Intraoperative evaluation of testicular vascularization and perfusion in rat testicles with indocyanine green (ICG)/near-infrared (NIR) fluorescent imaging after torsion-detorsion and reperfusion.

INTRODUCTION: The aim of this study was to evaluate testicular perfusion and vascularization with intraoperative ICG/NIR imaging in a testicular ischemia-reperfusion model and to investigate the effects of ICG on testicular tissue. MATERIALS AND METHODS: 24 male rats were divided into four groups. In the ICG group, only ICG was given and images of the testicles were recorded with NIR camera. In the torsion group, the testicles were left in torsion for 4 h. ICG/NIR images were obtained after torsion and detorsion. In the reperfusion group, ICG/NIR images of the testicles were obtained at the 4th hour of reperfusion. After the procedures, testicles were collected and evaluated with histological, immunohistochemical examination and qRT-PCR. RESULTS: There was no histologically negative effect of ICG on testicular tissue. There was no testicular perfusion in the torsion group, but perfusion started after detorsion. At the 4th hour of reperfusion, testicular perfusion continued. TNF-a, IL-6, MCP-1 and caspase-3 immunoreactivity were found to be at low levels in the control and ICG groups, while high in the torsion and reperfusion groups (p < 0.05). In qRT-PCR, TNF-a, IL-6, MCP-1 and caspase-3 expressions were lower in the control and ICG groups, but higher in the torsion and reperfusion groups. CONCLUSION: There was no histologically negative effect of ICG on testicles. The ICG/NIR imaging technique seems to be a feasible method in testicular torsion and may contribute to the surgeon in the intraoperative management of testicular torsion. In testicles that started to be perfused after detorsion, perfusion still continued at the 4th hour of reperfusion. Our next goal is to test whether testicles showing ICG fluorescence in during reperfusion maintain their viability for long term.

Effect of High dose Gonadotropin Stimulation on Follicular Atresia through Light Chain 3B and Voltage Dependent Anion Channel 2.

Background: Follicle development takes place under the control of hormonal and environmental stimuli. It suggested that to improve in vitro fertilisation outcomes in poor responders increasing gonadotropin doses be used. Excessive gonadotropin leads to atresia and impairs follicular development, but the molecular mechanisms of follicular atresia remain largely unknown. Recently, it was suggested that autophagy may be an alternative mechanism involved in follicle depletion. Aims: In this study, we aimed to clarify the role of autophagic markers such as light chain (LC) 3B and voltage dependent anion channel 2 (VDAC2) in follicular atresia using the high dose gonadotropin stimulation. Settings and Design: The female 24 BALB/c mice were employed in the present study under the Committee for the Purpose of Control and Supervision of Experiments on Animals guidelines with ethical clearance from the institutional ethical committee. These mice were categorised into four groups, with six rats in each as control and test animals. Materials and Methods: Group 1 (control): no action will be taken. Group 2 (sham): only saline will be applied. Group 3: low-dose gonadotropin Pregnant mare's serum gonadotropin (PMSG) + human chorionic gonadotropin (HCG) will be applied. Group 4: high-dose gonadotropin + HCG will be applied. The animals were sacrificed 48 h after the last injection. For all group samples, both protein and mRNAs of the LC3B and VDAC2 were examined by immunohistochemical and reverse transcription-polymerase chain reaction techniques. Statistical Analysis Used: All variables were analysed using GraphPad Prism 8. Kruskal-Wallis t-test and Mann-Whitney U test were used to compare immunohistochemical results; in addition to this, parametric one-way ANOVA test and Shapiro-Wilk test were applied for quantitative polymerase chain reaction statistics. Results: An increased number of atretic follicles were observed in the high-dose gonadotropin + HCG group. LC3B immunoreactivity of the atretic secondary follicles in the high-dose group is higher than in other groups. The expression of VDAC2 protein in the secondary and Graafian follicles and also VDAC2 mRNA in the ovary were more highly expressed in the control and sham groups. The decrease in VDAC2 mRNA level and immunohistochemical expression was remarkable in the low-dose and high-dose follicle-stimulating hormone groups compared to the control and sham groups. Conclusion: In this study, the increased LC3B and decreased VDAC2 expression, which are autophagy markers, were observed in both the gonadotropins groups, so we suggested that high doses of gonadotropins may cause ovarian atresia.

Cyclophosphamide suppresses spermatogenesis in the testis of mice through downregulation of miR-34b and miR-34c.

Cyclophosphamide (CP) is commonly used as an anticancer agent but has been associated with high toxicity in several organs, including the testes. In this study, we aimed to evaluate the effects of CP-induced testicular toxicity, using glial cell line-derived neurotrophic factor (GDNF), occludin and transforming growth factor beta 3 (TGF-ß3) primary antibodies, and miR-34b and miR-34c expressions. Eighteen young Balb/c male mice were divided into three groups. The control group received no treatment. The mice of CP group were injected 100 mg kg-1  day-1 CP for 5 days, and the same amount of saline was injected in the sham group. The animals were sacrificed 24 hr after the last injection. Immunohistochemical analysis of testicular tissues showed a decrease in both spermatogenic germ cell count and also GDNF, occludin expressions, but an increase in TGF-ß3 expression in the CP group compared to the others group. The expressions of miR-34b and miR-34c were examined by qPCR technique, a significant decrease was observed in tissue samples in the CP-treated group. The expression of GDNF, occludin and TGF-ß3 plays an important role in testicular injury caused by CP, and the decrease in the expression of miR-34b/c in tissue samples may be an important marker for the detection of testicular damage.