Comparative Expression Analysis of Spermatogonial Stem Cell Markers in Cattle and Sheep
الموضوعات :F. Nasri Ahangar 1 , M. Zandi 2 , M.R. Sanjabi 3 , A. Ghaedrahmati 4
1 - Department of Agriculture, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran
2 - Department of Agriculture, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran
3 - Department of Agriculture, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran
4 - Department of Animal Science, Agricultural Science and Natural Resources University of Khuzestan, Mollasani, Khuzestan, Iran
الکلمات المفتاحية: Molecular markers, pluripotency, cell specific Factor, surface markers, transcrip-tion factor,
ملخص المقالة :
Spermatogenesis is supported by stem cells called spermatogonial stem cells (SSCs), which are capable of transmitting information to the next generation. However, little is known on the specific markers of SSCs in farm animals. We investigated the expression of cdh1, cmyc, bcl6b, plzf, gfra1, nanog, vasa, thy1 and uchl1, as specific markers of SSCs, in bovine and ovine SSCs. The expression of the studied genes was conducted by real-time polymerase chain reaction method. For this reason, the enzymatic digestion process for two times was used to achieve SSCs from male calves and ram lambs testes. Then, filtration and differential plating techniques increased the SSC number in the cell suspension caused by mechanical and enzymatic digestions. Sertoli cells treated with Mitomycin-C were applied to obtain the feeder layer. Culturing of the stem cells was done on Sertoli cell feeder layer. Our results revealed that the expression of nanog and plzf was similar in bovine and ovine SSCs. Unlike bovine SSC colonies, cdh1 gene was not expressed in colony of ovine SSCs and its use as a specific marker of sheep SSCs is not suggested. The expression of uchl, vasa, thy1 and cdh1 genes was significantly higher in bovine SSCs and the bcl6b and cmyc expression was significantly higher in ovine SSCs compared to each other (P<0.05). From the results of this study nanog, plzf, uchl1, vasa, thy1 and cdh1 are suggested as markers of bovine and nanog, plzf, bcl6b and cmyc genes as markers of ovine SSCs.
Abbasi H., Tahmoorespur M., Hosseini S.M., Nasiri Z., Bahadorani M., Hajian M. and Nasr-Esfahani M.H. (2013). THY1 as a reliable marker for enrichment of undifferentiated spermatogonia in the goat. Theriogenology. 80, 923-932.
Aoi T., Yae K., Nakagawa M., Ichisaka T., Okita K., Takahashi K. and Yamanaka S. (2008). Generation of pluripotent stem cells from adult mouse liver and stomach cells. Science. 321, 699-702.
Bahadorani M., Hosseini S.M., Abedi P., Hajian M., Afrough M., Azhdari Tafti Z. and Nasr-Esfahani M.H. (2011). Comparative immunohistochemical analysis of VASA, PLZF and THY1 in goats and sheep suggests that these markers are also conserved in these species. J. Cytol. Histol. 2, 126-133.
Bojnordi M.N., Azizi H., Skutella T., Movahedin M., Pourabdolhossein F., Shojaei A. and Hamidabadi H.G. (2017). Differentiation of spermatogonia stem cells into functional mature neurons characterized with differential gene expression. Mol. Neurobiol. 54, 5676-5682.
Borjigin U., Davey R., Hutton K. and Herrid M. (2010). Expression of promyelocytic leukaemia zinc-finger in ovine testis and its application in evaluating the enrichment efficiency of differential plating. Reprod. Fertil. Dev. 22, 733-742.
Buaas F.W., Kirsh A.L., Sharma M., McLean D.J., Morris J.L., Griswold M.D. and Braun R.E. (2004). Plzf is required in adult male germ cells for stem cell self-renewal. Nat. Genet. 36, 647-652.
Buageaw A., Sukhwani M., Ben-Yehudah A., Ehmcke J., Rawe V.Y., Pholpramool C. and Schlatt S. (2005). GDNF family receptor alpha1 phenotype of spermatogonial stem cells in immature mouse testes. Biol. Reprod. 73, 1011-1016.
Cai Y., Deng M., Liu Z., Zhang G., Pang J., An S. and Wang F. (2020). EZH2 expression and its role in spermatogonial stem cell self-renewal in goats. Theriogenology. 155, 222-231.
Castrillon D.H., Quade B.J., Wang T.Y., Quigley C. and Crum C.P. (2000). The human VASA gene is specifically expressed in the germ cell lineage. Proc. Natl. Acad. Sci. USA. 97, 9585-9590.
Corbineau S., Lassalle B., Givelet M., Souissi-Sarahoui I., Firlej V., Romeo P.H. and Fouchet P. (2017). Spermatogonial stem cells and progenitors are refractory to reprogramming to pluripotency by the transcription factors Oct3/4, c-Myc, Sox2 and Klf4. Oncotarget. 8, 10050-10059.
Fujihara M., Kim S.M., Minami N., Yamada M. and Imai H. (2011). Characterization and in vitro culture of male germ cells from developing bovine testis. J. Reprod. Dev. 157(3), 355-364.
Goel S., Fujihara M., Minami N., Yamada M. and Imai H. (2008). Expression of NANOG, but not POU5F1, points to the stem cell potential of primitive germ cells in neonatal pig testis. Reproduction. 135, 785-795.
Heidari B., Rahmati-Ahmadabadi M., Akhondi M.M., Zarnani A.H., Jeddi-Tehrani M., Shirazi A. and Behzadi B. (2012). Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9. 5 markers. J. Assist. Reprod. Genet. 29, 1029-1038.
Herrid M., Davey R.J. and Hill J.R. (2007). Characterization of germ cells from pre-pubertal bull calves in preparation for germ cell transplantation. Cell Tissue Res. 330, 321-329.
Herrid M., Olejnik J., Jackson M., Suchowerska N., Stockwell S., Davey R. and Hill J.R. (2009). Irradiation enhances the efficiency of testicular germ cell transplantation in sheep. Biol. Reprod. 81, 898-905.
Hoei‐Hansen C.E., Almstrup K., Nielsen J.E., BraskSonne S., Graem N., Skakkebaek N.E. and Rajpert‐De Meyts E. (2005). Stem cell pluripotency factor NANOG is expressed in human fetal gonocytes, testicular carcinoma in situ and germ cell tumours. Histopathology. 47, 48-56.
Izadyar F., Matthijs‐Rijsenbilt J.J., Den Ouden K., Creemers L.B., Woelders H. and de Rooij D.G. (2002). Development of a cryopreservation protocol for type A spermatogonia. J. Androl. 23, 537-545.
Jafarnejad A., Zandi M., Aminafshar M., Sanjabi M.R. and Emamjomeh Kashan N. (2018). Evaluating bovine sperm transfection using a high-performance polymer reagent and assessing the fertilizing capacity of transfected spermatozoa using an in vitro fertilization technique. Arch. Anim. Breed. 61, 351-358.
Kanatsu-Shinohara M., Tanaka T., Ogonuki N., Ogura A., Morimoto H., Cheng P.F. and Shinohara T. (2016). Myc/Mycn-mediated glycolysis enhances mouse spermatogonial stem cell self-renewal. Genes Dev. 30, 2637-2648.
Lee G.S., Kim H.S., Lee S.H., Kang M.S., Kim D.Y., Lee C.K. and Hwang W.S. (2005). Characterization of pig vasa homolog gene and specific expression in germ cell lineage. Mol. Reprod. Dev. 72, 320-328.
Lee K.H., Lee W.Y., Kim J.H., Yoon M.J., Kim N.H., Kim J.H. and Song H. (2013). Characterization of GFR α‐1‐Positive and GFR α‐1‐Negative Spermatogonia in Neonatal Pig Testis. Reprod. Domest. Anim. 48, 954-960.
Livak K.J. and Schmittgen T.D. (2001). Analysis of relative gene expression data using real time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25, 402-408.
McMillan M., Andronicos N., Davey R., Stockwell S., Hinch G. and Schmoelzl S. (2014). Claudin-8 expression in sertoli cells and putative spermatogonial stem cells in the bovine testis. Reprod. Fertil. Dev. 26, 633-644.
Mitchell R.T., Cowan G., Morris K.D., Anderson R.A., Fraser H.M., Mckenzie K.J. and Sharpe R.M. (2008). Germ cell differentiation in the marmoset (Callithrix jacchus) during fetal and neonatal life closely parallels that in the human. Hum. Reprod. 23, 2755-2765.
Nazm Bojnordi M., Movahedin M., Tiraihi T. and Javan M. (2013). Alteration in genes expression patterns during in vitro differentiation of mouse spermatogonial cells into neuroepithelial-like cells. Cytotechnology. 65, 97-104.
Niu B., Li B., Wu C., Wu J., Yan Y., Shang R. and Hua J. (2016). Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells. Oncotarget. 7, 77532-77542.
Oatley J.M., Avarbock M.R., Telaranta A.I., Fearon D.T. and Brinster R.L. (2006). Identifying genes important for spermatogonial stem cell self-renewal and survival. Proc. Natl. Acad. Sci. USA. 103, 9524-9529.
Reding S.C., Stepnoski A.L., Cloninger E.W. and Oatley J.M. (2010). THY1 is a conserved marker of undifferentiated spermatogonia in the pre-pubertal bull testis. Reproduction. 139, 893-903.
Rege T.A. and Hagood J.S. (2006). Thy‐1 as a regulator of cell‐cell and cell‐matrix interactions in axon regeneration, apoptosis, adhesion, migration, cancer, and fibrosis. FASEB. J. 20, 1045-1054.
Rodriguez-Sosa J.R., Dobson H. and Hahnel A. (2006). Isolation and transplantation of spermatogonia in sheep. Theriogenology. 66, 2091-2103.
SPSS Inc. (2011). Statistical Package for Social Sciences Study. SPSS for Windows, Version 20. Chicago SPSS Inc., USA.
Tokuda M., Kadokawa Y., Kurahashi H. and Marunouchi T. (2007). CDH1 is a specific marker for undifferentiated spermatogonia in mouse testes. Biol. Reprod. 76, 130-141.
Toyooka Y., Tsunekawa N., Takahashi Y., Matsui Y., Satoh M. and Noce T. (2000). Expression and intracellular localization of mouse Vasa-homologue protein during germ cell development. Mech. Dev. 93, 139-149.
Wu J., Song W., Zhu H., Niu Z., Mu H., Lei A. and Hua J. (2013). Enrichment and characterization of Thy1-positive male germline stem cells (mGSCs) from dairy goat (Capra hircus) testis using magnetic microbeads. Theriogenology. 80, 1052-1060.
Wu Y., Guo T., Li J., Niu C., Sun W., Zhu S. and Yue Y. (2022). The transcriptional cell atlas of testis development in sheep at pre-sexual maturity. Mol. Biol. 44, 483-497.
Yu X., Riaz H., Dong P., Chong Z., Luo X., Liang A. and Yang L. (2014). Identification and IVC of spermatogonial stem cells in prepubertal buffaloes. Theriogenology. 81, 1312-1322.
Zeeman A.M., Stoop H., Boter M., Gillis A.J., Castrillon D.H., Oosterhuis J.W. and Looijenga L.H. (2002). VASA is a specific marker for both normal and malignant human germ cells. Lab. Invest. 82, 159-166.
Zhang Y., Sachula W.U., Luo F.H., Liu L.H., HU T.Y., Boyang Y.U. and WU Y.J. (2014). CDH1, a novel surface marker of spermatogonial stem cells in sheep testis. J. Integr. Agric. 13, 1759-1765.
Zheng Y., Zhang Y., Qu R., He Y., Tian X. and Zeng W. (2014a). Spermatogonial stem cells from domestic animals: Progress and prospects. Reproduction. 147, 65-74.
Zheng Y., He Y., An J., Qin J., Wang Y., Zhang Y. and Zeng W. (2014b). THY1 is a surface marker of porcine gonocytes. Reprod. Fertil. Dev. 26, 533-539.
