Effect of experimental varicocele on the expressions of notch 1, 2, and 3 in rat testes: An immunohistochemical study  Zeliha Sahin, M.Sc., Zubeyde Bayram, M.Sc., Ciler Celik-Ozenci, Ph.D., Gokhan Akkoyunlu, Ph.D., Yasemin Seval, M.Sc., Tibet Erdogru, M.D., Ismail Ustunel, Ph.D., Mehmet Baykara, M.D., Ramazan Demir, Ph.D.  Fertility and Sterility  Volume 83, Issue 1, Pages 86-94 (January 2005) DOI: 10.1016/j.fertnstert.2004.09.006 Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 1 Hematoxylin and eosin stained rat testicular tissues. When compared with (A) the control sham-operation rats, (B) 9-week varicocele rats, (C) 11-week varicocele rats, and (D) 13-week varicocele rats showed an apparent degeneration of the tubules that was related to the progressing weeks of varicocele (magnification ×50). (E) The percentage of the calculated degenerated tubules in varicocele groups. A statistically significant (P<.05) increase was found in the degenerating tubule percentage of 11-week and 13-week varicocele groups when compared with the controls (one way ANOVA followed by post hoc Tukey). Pink bar: Sham operation. Orange bar: 9-week varicocele. Red bar: 11-week varicocele. Blue bar: 13-week varicocele. *P<.05. Fertility and Sterility 2005 83, 86-94DOI: (10.1016/j.fertnstert.2004.09.006) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 2 Immunohistochemical detection of Notch 1 (A–E), Notch 2 (F–K), and Notch 3 (L–P) receptors in testis tissue of rats. Notch-1 receptor immunoreactivity: (A) in sham-operated rats, where arrows indicate sperm cells (insert: Leydig cells); (B) in 9-week varicocele rats, the staining was similar to the sham-operated rats (insert: Leydig cells); (C) in 11-week varicocele rats, the staining was limited to Leydig cells; (D) in 13-week varicocele rats, no positive immunostaining is seen. Notch-2 receptor immunoreactivity: (F) in sham-operated rats, arrows indicate Leydig cells; (G) in sham-operated rats, arrows indicate primary spermatocytes; (H) in 9-week varicocele rats, Leydig cells (insert) are similar to the sham-operated group whereas staining of the other cells decreased; (I) in 11-week varicocele rats, staining decreased in all cells including Leydig cells (insert); (J) in 13-week varicocele rats, the staining decreased in Leydig cells, spermatogonium, and primary spermatocytes when compared with the sham-operation group. Notch-3 receptor immunoreactivity: (L) in sham-operated rats, (M) in 9-week varicocele rats, and (N) in 11-week varicocele rats, weak immunoreactivity was present in Leydig cells (inserts); however, (O) in 13-week varicocele rats, no immunoreactivity was found. (E, K, P) Negative control sections. Magnification ×100. Fertility and Sterility 2005 83, 86-94DOI: (10.1016/j.fertnstert.2004.09.006) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions

FIGURE 3 HSCORE for (A) Notch 1, (B) Notch 2, and (C) Notch 3 receptors in sham-operation and varicocele-induced groups. A statistically significant decrease was found in the expression of Notch 1 (A) receptor in 11-week and 13 week varicocele groups as well as a statistically significant decrease in the expression of Notch 2 (B) and Notch 3 (C) receptors in 9-week, 11-week, and 13-week varicocele groups when compared with the control group (Mann-Whitney U test). Pink bar: Sham-operation; Orange bar: 9-week varicocele; Red bar: 11-week varicocele; Blue bar: 13-week varicocele. (*P<.05). Fertility and Sterility 2005 83, 86-94DOI: (10.1016/j.fertnstert.2004.09.006) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions