N-WASP Is Required for Structural Integrity of the Blood-Testis Barrier

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Mammalian spermatogenesis takes place within a sheltered environment, whereby somatic Sertoli cells protect and guide germ cells as they mature and differentiate. A key structure generated by the protective Sertoli cell epithelium is the blood-testis barrier (BTB), a composite of junctional and cytoskeletal elements, which prevents exposure of post-meiotic spermatids to the immune system. The BTB is a highly dynamic structure, which needs to be dismantled and rapidly rebuilt, in order to allow passage of maturing preleptotene spermatocytes, without compromising their isolation. Here we show that N-WASP, a conserved facilitator of formation of branched actin microfilament arrays, provides a function that is essential for maintenance of an intact BTB. Genetic disruption of N-WASP in mouse Sertoli cells leads to loss of BTB impermeability, resulting in a complete arrest of spermatogenesis at early and post-meiotic stages. Based on the localization patterns of key elements, we propose that branched-actin filaments participate in recycling of BTB materials to ensure the dynamic and efficient maintenance of this structure, one of a series of blood-tissue barriers that preserve privileged organ environments.

Vyšlo v časopise: N-WASP Is Required for Structural Integrity of the Blood-Testis Barrier. PLoS Genet 10(6): e32767. doi:10.1371/journal.pgen.1004447
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1004447

Souhrn

Zdroje

1. de Kretser DM, Kerr JB (1988) The cytology of the testis. In: Knobil E, Neill JB, Ewing LL, Greenwald GS, Markert CL, et al.., editors. The Physiology of Reproduction Vol 1. New York: Raven Press. pp. 837–932.

2. ChengCY, MrukDD (2012) The blood-testis barrier and its implication in male contraception. Pharmacol Rev 64: 16–64.

3. FrancaLR, AuharekSA, HessRA, DufourJM, HintonBT (2012) Blood-tissue barriers: Morphofunctional and immunological aspects of the blood-testis and blood-epididymal barriers. Adv Exp Med Biol 763: 237–259.

4. VoglAW, VaidKS, GuttmanJA (2008) The Sertoli cell cytoskeleton. Adv Exp Med Biol 636: 186–211.

5. RussellLD, PetersonRN (1985) Sertoli cell junctions: morphological and functional correlates. Int Rev Cytol 94: 177–211.

6. ChengCY, MrukDD (2010) A local autocrine axis in the testes that regulates spermatogenesis. Nature Rev Endocrinol 6: 380–395.

7. WangCQF, ChengCY (2007) A seamless trespass: germ cell migration across the seminiferous epithelium during spermatogenesis. J Cell Biol 178: 549–556.

8. RussellLD (1977) Movement of spermatocytes from the basal to the adluminal compartment of the rat testis. Am J Anat 148: 313–328.

9. ChengCY, LiePPY, MokKW, ChengYH, WongEWP, et al. (2011) Interactions of laminin β3 fragment with β1-integrin receptor: a revisit of the apical ES-BTB-hemidesmosome functional axis in the testis. Spermatogenesis 1: 174–185.

10. LiePPY, ChanAYN, MrukDD, LeeWM, ChengCY (2010) Restricted Arp3 expression in the testis prevents blood-testis barrier disruption during junction restructuring at spermatogenesis. Proc Natl Acad Sci USA 107: 11411–11416.

11. KurisuS, TakenawaT (2009) The WASP and WAVE family proteins. Genome Biol 10: 226.221–226.229.

12. PadrickSB, RosenMK (2010) Physical mechanisms of signal integration by WASP family proteins. Annu Rev Biochem 79: 707–735.

13. BurianekLE, SoderlingSH (2013) Under lock and key: Spatiotemporal regulation of WASP family proteins coordinates separate dynamic cellular processes. Semin Cell Dev Biol 24: 258–266.

14. ChengCY, MrukDD (2011) Regulation of spermiogenesis, spermiation and blood-testis barrier dynamics: novel insights from studies on Eps8 and Arp3. Biochem J 435: 553–562.

15. RotkopfS, HambergY, AigakiT, SnapperSB, ShiloBZ, et al. (2011) The WASp-based actin polymerization machinery is required in somatic support cells for spermatid maturation and release. Development 138: 2729–2739.

16. Cotta-de-AlmeidaV, WesterbergL, MaillardMH, OnaldiD, WachtelH, et al. (2007) Wiskott Aldrich syndrome protein (WASP) and N-WASP are critical for T cell development. Proc Natl Acad Sci USA 104: 15424–15429.

17. BitgoodMJ, ShenL, McMahonAP (1996) Sertoli cell signaling by Desert hedgehog regulates the male germline. Curr Biol 6: 298–304.

18. JaegleM, GhazviniM, MandemakersW, PiirsooM, DriegenS, et al. (2003) The POU proteins Brn-2 and Oct-6 share important functions in Schwann cell development. Genes Dev 17: 1380–1391.

19. LindeboomF, GillemansN, KarisA, JaegleM, MeijerD, et al. (2003) A tissue-specific knockout reveals that Gata1 is not essential for Sertoli cell function in the mouse. Nucleic Acids Res 31: 5405–5412.

20. KochM, OlsonP, AlbusA, JinW, HunterD, et al. (1999) Characterization and expression of the laminin γ3 chain: a novel, non-basement membrane-associated, laminin chain. J Cell Biol 145: 605–618.

21. YanHHN, ChengCY (2006) Laminin α3 forms a complex with β3 and γ3 chains that serves as the ligand for α6β1-integrin at the apical ectoplasmic specialization in adult rat testes. J Biol Chem 281: 17286–17303.

22. OatleyJM, BrinsterRL (2006) Spermatogonial stem cells. Methods Enzymol 419: 259–282.

23. van BragtMP, Roepers-GajadienHL, KorverCM, BogerdJ, OkudaA, et al. (2008) Expression of the pluripotency marker UTF1 is restricted to a subpopulation of early A spermatogonia in rat testis. Reproduction 136: 33–40.

24. SharpeRM, McKinnellC, KivlinC, FisherJS (2003) Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. Reproduction 125: 769–784.

25. LiMWM, XiaW, MrukDD, WangCQF, YanHHY, et al. (2006) TNFα reversibly disrupts the blood-testis barrier and impairs Sertoli-germ cell adhesion in the seminiferous epithelium of adult rat testes. J Endocrinol 190: 313–329.

26. MokKW, MrukDD, LeeWM, ChengCY (2012) Spermatogonial stem cells alone are not sufficient to re-initiate spermatogenesis in the rat testis following adjudin-induced infertility. Int J Androl 35: 86–101.

27. CyrDG (2011) Connexins and pannexins: Coordinating cellular communication in the testis and epididymis. Spermatogenesis 1: 325–338.

28. LiMWM, MrukDD, ChengCY (2012) Gap junctions and blood-tissue barriers. Adv Exp Med Biol 763: 260–280.

29. MorrowCMK, MrukDD, ChengCY, HessRA (2010) Claudin and occludin expression and function in the seminiferous epithelium. Philos Trans R Soc Lond B Biol Sci 365: 1679–1696.

30. SiuER, WongEWP, MrukDD, PortoCS, ChengCY (2009) Focal adhesion kinase is a blood-testis barrier regulator. Proc Natl Acad Sci USA 106: 9298–9303.

31. CumminsPM (2012) Occludin: one protein, many forms. Mol Cell Biol 32: 242–250.

32. LeeNPY, MrukDD, LeeWM, ChengCY (2003) Is the cadherin/catenin complex a functional unit of cell-cell-actin-based adherens junctions (AJ) in the rat testis? Biol Reprod 68: 489–508.

33. WangCQF, MrukDD, LeeWM, ChengCY (2007) Coxsackie and adenovirus receptor (CAR) is a product of Sertoli and germ cells in rat testes which is localized at the Sertoli-Sertoli and Sertoli-germ cell interface. Exp Cell Res 313: 1373–1392.

34. Ozaki-KurodaK, NakanishiH, OhtaH, TanakaH, KuriharaH, et al. (2002) Nectin couples cell-cell adhesion and the actin scaffold at heterotypic testicular junctions. Curr Biol 12: 1145–1150.

35. PalombiF, SalanovaM, TaroneG, FariniD, StefaniniM (1992) Distribution of b1 integrin subunit in rat seminiferous epithelium. Biol Reprod 47: 1173–1182.

36. SiuMKY, ChengCY (2004) Interactions of proteases, protease inhibitors, and the β1 integrin/laminin γ3 protein complex in the regulation of ectoplasmic specialization dynamics in the rat testis. Biol Reprod 70: 945–964.

37. XiaoX, ChengCY, MrukDD (2013) Intercellular adhesion molecule-2 is involved in apical ectoplasmic specialization dynamics during spermatogenesis in the rat. J Endocr 216: 73–86.

38. PoukkulaM, KremnevaE, SeerlachiusM, LappalainenP (2011) Actin-depolymerizating factor homology domain: a conserved fold performing diverse roles in cytoskeletal dynamics. Cytoskeleton 68: 471–490.

39. LiMWM, XiaoX, MrukDD, LamYL, LeeWM, et al. (2011) Actin binding protein drebrin E is involved in junction dynamics during spermatogenesis. Spermatogenesis 1: 123–136.

40. AroldST (2011) How focal adhesion kinase achieves regulation by linking ligand binding, localization and action. Curr Opin Struct Biol 21: 808–813.

41. LiePPY, MrukDD, MokKW, SuL, LeeWM, et al. (2012) Focal adhesion kinase-Tyr407 and -Tyr397 exhibit antagonistic effects on blood-testis barrier dynamics in the rat. Proc Natl Acad Sci USA 109: 12562–12567.

42. SiuMKY, MrukDD, LeeWM, ChengCY (2003) Adhering junction dynamics in the testis are regulated by an interplay of β1-integrin and focal adhesion complex (FAC)-associated proteins. Endocrinology 144: 2141–2163.

43. ChengCY, LiePPY, WongEWP, MrukDD (2013) Focal adhesion kinase and actin regulatory/binding proteins that modulate F-actin organization at the tissue barrier. Lession from the testis. Tissue Barriers 1: e24252 (DOI:10.4161/tisb.24252)..

44. MonastyrskaI, HeC, GengJ, HoppeAD, LiZ, et al. (2008) Arp2 links autophagic machinery with the actin cytoskeleton. Mol Biol Cell 19: 1962–1975.

45. ShivasJM, SkopAR (2012) Arp2/3 mediates early endosome dynamics necessary for the maintenance of PAR asymmetry in Caenorhabditis elegans. Mol Biol Cell 23: 1917–1927.

46. YamamotoT, MochidaJ, KadotaJ, TakedaMB, E., TanakaK (2010) Initial polarized bud growth by endocytic recycling in the absence of actin cable-dependent vesicle transport in yeast. Mol Biol Cell 21: 1237–1252.

47. JaegleM, GhazviniM, MandemakersW, PiirsooM, DriegenS, et al. (2003) The POU proteins Brn-2 and Oct-6 share important functions in Schwann cell development. Genes Dev 17: 1380–1391.

48. SnapperSB, TakeshimaF, AntonI, LiuCH, ThomasSM, et al. (2001) N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility. Nature Cell Biol 3: 897–904.

49. XiaoX, MrukDD, LeeWM, ChengCY (2011) c-Yes regulates cell adhesion at the blood-testis barrier and the apical ectoplasmic specialization in the seminiferous epithelium of rat testes. Int J Biochem Cell Biol 43: 651–665.

50. MrukDD, ChengCY (2011) Enhanced chemiluminescence (ECL) for routine immunoblotting. An inexpensive alternative to commercially available kits. Spermatogenesis 1: 121–122.

51. XiaoX, MrukDD, ChengCY (2013) c-Yes regulates cell adhesion at the apical ectoplasmic specialization-blood-testis barrier axis via its effects on protein recruitment and distribution. Am J Physiol Endocrinol Metab 304: E145–E159.

52. LiePPY, MrukDD, LeeWM, ChengCY (2009) Epidermal growth factor receptor pathway substrate 8 (Eps8) is a novel regulator of cell adhesion and the blood-testis barrier integrity in the seminiferous epithelium. FASEB J 23: 2555–2567.