The Fas system may have a role in male reproduction

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The Fas system may have a role in male reproduction Ciler Celik-Ozenci, D.D.S., Ph.D., Zeliha Sahin, M.Sc., Ismail Ustunel, Ph.D., Gokhan Akkoyunlu, Ph.D., Tibet Erdogru, M.D., Emin Turkay Korgun, Ph.D., Mehmet Baykara, M.D., Ramazan Demir, Ph.D.  Fertility and Sterility  Volume 85, Pages 1168-1178 (April 2006) DOI: 10.1016/j.fertnstert.2005.08.058 Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Localization of FasL protein in sham-operated (a–c) and varicocele-induced (d–h) rat testis. (a) FasL first appears in Golgi vesicles of round spermatids (arrowheads and inset), (b) then in acrosomal caps of round spermatids (arrowheads and inset), and (c) finally in heads of elongating spermatids (arrowheads and inset). (d) In the 9-week-old varicocele group, the expression of FasL was similar to that of control, as each phase of the expression pattern (arrows) was seen in tubuli sections. (e, f) In the 11-week-old varicocele group, there were cells still expressing FasL in their heads (arrows), whereas some cells did not express this protein accurately (arrowheads and insets). (g, h) In the 13-week-old varicocele group, some cells were negative in the same tubule (arrowheads), whereas some cells were positive for FasL (arrows). (i) Negative-control section; note the absence of FasL immunostaining. Scale bars represent 100 μm (panel i) or 10μm (panels a–h). Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Localization of Fas protein in sham-operated (a) and varicocele-induced (b–d) rat testis. (a) Fas was present in cytoplasmic extrusions of elongated spermatids (arrow) and also in Leydig cells of the interstitial tissue (inset, arrowhead). (b) Similar to control, Fas was present in cytoplasmic extrusions (arrow) and in Leydig cells (arrowhead and inset) in the 9-week-old varicocele group. (c) Diminished intensity for Fas in Leydig cells can be seen (arrowhead and inset) in the 11-week-old varicocele group. (d) There were still a few cells immunostained with Fas in the 13-week-old varicocele group (arrowhead and inset). Scale bars represent 10 μm. Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 Detection of TUNEL-positive apoptotic germ cell nuclei (arrows) in seminiferous tubular cross section. (a) Sham, (b) 9-week-old varicocele, (c) 11-week-old varicocele, and (>d) 13-week-old varicocele. Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 4 Expression of Fas and FasL proteins in sham and different varicocele weeks determined by Western blot analysis using β-actin as an internal standard. Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 5 Quantitative analysis of Western blot results (per-group n = 6). The values in the bar graphs were obtained by dividing the protein intensity value to its corresponding β-actin intensity value. (a) Fas protein was significantly reduced in the 13-week-old varicocele group when compared with sham. (b) FasL expression was significantly less in the 11-week and 13-week-old varicocele groups when compared with sham. *P<.05. Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions

FIGURE 6 Ultrastructural features of varicocele-induced rat testicular tissue. (a) General appearance of germinal epithelium in the seminiferous tubule wall of the 13-week-old varicocele group. (b) Arrows indicate the disturbance of contact points between spermatogonium (SG) and Sertoli cell (ST) in the 13-week-old varicocele group. L = lumen. Scale bars represent 10 μm (a) and 2 μm (b). Celik-Ozenci. Fas system in varicocele-induced rat testis. Fertil Steril 2006. Fertility and Sterility 2006 85, 1168-1178DOI: (10.1016/j.fertnstert.2005.08.058) Copyright © 2006 American Society for Reproductive Medicine Terms and Conditions