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Recent evidence suggests that neutrophil recruitment may initiate germ cell apoptosis in the ischemic testis. The purpose of the present study was to examine the relationship between germ cell apoptosis and neutrophil recruitment in the contralateral testis following testicular ischemia-reperfusion (IR) injury in a rat. Adult male Sprague-Dawley rats were divided randomly into two experimental groups: Group A: Sham operated animals; Group B: IR rats underwent 90 min of unilateral testicular ischemia following by 96 h of reperfusion. The rats were sacrificed and testes were harvested. Johnsen's criteria and the number of germinal cell layers were measured to categorize the spermatogenesis. TUNEL assay was used to determine germ cell apoptosis in both the ischemic and contralateral testis. The recruitment of neutrophils was calculated per 100 venules. Expression of E-selectin was determined using immunohistochemical analysis. Statistical analysis was performed using Student's t test, with P less than 0.05 considered statistically significant. Germ cell apoptosis in both the ischemic and the contralateral testis increased significantly after IR. E-selectin expression was significantly greater in ischemic testis from IR rats compared to sham animals. The small increase in E-selectin expression and the concomitant increase in neutrophil recruitment in the contralateral testis of the IR rats (vs. sham animals) were not statistically significant. In conclusion, testicular ischemia causes an increase in germ cell apoptosis in the contralateral testis. Mechanisms other than neutrophil recruitment apparently initiate this process.

Objective: To examine the relationship between the time of reperfusion and neutrophil recruitment, E-selectin expression, and germ cell apoptosis in the ischemic and contralateral testis after testicular ischemia-reperfusion (IR) injury in a rat. Design: Laboratory study. Setting: Research laboratory in a faculty of medicine at Technion–institute of technology in Israel. Animal(s): Sixty adult Sprague-Dawley rats weighing 250–280 g. Intervention(s): Testicular IR. Main Outcome Measure(s): Testicular germ cell apoptosis was assessed by terminal deoxynucleotide transferase–mediated 2′-deoxyuridine 5′-triphosphate nick-end labeling immunohistochemical assay, using an in situ cell death detection kit. The recruitment of polymorphonuclear (PMN) cells was calculated per 100 venules. Expression of E-selectin was determined by using immunohistochemical analysis. Result(s): E-selectin expression and polymorphonuclear-cell recruitment in the contralateral testis increased significantly after 1 hour of reperfusion and then remained unchanged during the first 24–48 hours, followed by a gradual decrease. Germ cell apoptosis in the contralateral testis increased after 6 hours of reperfusion, achieved statistical significance after 24 hours, and decreased after 72 hours of reperfusion. Conclusion(s): Germ cell apoptosis in the contralateral testis increases most significantly within the first 24–48 hours, followed by a gradual decrease, after IR injury. E-selectin expression and neutrophil recruitment increases within the first 6 hours and apparently may initiate the increase in germ cell apoptosis. [Copyright &y& Elsevier]