Sher Bahadur KHAN, Zia Ur REHMAN, Qing LIU, Dinesh BHATTARAI, Aixin LIANG, Liguo YANG, Farman ULLAH, Rahim DAD, Hira Sajjad TALPUR, Sher Ali KHAN

Gelişme Dönemindeki Fare Testislerinde Sertoli Hücreleri ve Spermatogonial Hücrelerde Sox5 ve Sox6 Ekspresyonu

SRY gen grubu hayvanlar alemine özgüdür. Erkek embriyoda cinsiyet tayini ve testis gelişimi başta olmak üzere birçok sürece dahildirler. Sox familyası genleri çeşitli hücrelerde belirlenmiş olsa da, Sertoli hücrelerindeki ekspresyon özellikleri ve rolleri tam olarak tanımlanmamıştır. Bu araştırmada, puberta öncesi ve sonrası bir ila sekiz haftalık yaştaki farelerde testis gelişimi sırasında Sertoli hücrelerindeki SoxD grubu Sox5 ve Sox6 genlerinin ekspresyonu araştırıldı. Ekspresyon, tüm testis örneklerinde immünohistokimya ve tüm yaş gruplarının mRNA seviyesini belirlemek için qPCR ile incelenirken, spesifik yaş gruplarında lokalizasyonun belirlenmesi için immünositokimya yapıldı. Sertoli hücrelerinin qPCR sonuçlarında, ilk haftadan sekizinci haftaya kadar farklı ekspresyon seviyeleri gözlendi. Puberta öncesi yaşlarda Sox5’in mRNA seviyesi anlamlı derecede yüksekti (P

Expression Profile of Sox5 and Sox6 in Sertoli and Spermatogonial Cells in Growing Mice Testis

SRY box genes are peculiar to animal kingdom. They are involved in many processes particularly sex determination and testes developmentin male embryo. Although Sox family genes have been identified in various cells, their expression pattern and role is not entirely recognizedin Sertoli cells. In this research, we focused on the expression of SoxD group Sox5 and Sox6 genes in Sertoli cells of mice during pre- and postpubertal testicular development ranging from one-week-old to eight-week-old mice. The expression was studied by immunohistochemistry onwhole testes, and qPCR to determine the mRNA level of all age groups, while immunocyto-chemistry was performed for localization in specificage groups. qPCR results of Sertoli cells from first week to eight week showed diffrent levels of expression. The mRNA level of Sox5, during prepubertal age, was significantly high (P

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1. Chojnacka K, Zarzycka M, Mruk DD: Biology of the sertoli cell in the fetal, pubertal, and adult mammalian testis. Results Probl Cell Diffr, 58, 225- 251, 2016. DOI: 10.1007/978-3-319-31973-5_9
2. Munger SC, Capel B: Sex and the circuitry: Progress toward a systemslevel understanding of vertebrate sex determination. Wiley Interdiscip Rev Syst Biol Med, 4 (4): 401-412, 2012. DOI: 10.1002/wsbm.1172
3. Wilhelm D, Hiramatsu R, Mizusaki H, Widjaja L, Combes AN, Kanai Y, Koopman P: SOX9 regulates prostaglandin D synthase gene transcription in vivo to ensure testis development. J Biol Chem, 282 (14): 10553-10560, 2007. DOI: 10.1074/jbc.M609578200
4. Sargent KM, McFee RM, Spuri Gomes R, Cupp AS: Vascular endothelial growth factor A: Just one of multiple mechanisms for sex-specific vascular development within the testis? J Endocrinol, 227 (2): R31-R50, 2015. DOI: 10.1530/joe-15-0342
5. Lefebvre V, Dumitriu B, Penzo-Mendez A, Han Y, Pallavi B: Control of cell fate and dentiation by Sry-related high-mobility-group box (Sox) transcription factor. Int J Biochem Cell Biol, 39 (12): 2195-2214, 2007. DOI: 10.1016/j.biocel.2007.05.019
6. Wegner M: From head to toes: The multiple facets of Sox proteins. Nucleic Acids Res, 27, 1409-1420, 1999. DOI: 10.1093/nar/27.6.1409
7. Lefebvre V, Li P, de Crombrugghe B: Anew long form of sox5 (L‐Sox5), sox6 and sox 9 are coexpressed in condrogenesis and cooperatively activate thetype II collagen gene. EMBO J, 17, 5718–5733, 1998. DOI: 10.1093/emboj/17.19.5718
8. Sekido R, Lovell-Badge R: Sex determination involves synergistic action of SRY and SF1 on a specific Sox9 enhancer. Nature, 453 (7197): 930-934, 2008. DOI: 10.1038/nature06944
9. Otake T, Kuroiwa A: Molecular mechanism of male diffrentiation is conserved in the SRY-absent mammal, Tokudaia osimensis. Sci Rep, 6:32874, 2016. DOI: 10.1038/srep32874
10. Larney C, Bailey TL, Koopman P: Switching on sex: Transcriptional regulation of the testis-determining gene Sry. Development, 141, 2195- 2205, 2014. DOI: 10.1242/dev.107052
11. Araujo FC, Milsted A, Watanabe IK, Del Puerto HL, Santos RA, Lazar J, Reis FM, Prokop JW: Similarities and diffrences of X and Y chromosome homologous genes, SRY and SOX3, in regulating the renin-angiotensin system promoters. Physiol Genomics, 47 (5): 177-186, 2015. DOI: 10.1152/ physiolgenomics.00138.2014
12. Connor F, Wright E, Denny P, Koopman P, Ashworth A: The sry related HMG box containg gene sox6 is expressed in the adult testis and developing nervous system of the mouse. Nucleic Acids Res, 23, 3365-3372, 1995.
13. Lefebvre V: The SoxD transcription factors--Sox5, Sox6, and Sox13-are key cell fate modulators. Int J Biochem Cell Biol, 42 (3): 429-432, 2010. DOI: 10.1016/j.biocel.2009.07.016
14. Daigle M, Roumaud P, Martin LJ: Expressions of Sox9, Sox5, and Sox13 transcription factors in mice testis during postnatal development. Mol Cell Biochem, 407 (1-2): 209-221, 2015. DOI: 10.1007/s11010-015-2470-7
15. Baroti T, Schillinger A, Wegner M, Stolt CC: Sox13 functionally complements the related Sox5 and Sox6 as important developmental modulators in mouse spinal cord oligodendrocytes. J Neurochem, 136 (2): 316-328, 2016. DOI: 10.1111/jnc.13414
16. Denny P, Swift S, Connor F, Ashworth A: An SRY-related gene expressed during spermatogenesis in the mouse encodes a sequence-specific DNAbinding protein. EMBO J, 10, 3705-3712, 1992.
17. Hersh CP, Silverman EK, Gascon J, Bhattacharya S, Klanderman BJ, Litonjua AA, Lefebvre V, Sparrow D, Reilly JJ, Anderson WH, Lomas DA, Mariani TJ: SOX5 is a candidate gene for chronic obstructive pulmonary disease susceptibility and is necessary for lung development. Am J Respir Crit Care Med, 183 (11): 1482-1489, 2011. DOI: 10.1164/rccm.201010-1751OC
18. Ikeda T, Kawaguchi H, Kamekura S, Ogata N, Mori Y, Nakamura K, Ikegawa S, Chung U: Distinct roles of Sox5, Sox6, and Sox9 in diffrent stages of chondrogenic diffrentiation. J Bone Miner Metab, 23 (5): 337-340, 2005. DOI: 10.1007/s00774-005-0610-y
19. Smits P, Lefebvre V: Sox5 and Sox6 are required for notochord extracellular matrix sheath formation, notochord cell survival and development of the nucleus pulposus of intervertebral discs. Development, 130 (6): 1135-1148, 2003. DOI: 10.1242/dev.00331
20. Murakami A, Ishida S, Thurlow J, Revest JM, Dickson C: SOX6 binds CtBP2 to repress transcription from the Fgf-3 promoter. Nucleic Acids Res, 29 (16): 3347-3355, 2001.
21. Yi Z, Cohen-Barak O, Hagiwara N, Kingsley PD, Fuchs DA, Erickson DT, Epner EM, Palis J, Brilliant MH: Sox6 directly silences epsilon globin expression in definitive erythropoiesis. PLoS Genet, 2 (2): e14, 2006. DOI: 10.1371/journal.pgen.0020014
22. Hagiwara N, Yeh M, Liu A: Sox6 is required for normal fiber type diffrentiation of fetal skeletal muscle in mice. Dev Dyn, 236 (8): 2062-2076, 2007. DOI: 10.1002/dvdy.21223
23. Cohen-Barak O, Hagiwara N, Arlt MF, Horton JP, Brilliant MH: Cloning, characterization and chromosome mapping of the human SOX6 gene. Gene, 265 (1-2): 157-164, 2001. DOI: 10.1016/s0378-1119(01)00346-8
24. Juanchich A, Bardou P, Rue O, Gabillard JC, Gaspin C, Bobe J, Guiguen Y: Characterization of an extensive rainbow trout miRNA transcriptome by next generation sequencing. BMC Genomics, 17:164, 2016. DOI: 10.1186/ s12864-016-2505-9
25. Rudrabhatla P, Utreras E, Jaff H, Kulkarni AB: Regulation of Sox6 by cyclin dependent kinase 5 in brain. PLoS One, 9 (3): e89310, 2014. DOI: 10.1371/journal.pone.0089310
26. Hagiwara N, Klewer SE, Samson RA, Erickson DT, Lyon MF, Brilliant MH: Sox6 is a candidate gene for p100H myopathy, heart block, and sudden neonatal death. Proc Natl Acad Sci U S A, 97 (8): 4180-4185, 2000. DOI: 10.1073/pnas.97.8.4180
27. Ohta S, Misawa A, Lefebvre Vr, Okano H, Kawakami Y, Toda M: Sox6 up-regulation by macrophage migration inhibitory factor promotes survival and maintenance of mouse neural stem/progenitor cells. PLoS One, 8 (9): e74315, 2013. DOI: 10.1371/journal.pone.0074315.g001
28. Rehman ZU, Worku T, Davis JS, Talpur HS, Bhattarai D, Kadariya I, Hua G, Cao J, Dad R, Farmanullah, Hussain T, Yang L: Role and mechanism of AMH in the regulation of sertoli cells in mice. J Steroid Biochem Mol Biol, 174, 133-140, 2017. DOI: 10.1016/j.jsbmb.2017.08.011
29. Chang YF, Lee-Chang JS, Panneerdoss S, MacLean JA, Rao MK: Isolation of sertoli, leydig, and spermatogenic cells from the mouse testis. Biotechniques, 51 (5): 341-342, 344, 2011. DOI: 10.2144/000113764
30. Xiao K, Zou WH, Yang Z, Zia ur R, Ansari AR, Yuan HR, Zhou Y, Cui L, Peng KM, Song H: The role of visfatin on the regulation of inflmmation and apoptosis in the spleen of LPS-treated rats. Cell Tissue Res, 359 (2): 605- 618, 2015. DOI: 10.1007/s00441-014-1997-3
31. Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J: qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol, 8 (2): R19, 2007. DOI: 10.1186/gb-2007-8-2-r19
32. Watanabe M, Kawasaki K, Kawasaki M, Portaveetus T, Oommen S, Blackburn J, Nagai T, Kitamura A, Nishikawa A, Kodama Y, Takagi R, Maeda T, Sharpe PT, Ohazama A: Spatio-temporal expression of Sox genes in murine palatogenesis. Gene Expr Patterns, 21 (2): 111-118, 2016. DOI: 10.1016/j.gep.2016.05.002
33. Stolt CC, Schlierf A, Lommes P, Hillgärtner S, Werner T, Kosian T, Sock E, Kessaris N, Richardson WD, Lefebvre V, Wegner M: SoxD proteins inflence multiple stages of oligodendrocyte development and modulate SoxE protein function. Developmental Cell, 11 (5): 697-709, 2006. DOI: 10.1016/j.devcel.2006.08.011
34. Kiselak EA, Shen X, Song J, Gude DR, Wang J, Brody SL, Strauss JF, Zhang Z: Transcriptional regulation of an axonemal central apparatus gene, sperm-associated antigen 6, by a sry-related high mobility group transcription factor, S-SOX5. J Biol Chem, 285 (40): 30496-30505, 2010. DOI: 10.1074/jbc.M110.121590
35. Ikeda T, Zhang J, Chano T, Mabuchi A, Fukuda A, Kawaguchi H, Nakamura K, Ikegawa S: Identification and characterization of the human long form of Sox5 (L-SOX5) gene. Gene, 298, 59-68, 2002. DOI: 10.1016/ s0378-1119(02)00927-7
36. Takamatsu N, Kanda H, Tsuchiya I, Yamada S, Ito M, Kabeno S, Shiba T, Yamashita S: A gene that is related to SRY and is expressed in the testes encodes a leucine zipper-containing protein. Mol Cell Biol, 15 (7): 3759- 3766, 1995. DOI: 10.1128/mcb.15.7.3759
37. Lefebvre V, Behringer RR, de Crombrugghe B: L-Sox5, Sox6 and Sox9 control essential steps of the chondrocyte diffrentiation pathway. Osteoarthritis Cartilage, 9 (Suppl A): S69-S75, 2001.
38. Grosschedl R, Giese K, Pagel J: HMG domain proteins: architectural elements in the assembly of nucleoprotein structures. Trends Genet, 10 (3): 94-100, 1994. DOI: 10.1016/0168-9525(94)90232-1
39. Hamada-Kanazawa M, Ogawa D, Takano M, Miyake M: Sox6 suppression induces RA-dependent apoptosis mediated by BMP-4 expression during neuronal diffrentiation in P19 cells. Mol Cell Biochem, 412 (1-2): 49-57, 2016. DOI: 10.1007/s11010-015-2607-8
40. Smits P, Li P, Mandel J, Zhang Z, Deng JM, Behringer RR, de Crombrugghe B, Lefebvre V: The transcription factors L-Sox5 and Sox6 are essential for cartilage formation. Dev Cell, 1 (2): 277-290, 2001.