamanaka, S. (2003) The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell, 113, 631-642. doi:10.1016/S0092-8674(03)00393-3
[9] Boyer, L.A., Lee, T.I., Cole, M.F., Johnstone, S.E., Levine, S.S., Zucker, J.P., Guenther, M.G., Kumar, R.M., Murray, H.L., Jenner, R.G., Gifford, D.K., Melton, D.A., Jaenisch, R. and Young R.A. (2005) Core transcriptional regulatory circuitry in human embryonic stem cells. Cell, 122, 947-956. doi:10.1016/j.cell.2005.08.020
[10] Rodda, D.J., Chew, J., Lim, L.H., Loh, Y.H., Wang, B., Ng, H.H. and Robson, P. (2005) Transcriptional Regulation of Nanog by OCT4 and SOX2. Journal of Biological Chemistry, 280, 24731-24737. doi:10.1074/jbc.M502573200
[11] Masui, S., Nakatake, Y., Toyooka, Y., Shimosato, D., Yagi, R., Takahashi, K., Okochi, H., Okuda, A., Matoba, R., Sharov, A.A., Ko, M.S. and Niwa, H. (2007) Pluripotency governed by Sox2 via regulation of Oct 3/4 expression in mouse embryonic stem cells. Nature Cell Biology, 9, 625-635. doi:10.1038/ncb1589
[12] Zou, K., Yuan, Z., Yang, Z., Luo, H., Sun, K., Zhou, L., Xiang, J., Shi, L., Yu, Q., Zhang, Y., Hou, R. and Wu, J. (2009) Production of offspring from a germline stem cell line derived from neonatal ovaries. Nature Cell Biology, 11, 631-636. doi:10.1038/ncb1869
[13] Cauffman, G., Van de Velde, H., Liebaers, I. and Van Steirteghem, A. (2005) Oct-4 mRNA and protein expression during human preimplantation development. Molecular Human Reproduction, 11, 173-181. doi:10.1093/molehr/gah155
[14] Zuccotti, M., Merico, V., Sacchi, L., Bellone, M., Brink, T.C., Stefanelli, M., Redi, C.A., Bellazzi, R., Adjaye, J. and Garagna, S. (2009) Oct-4 regulates the expression of Stella and Foxj2 at the Nanog locus: implications for the developmental competence of mouse oocytes. Human Reproduction, 24, 2225-2237. doi:10.1093/humrep/dep191
[15] Avilion, A.A., Nicolis, S.K., Pevny, L.H., Perez, L., Vivian, N. and Lovell-Badge, R. (2003) Multipotent cell lineages in early mouse development depend on SOX2 function. Genes & Development, 17, 126-140. doi:10.1101/gad.224503
[16] Yamaguchi, S., Kimura, H., Tada, M., Nakatsuji, N. and Tada, T. (2005) Nanog expression in mouse germ cell development. Gene Expression Patterns, 5, 639-646. doi:10.1016/j.modgep.2005.03.001
[17] Hart, A.H., Hartley, L., Ibrahim, M. and Robb, L. (2004) Identification, cloning and expression analysis of the pluripotency promoting Nanog genes in mouse and human. Developmental Dynamics, 230, 187-198. doi:10.1002/dvdy.20034
[18] Niwa, H., Miyazaki, J. and Smith, A.G. (2000) Quantitative expression of Oct 3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nature Genetics, 24, 372-276. doi:10.1038/74199
[19] Whiteside, S.T. and Goodbourn, S. (1993) Signal transduction and nuclear targeting: regulation of transcription factor activity by subcellular localisation. Journal of Cell Science, 104, 949-955.
[20] Bukovsky, A., Caudle, M.R., Virant-Klun, I., Gupta, S.K., Dominguez, R., Svetlikova, M. and Xu, F. (2009) Immune physiology and oogenesis in fetal and adult humans, ovarian infertility, and totipotency of adult ovarian stem cells. Birth Defects Research Part C, 87, 64-89.

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