Bıldırcın Testisinde Zonula Okludens-1 ve Klaudin-1 Proteinlerinin Ekspresyonu

Bu çalışmanın amacı, bıldırcınların prepubertal ve postpubertal dönemlerinde testisin genel histolojik yapısını değerlendirmek ve klaudin-1, zonula okludens-1 proteinlerinin varlığını ve yerleşimini belirlemektir. Bu çalışmada 6 adet prepubertal ve postpubertal dönemlerdeki bıldırcınlardan elde edilen testis dokuları kullanıldı. Dokular %10’luk formaldehitte fikse edildi. Rutin doku takibi işleminden geçirildikten sonra parafin bloklar elde edildi. Genel histolojik değerlendirme için Crossman’ın üçlü boyama yöntemi kullanıldı. Klaudin-1 ve Zonula Okludens-1 proteinlerinin gösterimi için sırası ile immünohistokimya ve immünfloresan boyamaları gerçekleştirildi. Prepubertal bıldırcın testislerinde immünreaktivite gözlenmemiştir. Claudin-1’in immünoreaktivitesi postpubertal bıldırcın testislerinde Sertoli hücrelerinde ve spermatogonyumlarda sitoplazmik ve membransel olarak ayırt edilmiştir. Zonula occludens-1’in immünreaktivitesi prepubertal bıldırcın testisinde seminifer tübüllerde gözlenmemiştir. Postpubertal dönemdeki bıldırcın testisinde seminifer tübüllerin bazal bölgesinde immünreaktivite gözlenmiştir. Sonuç olarak diğer kanatlılarda olduğu gibi bıldırcınlarda da prepubertal dönemde kan-testis bariyerinin gelişiminin tamamlanmamış olduğu; postpubertal dönemde ise kan-testis bariyerinin oluştuğu ve bu bariyerin oluşumunda klaudin-1 ve zonula okludens-1 proteinlerinin katıldığı sonucuna ulaşılmıştır.

Anahtar Kelimeler:

Bıldırcın, kan-testis bariyeri, klaudin, zonula okludens

Expression of Zonula Occludens-1 and Claudin-1 Proteins in Japanese Quails Testis

The aim of this study was to evaluate the general histological structure of testis in prepubertal and postpubertal stages of quails and determine the presence and location of claudin-1 and zonula occludens-1 proteins. In this study, testicular tissues obtained from 6 prepubertal and postpubertal stage quails were used. Tissue samples were fixed in 10% formaldehyde and processed for paraffin embedding. Crossman’s triple staining method was used for general histological evaluation. Immunohistochemistry and immunofluorescent staining were performed for the expression of claudin-1 and zonula occludens-1 proteins, respectively. It has also been concluded that the proteins of claudin-1 and zonula occludens-1 do not show immunoreactivity because an active blood–testis barrier is not formed yet in seminiferous tubule epithelium in the testis in the prepubertal period as a result of immunohistochemical andimmunofluorescence stainings but show immunoreactivity in the basal area in seminiferous epithelium with the blood–testis barrier formed in the postpubertal period. Immunoreactivity wasn’t observed in prepubertal quail’s testis. The immunoreactivity of claudin-1 has been distinguished as cytoplasmic and membranous in Sertoli cells and spermatogonia in postpubertal quails’ testis. The immunoreactivity of zonula occludens-1 has not been observed in seminiferous tubules in prepubertal stage quail’s testis. Immunoreactivity has been observed in the basal half of seminiferous tubules in postpubertal stage quail’s testis. It has also been concluded that the proteins of claudin-1 and zonula occludens-1 do not show immunoreactivity because an active blood–testis barrier is not formed yet in seminiferous tubule epithelium in the testis in the prepubertal period as a result of immunohistochemical and immunofluorescence stainings but show immunoreactivity in the basal area in seminiferous epithelium with the blood–testis barrier formed in the postpubertal period.

Keywords:

Blood–testis barrier, claudin, Japanese quail, zonula occludens,

PDF

___

1. Fijak M, Meinhardt A. The testis in immune privilege. Immunol Rev. 2006;213(1):66-81. [CrossRef]
2. Mruk DD, Cheng CY. The mammalian blood-testis barrier: its biology and regulation. Endocr Rev. 2015;36(5):564-591. [CrossRef]
3. Türkmenoğlu İ, Abacıoğlu S. Deney hayvanlarında testis’ in fonksiyonel anatomisi ve embriyolojisi. Turkish Veterinary Journal. 2021;3(1):26-33.
4. Mruk DD, Cheng CY. Sertoli-Sertoli and Sertoli-germ cell interactions and their significance in germ cell movement in the seminiferous epithelium during spermatogenesis. Endocr Rev. 2004;25(5):747-806. [CrossRef]
5. Shouman Z, Marei HE, Abd-Elmaksoud A, et al. Morphological features of the testis among autoimmune mouse model and healthy strains. Microsc Microanal. 2021;27(5):1-9. [CrossRef]
6. Hui L, Nie Y, Li S, et al. Matrix metalloproteinase 9 facilitates Zika virus invasion of the testis by modulating the integrity of the bloodtestis barrier. PLOS Pathog. 2020;16(4):e1008509. [CrossRef]
7. Furuse M, Sasaki H, Fujimoto K, Tsukita S. A single gene product, claudin-1 or-2, reconstitutes tight junction strands and recruits occludin in fibroblasts. J Cell Biol. 1998;143(2):391-401. [CrossRef]
8. Mineta K, Yamamoto Y, Yamazaki Y, et al. Predicted expansion of the claudin multigene family. FEBS Lett. 2011;585(4):606-612. [CrossRef]
9. Günzel D, Yu AS. Claudins and the modulation of tight junction permeability. Physiol Rev. 2013;93(2):525-569. [CrossRef]
10. Otani T, Furuse M. Tight junction structure and function revisited. Trends Cell Biol. 2020;30(10):805-817. [CrossRef]
11. Bhat AA, Syed N, Therachiyil L, et al. Claudin-1, a double-edged sword in cancer. Int J Mol Sci. 2020;21(2):569. [CrossRef]
12. Liman N. The abundance and localization of claudin‐1 and‐5 in the adult tomcats (Felis catus) testis, tubules rectus, rete testis, efferent ductules, and epididymis. Anat Rec. 2023. [CrossRef]
13. Park CJ, Lee JE, Oh YS, et al. Expression of claudin-1 and-11 in immature and mature pheasant (Phasianus colchicus) testes. Theriogenology. 2011;75(3):445-458. [CrossRef]
14. Tsukita S, Furuse M, Itoh M. Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol. 2001;2(4):285-293. [CrossRef]
15. Siti Sarah CO, Nur Husna SM, Md Shukri N, Wong KK, Mohd Ashari NS. Zonula occludens-1 expression is reduced in nasal epithelial cells of allergic rhinitis patients. PeerJ. 2022;10:e13314. [CrossRef]
16. Bauer H, Zweimueller-Mayer J, Steinbacher P, Lametschwandtner A, Bauer HC. The dual role of zonula occludens (ZO) proteins. J Biomed Biotechnol. 2010;2010:402593. [CrossRef]
17. Fanning AS, Van Itallie CM, Anderson JM. Zonula occludens-1 and-2 regulate apical cell structure and the zonula adherens cytoskeleton in polarized epithelia. Mol Biol Cell. 2012;23(4):577-590. [CrossRef]
18. Ram AK, Vairappan B. Role of zonula occludens in gastrointestinal and liver cancers. World J Clin Cases. 2022;10(12):3647-3661. [CrossRef]
19. Crossmon G. A modification of Mallory's connective tissue stain with a discussion of the principles involved. Anat Rec. 1937;69(1):33-38. [CrossRef]
20. Ozaydin T, Sur E, Oznurlu Y, Celik I, Uluisik D. Immunohistochemical distribution of heat shock protein 70 and proliferating cell nuclear antigen in mouse placenta at different gestational stages. Microsc Res Tech. 2016;79(4):251-257. [CrossRef]
21. Bölükbaş F, Öznurlu Y. Determining the effects of in ovo administration of monosodium glutamate on the embryonic development of brain in chickens. NeuroToxicology. 2023;94:87-97.[CrossRef]
22. DasdelenD, Solmaz M, Menevse E, Mogulkoc R, BaltaciAK, Erdogan E. Increased apoptosis, tumor necrosis factor-α, and DNA damage attenuated by 3′, 4′-dihydroxyflavonol in rats with brain ischemiareperfusion. Indian J Pharmacol. 2021;53(1):39-49. [CrossRef]
23. Hodges RD. The Histology of the Fowl. Cambridge: Academic Press; 1974:300-325.
24. Mruk DD, Cheng CY. Tight junctions in the testis: new perspectives. Philos Trans R Soc Lond B Biol Sci. 2010;365(1546):1621-1635. [CrossRef]
25. Venditti M, Ben Rhouma MB, Romano MZ, Messaoudi I, Reiter RJ, Minucci S. Evidence of melatonin ameliorative effects on the bloodtestis barrier and sperm quality alterations induced by cadmium in the rat testis. Ecotoxicol Environ Saf. 2021;226:112878. [CrossRef]
26. Huang W, Liu M, Xiao B, et al. Aflatoxin b1 disrupts blood-testis barrier integrity by reducing junction protein and promoting apoptosis in mice testes. Food Chem Toxicol. 2021;148:111972. [CrossRef]
27. Molele RA, Mahdy MAA, Zakariah M, Ibrahim MIA, Fosgate GT, Brown G. Age-related histomorphometric and ultrastructural changes in the Sertoli cells of Japanese quail (Coturnix coturnix japonica) Tissue Cell. 2021;73:101650. [CrossRef]
28. Bergmann M, Schindelmeiser J, , Lameu , , . Development of the blood-testis barrier in the domestic fowl (Gallus domesticus). Int J Androl. 1987;10(2):481-488. [CrossRef]
29. Osman DI, Ekwall H, Plöen L. Specialized cell contacts and the blood‐testis barrier in the seminiferous tubules of the domestic fowl (Gallus domesticus) Int J Androl. 1980;3(1-6):553-562.[CrossRef]
30. Gilio JM, Portaro FC, Borella MI, Lameu C, Camargo AC, Alberto-Silva C. A bradykinin potentiating peptide (BPP-10c) from bothrops jararaca induces changes in seminiferous tubules. J Venom Anim Toxins Incl Trop Dis. 2013;19(1):28.[CrossRef]
31. Karateke H. Ratlarda postnatal dönemde testis dokusu ile kan testis bariyerinin gelişiminin histomorfometrik ve immunohistokimyasal değerlendirilmesi.Tez. Afyon Kocatepe Üniversitesi, Sağlık Bilimleri Enstitüsü. 2013.
32. Molele RA, Ibrahim MIA, Zakariah M, et al. Junctional complexes of the blood-testis barrier in the Japanese quail (Coturnix coturnix japonica) Acta Histochem. 2022;124(7):151929. [CrossRef]
33. Gilula NB, Fawcett DW, Aoki A. The Sertoli cell occluding junctions and gap junctions in mature and developing mammalian testis. Dev Biol. 1976;50(1):142-168. [CrossRef]
34. Stevenson BR, Siliciano JD, Mooseker MS, Goodenough DA. Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia. J Cell Biol. 1986;103(3):755-766. [CrossRef]
35. Fink C, Weigel R, Hembes T, et al. Altered expression of ZO-1 and ZO-2 in Sertoli cells and loss of blood-testis barrier integrity in testicular carcinoma in situ. Neoplasia. 2006;8(12):1019-1027. [CrossRef]
36. Byers S, Graham R, Dai HN, Hoxter B. Development of Sertoli cell junctional specializations and the distribution of the tight‐junction‐associated protein ZO‐1 in the mouse testis. Am J Anat. 1991;191(1):35-47. [CrossRef]