Dynamic Changes in EPCAM Expression During Spermatogonial Stem Cell Differentiation in the Mouse Testis

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2011 Aug 23

PMID 21858196

Citations 21

Authors

Mito Kanatsu-Shinohara

Seiji Takashima

Kei Ishii

Takashi Shinohara

Affiliations

Soon will be listed here.

Abstract

Background: Spermatogonial stem cells (SSCs) have the unique ability to undergo self-renewal division. However, these cells are morphologically indistinguishable from committed spermatogonia, which have limited mitotic activity. To establish a system for SSC purification, we analyzed the expression of SSC markers CD9 and epithelial cell adhesion molecule (EPCAM), both of which are also expressed on embryonic stem (ES) cells. We examined the correlation between their expression patterns and SSC activities.

Methodology And Principal Findings: By magnetic cell sorting, we found that EPCAM-selected mouse germ cells have limited clonogenic potential in vitro. Moreover, these cells showed stronger expression of progenitor markers than CD9-selected cells, which are significantly more enriched in SSCs. Fluorescence-activated cell sorting of CD9-selected cells indicated a significantly higher frequency of SSCs among the CD9(+)EPCAM(low/-) population than among the CD9(+)EPCAM(+) population. Overexpression of the active form of EPCAM in germline stem (GS) cell cultures did not significantly influence SSC activity, whereas EPCAM suppression by short hairpin RNA compromised GS cell proliferation and increased the concentration of SSCs, as revealed by germ cell transplantation.

Conclusions/significance: These results show that SSCs are the most concentrated in CD9(+)EPCAM(low/-) population and also suggest that EPCAM plays an important role in progenitor cell amplification in the mouse spermatogenic system. The establishment of a method to distinguish progenitor spermatogonia from SSCs will be useful for developing an improved purification strategy for SSCs from testis cells.

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References

Kubota H, Avarbock M, Brinster R . Spermatogonial stem cells share some, but not all, phenotypic and functional characteristics with other stem cells. Proc Natl Acad Sci U S A. 2003; 100(11):6487-92. PMC: 164473. DOI: 10.1073/pnas.0631767100. View

Kanatsu-Shinohara M, Ogonuki N, Inoue K, Miki H, Ogura A, Toyokuni S . Long-term proliferation in culture and germline transmission of mouse male germline stem cells. Biol Reprod. 2003; 69(2):612-6. DOI: 10.1095/biolreprod.103.017012. View

Oatley J, Brinster R . Regulation of spermatogonial stem cell self-renewal in mammals. Annu Rev Cell Dev Biol. 2008; 24:263-86. PMC: 4066667. DOI: 10.1146/annurev.cellbio.24.110707.175355. View

Shinohara T, Avarbock M, Brinster R . Functional analysis of spermatogonial stem cells in Steel and cryptorchid infertile mouse models. Dev Biol. 2001; 220(2):401-11. DOI: 10.1006/dbio.2000.9655. View

Lolicato F, Marino R, Paronetto M, Pellegrini M, Dolci S, Geremia R . Potential role of Nanos3 in maintaining the undifferentiated spermatogonia population. Dev Biol. 2007; 313(2):725-38. DOI: 10.1016/j.ydbio.2007.11.011. View

Munz M, Baeuerle P, Gires O . The emerging role of EpCAM in cancer and stem cell signaling. Cancer Res. 2009; 69(14):5627-9. DOI: 10.1158/0008-5472.CAN-09-0654. View

Morimoto H, Kanatsu-Shinohara M, Takashima S, Chuma S, Nakatsuji N, Takehashi M . Phenotypic plasticity of mouse spermatogonial stem cells. PLoS One. 2009; 4(11):e7909. PMC: 2774941. DOI: 10.1371/journal.pone.0007909. View

Tegelenbosch R, de Rooij D . A quantitative study of spermatogonial multiplication and stem cell renewal in the C3H/101 F1 hybrid mouse. Mutat Res. 1993; 290(2):193-200. DOI: 10.1016/0027-5107(93)90159-d. View

Lu T, Lu R, Liao M, Yu J, Chung C, Kao C . Epithelial cell adhesion molecule regulation is associated with the maintenance of the undifferentiated phenotype of human embryonic stem cells. J Biol Chem. 2010; 285(12):8719-32. PMC: 2838295. DOI: 10.1074/jbc.M109.077081. View

10.

Ng V, Ang S, Chan J, Choo A . Characterization of epithelial cell adhesion molecule as a surface marker on undifferentiated human embryonic stem cells. Stem Cells. 2009; 28(1):29-35. DOI: 10.1002/stem.221. View

11.

Anderson E, Baltus A, Roepers-Gajadien H, Hassold T, de Rooij D, van Pelt A . Stra8 and its inducer, retinoic acid, regulate meiotic initiation in both spermatogenesis and oogenesis in mice. Proc Natl Acad Sci U S A. 2008; 105(39):14976-80. PMC: 2542382. DOI: 10.1073/pnas.0807297105. View

12.

Yoshida S, Takakura A, Ohbo K, Abe K, Wakabayashi J, Yamamoto M . Neurogenin3 delineates the earliest stages of spermatogenesis in the mouse testis. Dev Biol. 2004; 269(2):447-58. DOI: 10.1016/j.ydbio.2004.01.036. View

13.

Brinster R, Zimmermann J . Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A. 1994; 91(24):11298-302. PMC: 45218. DOI: 10.1073/pnas.91.24.11298. View

14.

Shinohara T, Orwig K, Avarbock M, Brinster R . Remodeling of the postnatal mouse testis is accompanied by dramatic changes in stem cell number and niche accessibility. Proc Natl Acad Sci U S A. 2001; 98(11):6186-91. PMC: 33443. DOI: 10.1073/pnas.111158198. View

15.

Yoshinaga K, Ogawa M, Hayashi S, Kunisada T, Fujimoto T, Nishikawa S . Role of c-kit in mouse spermatogenesis: identification of spermatogonia as a specific site of c-kit expression and function. Development. 1991; 113(2):689-99. DOI: 10.1242/dev.113.2.689. View

16.

Kanatsu-Shinohara M, Miki H, Inoue K, Ogonuki N, Toyokuni S, Ogura A . Long-term culture of mouse male germline stem cells under serum-or feeder-free conditions. Biol Reprod. 2004; 72(4):985-91. DOI: 10.1095/biolreprod.104.036400. View

17.

Munz M, Kieu C, Mack B, Schmitt B, Zeidler R, Gires O . The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation. Oncogene. 2004; 23(34):5748-58. DOI: 10.1038/sj.onc.1207610. View

18.

Kanatsu-Shinohara M, Muneto T, Lee J, Takenaka M, Chuma S, Nakatsuji N . Long-term culture of male germline stem cells from hamster testes. Biol Reprod. 2007; 78(4):611-7. DOI: 10.1095/biolreprod.107.065615. View

19.

Kanatsu-Shinohara M, Toyokuni S, Shinohara T . CD9 is a surface marker on mouse and rat male germline stem cells. Biol Reprod. 2003; 70(1):70-5. DOI: 10.1095/biolreprod.103.020867. View

20.

Gonzalez B, Denzel S, Mack B, Conrad M, Gires O . EpCAM is involved in maintenance of the murine embryonic stem cell phenotype. Stem Cells. 2009; 27(8):1782-91. DOI: 10.1002/stem.97. View