Mehmet ÖZBEK, Mustafa ÖZTOP, Emel ERGÜN, Levent ERGÜN, Füsun ERHAN, Banu KANDİL, Feyzullah BEYAZ

Postnatal Gelişim Döneminde Rat Testis ve Epididimisinde Toll-benzeri Reseptörler 2, 7 ve 8’in Ekspresyon Profilleri

Toll-benzeri reseptörler, çeşitli zararlı patojenlerin saldırısına yanıt olarak doğal bağışıklıkta önemli bir rol oynamaktadırlar. Postnatal gelişim boyunca rat testis ve epididimisinde TLR2, 7 ve 8 ifadesini araştırmayı amaçladık. Prepubertal dönemde TLR2 ve 7’nin, testis ve epididimisteki peritübüler miyoid hücreler, interstisyal hücreler, kan damarları, epitel hücreleri ve duktal düz kas hücrelerinde değişik şekillerde lokalize olduğu tespit edildi. Pubertal dönemde TLR2 ve 7 immunboyanması, testiste diğer hücrelerin yanı sıra primer spermatositlerde görülmeye başladı. Epididimiste dar hücreler, güçlü intrasitoplazmik boyanma gösterdi. Postpubertal dönemde farklı gelişimsel aşamalardaki spermatidlerde ortagüçlü derecede TLR2 ve 7 immünboyanması görülürken, pakiten spermatositlerde zayıf immünboyanma görüldü. Testis ve epididimisteki diğer hücre tipleri değişken derecelerde TLR2 ve 7 immünboyanması göstermiştir. Ancak testiste sadece interstisyal hücrelerde zayıf-orta derecede TLR8 immünreaksiyonu saptandı. Erişkin dönemde testis ve epididimisteki farklı hücre tiplerinde TLR2, 7 ve 8 immünboyanmasının artma eğiliminde olduğu dikkat çekti. Bulgularımız TLR2, 7 ve 8 ekspresyonunun postnatal gelişim sürecinde dinamik olarak değiştiğini ve erişkin döneme doğru arttığını göstermektedir. TLR2, 7 ve 8’in postnatal gelişim sürecinde spermatogenezin düzenlenmesi ve testis ile epididimisin doğal bağışıklığının sürdürülmesi ile ilişkili olabileceğini düşünüyoruz.

Expression Profiles of Toll-like Receptors 2, 7 and 8 in Rat Testis and Epididymis During Postnatal Developmental Period

Toll-like receptors take an essential part in innate immunity in response to invasion of the various harmful pathogens. We aimed to investigateTLR2, 7 and 8 expression in rat testis and epididymis throughout postnatal development. In the prepubertal period, TLR2 and 7 were variablylocalized to peritubular myoid cells, interstitial cells, blood vessels, epithelial cells, ductal smooth muscle cells in testis and epididymis. In thepubertal period, immunostaining of TLR2 and 7 started to be seen in primary spermatocytes, as well as other cells, in the testis. Narrow cellsshowed strong intracytoplasmic staining in the epididymis. In the postpubertal period, moderate to strong immunostaining of TLR2 and TLR7 wasseen in spermatids at different developmental steps but weak immunoreaction in pachytene spermatocytes. Other cells in testis and epididymisshowed variable immunostaining of TLR2 and 7. However, weak to moderate immunoreaction to TLR8 was detected in only interstitial cells intestis. In the mature period, immunostaining of TLR2, 7 and 8 tended to increase in different types of cells in testis and epididymis. Our findingssuggest that expression of TLR2, 7 and 8 changed dynamically during postnatal development and increased towards mature period. We considerthat TLR2, 7 and 8 might be associated with the regulation of spermatogenesis and the maintenance of innate immunity of testis and epididymisduring postnatal development.

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1. Bayraktar R, Bertilaccio MTS, Calin GA: The interaction between two worlds: MicroRNAs and toll-like receptors. Front Immunol, 10:1053, 2019. DOI: 10.3389/fimmu.2019.01053
2. McClure R, Massari P: TLR-dependent human mucosal epithelial cell responses to microbial pathogens. Front Immunol, 5:386, 2014. DOI: 10.3389/fimmu.2014.00386
3. Roach JC, Glusman G, Rowen L, Kaur A, Purcell MK, Smith KD, Hood LE, Aderem A: The evolution of vertebrate Toll-like receptors. Proc Natl Acad Sci USA, 102 (27): 9577-9582, 2005. DOI: 10.1073/pnas.0502272102
4. Diebold SS, Kaisho T, Hemmi H, Akira S, Reis e Sousa C: Innate antiviral responses by means of TLR7-mediated recognition of singlestranded RNA. Science, 303 (5663): 1529-1531, 2004. DOI: 10.1126/science. 1093616
5. Heil F, Hemmi H, Hochrein H, Ampenberger F, Kirschning C, Akira S, Lipford G, Wagner H, Bauer S: Species-specific recognition of singlestranded RNA via toll-like receptor 7 and 8. Science, 303 (5663): 1526- 1529, 2004. DOI: 10.1126/science.1093620
6. Lund JM, Alexopoulou L, Sato A, Karow M, Adams NC, Gale NW, Iwasaki A, Flavell RA: Recognition of single-stranded RNA viruses by toll-like receptor 7. Proc Natl Acad Sci U S A, 101 (15): 5598-5603, 2004. DOI: 10.1073/pnas.0400937101
7. O’Neill LAJ, Dinarello CA: The IL-1 receptor/toll-like receptor superfamily: Crucial receptors for inflammation and host defense. Immunol Today, 21 (5): 206-209, 2000. DOI: 10.1016/S0167-5699(00)01611-X
8. Randhawa AK, Hawn TR: Toll-like receptors: Their roles in bacterial recognition and respiratory infections. Expert Rev Anti Infect Ther, 6 (4): 479-495, 2008. DOI: 10.1586/14787210.6.4.479
9. Pandey S, Agrawal DK: Immunology of toll-like receptors: Emerging trends. Immunol Cell Biol, 84 (4): 333-341, 2006. DOI: 10.1111/j.1440- 1711.2006.01444.x
10. Takeda K, Akira S: Toll-like receptors. Curr Protoc Immunol, 109 (1): 14.12.1-14.12.10, 2015. DOI: 10.1002/0471142735.im1412s109
11. Redgrove KA, McLaughlin EA: The role of the immune response in chlamydia trachomatis infection of the male genital tract: A double-edged sword. Front Immunol, 5:534, 2014. DOI: 10.3389/fimmu.2014.00534
12. Haidl G, Allam JP, Schuppe HC: Chronic epididymitis: Impact on semen parameters and therapeutic options. Andrologia, 40 (2): 92-96, 2008. DOI: 10.1111/j.1439-0272.2007.00819.x
13. Schuppe HC, Meinhardt A, Allam JP, Bergmann M, Weidner W, Haidl G. Chronic orchitis: A neglected cause of male infertility? Andrologia, 40 (2): 84-91, 2008. DOI: 10.1111/j.1439-0272.2008.00837.x
14. Biswas B, Narmadha G, Choudhary M, French FS, Hall SH, Yenugu S: Identification of Toll-like receptors in the rat (Rattus norvegicus): Messenger RNA expression in the male reproductive tract under conditions of androgen variation. Am J Reprod Immunol, 62 (4): 243-252, 2009. DOI: 10.1111/j.1600-0897.2009.00732.x
15. Zhao YT, Guo JH, Wu ZL, Xiong Y, Zhou WL: Innate immune responses of epididymal epithelial cells to Staphylococcus aureus infection. Immunol Lett, 119 (1-2): 84-90, 2008. DOI: 10.1016/j.imlet.2008.05.002
16. Palladino MA, Savarese MA, Chapman JL, Dughi MK, Plaska D: Localization of Toll-like receptors on epididymal epithelial cells and spermatozoa. Am J Reprod Immunol, 60 (6): 541–555, 2008. DOI: 10.1111/j.1600-0897.2008.00654.x
17. Liman N, Alan E, Apaydın N: The expression and localization of Toll-like receptors 2, 4, 5 and 9 in the epididymis and vas deferens of an adult tom cats. Theriogenology, 128, 62-73, 2019. DOI: 10.1016/j. theriogenology.2019.02.001
18. Özbek M, Hitit M, Yıldırım N, Özgenç Ö, Ergün E, Ergün L, Beyaz F, Kurtdede N, Altunay H: Expression pattern of galectin-1 and galectin-3 in rat testes and epididymis during postnatal development. Acta Histochem, 120 (8): 814-827, 2018. DOI: 10.1016/j.acthis.2018.09.006
19. Özbek M, Hitit M, Öztop M, Beyaz F, Ergün E, Ergün L: Spatiotemporal expression patterns of natriuretic peptides in rat testis and epididymis during postnatal development. Andrologia, 51 (10): e13387, 2019. DOI: 10.1111/and.13387
20. Boukenaoui-Ferrouk N, Moudilou E, Amirat Z, Exbrayat JM, Khammar F: Morphometric study and immunolocalization of androgen receptors in epididymis during postnatal development in D’Man Lamb reared under arid environment in Algeria. Kafkas Univ Vet Fak Derg, 23 (5): 683-689, 2017. DOI: 10.9775/kvfd.2016.17255
21. Palladino MA, Johnson TA, Gupta R, Chapman JL, Ojha P: Members of the Toll-like receptor family of innate immunity pattern-recognition receptors are abundant in the male rat reproductive tract. Biol Reprod, 76 (6): 958-964, 2007. DOI: 10.1095/biolreprod.106.059410
22. Nishimura M, Naito S: Tissue-specific mRNA expression profiles of human Toll-like receptors and related genes. Biol Pharm Bull, 28 (5): 886- 892, 2005. DOI: 10.1248/bpb.28.886
23. Hack CJ: Integrated transcriptome and proteome data: The challenges ahead. Brief Funct Genomic Proteomic, 3 (3): 212-219, 2004. DOI: 10.1093/ bfgp/3.3.212
24. Cox B, Kislinger T, Emili A: Integrating gene and protein expression data: Pattern analysis and profile mining. Methods, 35 (3): 303-314, 2005. DOI: 10.1016/j.ymeth.2004.08.021
25. Harris CL, Vigar MA, Rey Nores JE, Horejsi V, Labeta MO, Morgan BP: The lipopolysaccharide co-receptor CD14 is present and functional in seminal plasma and expressed on spermatozoa. Immunology, 104 (3): 317-323, 2001. DOI: 10.1046/j.1365-2567.2001.01312.x
26. Mayer C, Adam M, Glashauser L, Dietrich K, Schwarzer JU, Köhn FM, Strauss L, Welter H, Poutanen M, Mayerhofer A: Sterile inflammation as a factor in human male infertility: Involvement of Toll like receptor 2, biglycan and peritubular cells. Sci Rep, 6:37128, 2016. DOI: 10.1038/srep37128
27. Wu H, Wang H, Xiong W, Chen S, Tang H, Han D: Expression patterns and functions of toll-like receptors in mouse sertoli cells. Endocrinology, 149 (9): 4402-4412, 2008. DOI: 10.1210/en.2007-1776