Apoptosis of Germ Cells after Vasectomy in Rats

수컷 랫드에서 졍관절제술에 의한 생식세포의 Apoptosis

  • Choi, Jong-yun (College of Veterinary Medicine, Chungnam National University) ;
  • Cho, Sung-whan (College of Veterinary Medicine, Chungnam National University) ;
  • Ryu, Si-yoon (College of Veterinary Medicine, Chungnam National University) ;
  • Jee, Young-heun (Department of Veterinary Medicine, Cheju National University) ;
  • Lee, Geun-jwa (Chungnam Livestock & Veterinary Service Institute) ;
  • Son, Hwa-young (College of Veterinary Medicine, Chungnam National University)
  • Accepted : 2003.08.02
  • Published : 2003.09.25

Abstract

The pathological mechanism of impaired spermatogenesis after vasectomy has not been completely investigated. In this study, we examined pathological changes of the testis and the Fas-Fas ligand (FasL) mediated signaling pathway in apoptotic germ cell death after vasectomy in rats. Ten-weeks old Sprague-Dawley rats were underwent bilateral vasectomy and sacrificed after 1 day, 2 days, 3 days, 5 days, 1 week, 2 weeks, and 4 weeks of surgery and the testes were removed. Histopathological evaluation of spermatogenesis was performed by hematoxylin-eosin and periodic acid-Schiff-hematoxylin staining. To elucidate the pathophysiology of seminiferous tubule damage, terminal dUTP nick end labeling staining, electrophoresis assay of DNA fragmentation, and Western blotting analysis for Fas-FasL were performed. Relative weights of testes were decreased from 5 days after vasectomy. Germ cell degeneration were first found in the spermatogonia and spermatocytes at stages I-VI, and XII-XIV seminiferous tubules. Mean incidence of apoptotic germ cells after vasectomy progressively increased to peak in 5 days, and then gradually decreased to the control levels in 2 weeks after vasectomy. The expression of Fas-FasL reached maximum level at 5 days after vasectomy and then declined. In conclusion, impaired spermatogenesis after vasectomy associated with an increase in germ cell apoptasis, which is partly mediated by the activation of Fas-FasL.

Keywords

References

  1. Allan, D., Hannon, B. and Kerr, J. Cell death inspermatogenesis. In Perspecdves on Mainmalian CellDeath. PoKen, C. (ed), PP. 229-258. Oxford UniversityPress, London, 1987
  2. Blanco-Rodriguez, J. and Martinez-Garcia, C.Spontaneous genn cell death in the testis of the adultrat takes the form of apoptosis: re-evaluation of celltypes that exhitat ths ability to de durmg spamatogaiesis.Cell Prolif. 1996, 29, 13-31
  3. Boekelheide, K., Fleming, S. L., Johnson, K. J.,Patel, S. R. and Schoenfeld, H. A. Role of Sertolicells in injury-associated testicular germ cell apoptosis.Proc. Soc. Exp. Biol. Med 2000, 225, 105-115
  4. Cai, L., Hales, B. F. and Robaire, B. Induction ofapoptosis in the germ cells of adult male rats exposureto cycloplxBFhanide. Hol. Repod 1997, 56, 1490-1497
  5. De Rooij, D. G. Proliferation and differentiation ofspennatogonial stem cells. Reproduction. 2001, 121,347-354
  6. Dunkel, L., Taskinen, S., Hovatta, 0., TUly, J. andWikstrom, S. Genn cell apoptosis after treatment ofcryptorchidism with human chorionic gonadotropm isassociated with inipaired reproductive funcdon in theadult. J. Clin. Invest., 1997, 100, 2341-2346
  7. Eddy, E. M. Male germ cell gene expression. RecentProg. Horm. Res. 2002, 57, 103-128
  8. Ejima, K., Kqji, T., Tsuruta, D., Nanri, H.,Kashiinura, M. and Ikeda, M. Induction of apoptosisin placentas of pregnant mice exposed tolipopoly-saccharides: possible involvement of Pas/Fas ligandsystem. Biol. Reprod. 2000, 62, 178-185
  9. Friesen, C., Herr, L, Krammer, P. H. and Debatin,K. M. hivolvement of the CD95 (APO-1/Fas) receptor/ligand system in drug-induced apoptosis in leukemiacells. Nat. Med. 1996, 2, 574-577
  10. Grossfeld, G. D. and Small, E. J. Long-term sideeffects of treatment for testis cancer. Urol. Qin. North.Am. 1998, 25, 503-515
  11. Hakuno, N., Koji, T., Yano, T., Kobayashi, N.,Tsutsumi, O., Taketani, Y. and Nakane, P. K.Fas/AP0-1/CD95 system as a mediator of granulosacell apoptosis in ovaiian follicle atresia. Endocrin. 1996,137, 1398-1348
  12. Henriksens, K., Hakovirta, H. and Parvinen, M. Insitu quaniification of stage specific apoptosis in the rat seniniferous epithelium effecls of short-term experimental cryptorchidism. Int. J. Androl. 1995, 18, 256-262
  13. Higami. Y., Shiinokawa, I., Tomita, M., Okimoto, T., Koji, T., Kobayashi, N. and Ikeda, T. Aging accelerates but life-long dietary restriction suppresses apoptosis related Fas expression on hepatocytes. Am. J. Pathol. 1997, 151, 659-663
  14. Hikim, A. P., Wang, C., Leung, A. and Swerdloff, R. S. Involvement of apoptosis in the induction of germ cell degeneration in adult rats after gonadotropin-releasing hormone antagonist treatment. Endocrinology.1995, 136, 2770-2775
  15. Iwamoto, M., Koji, T., Nlakiyania, K., Kubayashi,N. and Nakane, P. K. Apoptosis of crypt epithelialcells in ulcerative coUtis. J. Pathol. 1996, 180, 152-259
  16. Kawakaini, A., Eguchi, K., Matsuoka, N., Tsuboi,M., Koji, T., Urayama, S., Fujiyama, K., Kii-iyaina,T., Nakashima, T., Nakane, P. K. and Negataki, S.Fas and Fas ligand interaction is necessary for humanosteoblast apoptosis. J. Bone Miner. Res. 1997, 12,1637-1646
  17. Kerr, J. B. Spontaneous degeneHtion of germ cells inrat testis: Assessrnent of cell types and frequencyduring the spennatogenic cycle. J. Reprod. Rsrdl. 1992,95, 825-830
  18. Koji, T., Hishikawa, Y., Ando, H., Nakanishi, Y.and Kobaya-shi, N. Expression testes: mvolvement ofthe Fas system in the induction of germ cell apoptosisin the damaged mouse testis. Biol. Reprod. 2001, 64,946-952
  19. Koji, T., Ohta, H., Kobayashi, N., Nakanishi, Y.and Nakne, P. K. Involvement of Fas/Fas ligandsystem in the induction of germcell apoptosis innormal and ischemia-reperfusion testis of adult mouse.Cell Struct, Funct. 1997, 22, 757-761
  20. Kothari, L. K., Mishra, P. and Mishra, R. K. Effectof bilateral vasectomy on the structure and function ofthe testes. Am. J. Surg. 1973, 126, 84-88
  21. Kubota, Y., Sasaki, S., Kubota, HL, Tatsura, H. andKohri, K. A study on the mechanism of thespermatogenic damage after vasectomy in rats. NipponHinyotdka Gakkai Zasshi. 2001, 92, 13-22
  22. Lee, J., Richburg, J. H., ShipP, E. B., Meistnch, M.L. and Boekelheide, K. The Pas system, a regulatorof testicular germ cell apoptosis, is differentiallyup-regulated in Sertoli cell versus germ cell injury ofthe testis. Endocrin. 1999, 140, 852-858
  23. Lee, J., Richburg, J. H., Younkin, S. C. andBoekelheide, K. Fas system is a key regulator ofgerm cell apoptosis in the testis. Endocrin. 1997, 138,2081-2088
  24. Lue, Y., Sinha Hikim, A. P., Wang, C., Bonavera,J. J., Baravarian, S., Leung, A. and Swerdloff, R.S. Early effect of vasectomy on testicular structure and on germ cell and macrophage apoptosis in the hamster.J. Androl. 1997, 18, 166-173
  25. McClure, R. F., Heppelmann, C. J. and Paya, C. V.Constitutive Fas ligand gene transcription in Sertolicells is regulated by Spl. J. Biol. Chem. 1999, 274,7756-7762
  26. Nagata, S. and Golstein, P. The Fas death factor.Science. 1995, 267, 1449-1456
  27. Nandi, S., Banerjee, P. P. and Zirkin, B. R. Germcell apoptosis in the testes of Spraque (-) Dawley ralsfollowing testasteixxie withdrawal by ethane 1,2-dimethanesulfonate administration: relationship to Fas? Biol.Reprod. 1999, 61, 70-75
  28. Nogae, S., Miyazaki, M., Kobayashi, N., Saito, T.,Abe, K., Saito, KL,Nakane, P. K., Nakanishi, Y. andKoji, T. Induction of apoptosis in ischemia-reperfusionmodel of mouse kidney: possible involvement of FasJ. Am. Soc. NephroL 1998, 9, 620-631
  29. Nonclercq, D., Reverse, D., Toubeau, G., Beckers, J.F., Sulon, J. L., Zanen, J. and Heuson-Stiennon, J.A. In situ demcnstration of germinal oell apoptosis duringdiethylstilbestrol-mduced testis regression in adult maleSyrian hamsters. Biol. Reprod. 1996, 55, 1368-1376
  30. Okada, K., Komuta, K., Hashimoto, S., Matsuzaki,S., Kanematsu, T. and Koji, T. Frequency ofapoptosis of tumor-infiltrating lymphocytes induced byFas counterattack in human colorectal carcinoma andits correlation with prognosis. Clin. Cancer Res. 2000,6, 3560-3564
  31. Pentikainen, V., Erkkila, K. and Dunkel, L. Fasregulates germ cell apoptosis in the human testis invitro. Am. J. Physiol. 1999, 276, E310-E316
  32. Richburg, J. H., Nanez, A. and Gao, H. Participationof the Fas-signaling system in the initiation of germcell apoptosis in young rat testis after exposure tomono-(2-ethy1hexy1) Phthalate. Toxicol. Appl. Phanracol.1999, 160, 271-278
  33. Richburg, J. H., Nanez, A-, Williams, L. R., Embree,M. E. and Boekelheide, K. Sensitivity of testiculargerm cells to toxicant-induced apoptosis in gld micethat express a non-functional form of Fas ligand.Endocrin. 2000, 141, 787-793
  34. Rodriguez, L, Ody, C., Araki, K., Garcia, I. andVassalli, P. An early and massive wave of germinalcell apoptosis is required for the development offuncdonal spermatogenesis. EMBO J. 1997, 16, 2262-2270
  35. Russell, L. D., Ettlin, R. A., Sinha Hikim, A. P.and Clegg, E. D. Staging for laboratory species. InHistological and Histopathological Evaluation of theTesds, PP. 62-118, Cache River Pnss, Qearwater, 1990
  36. Russell, L. D. and Clennont, Y. Degeneration ofgerm cells in normal, hypophysectomized and hormonetreated hypophysectomized rats. Anat. Rec. 1997, 187,347-366
  37. Sarrat, R., Whyte, J., Torres, A., Lostale, F. andDiaz, M. P. Experimental vasectomy and tesdcularstructure. Histol. Histopathol. 1996, 11, 1-6
  38. Shiraishi, K., Naito, K. and Yoshida, K. Vasectomyimpairs spermatogenesis through germ cell apoptosismediated by p53-Bax pathway in rat. J. Urol. 2001,166, 1565-1571
  39. Shiraishi, K., Yoshida, K., Fujimiya, T. and Naito,K. Activation of mitogen activated protein Idnases andapoptosis of germ cells after vasectomy in the rat. J.Urol. 2002, 168, 1273-1278
  40. Sinha Hikim, A. P., Rajavashisth, T. B., SinhaHikiin, I., Lue, Y., Bonavera, J. J., Leung, A.,Wang, C. and Swerdloff, R. S. Significance ofapoptosis in the temporal and stage-specific loss ofgerm cells in the adult rat after gonadotropindeprivation. Biol. Reprod. 1997, 57, 1193-1197
  41. Stephan, H, Polzar, B., Rauch, F., Zanotti, S., Ulke,C. and Mannherz, H. G. Distribution of deoxyri-bonuclease I (DNase I) and p53 in rat testis and theircorrelation with apoptosis. Histochem. Cell Biol. 1996,106, 383-393
  42. Suda, T., Takahashi, T., Golstein, P. and Nagata, S.Molecular cloning and expression of the Fas ligand, anovel member of the tumor necrosis factor family.Cell. 1993, 75, 1169-1178
  43. Tamura, M., Kimura, H., Koji, T., Tominaga, T.,/Vshizav'a, K., Kiriyama, T., Yokqyama, N., Yoshirmra,T., Eguchi, K., Nakane, P. K. and Nagataki, S. Roleof apoptosis of thymocytes in a rat model of goiter. Apossible involvement of Fas system. Endocrin. 1998,139, 3646-3653
  44. Yin, Y., DeWoIf, W. and Morgentaler, A. Experinienlalcrytorchidism induces testicular germ cell apoptosis byp53-dependent and -independent pathway in mice.Biol. Reprod. 1998, 58, 492-496