Testicular localization of activating transcription factor 1 and its potential function during spermatogenesis†.

Activating transcription factor 1 (ATF1), belonging to the CREB/ATF family of transcription factors, is highly expressed in the testes. However, its role in spermatogenesis has not yet been established. Here, we aimed to elucidate the impact of ATF1 in spermatogenesis by examining the expression pattern of ATF1 in mice and the effect of ATF1 knockdown in the mouse testes. We found that ATF1 is expressed in various organs, with very high levels in the testes. Immunohistochemical staining showed that ATF1 was localized in the nuclei of spermatogonia and co-localized with proliferating cell nuclear antigen. In ATF1-deficient mice, the seminiferous tubules of the testis contained cells at all developmental stages; however, the number of spermatocytes was decreased. Proliferating cell nuclear antigen expression was decreased and apoptotic cells were rare in the seminiferous tubules. These results indicate that ATF1 plays a role in male germ cell proliferation and sperm production.

Transcriptome Analysis to Identify Human Spermatogonial Cells from Sertoli Cell-Only Testes.

PURPOSE: We sought to determine whether gene products from spermatogenic cells could be detected in Sertoli cell-only testes. Transcriptome analysis using next generation sequencing was performed using human testicular samples. MATERIALS AND METHODS: This study enrolled 20 men with nonobstructive azoospermia with a histological diagnosis of Sertoli cell-only and no germ cells on in vitro fertilization analysis and 5 with obstructive azoospermia and histologically normal spermatogenesis. Individual differences in the transcriptome of obstructive azoospermia testicular tissues generated on the Illumina® platform were compared as the number of fragments per kilobase of exon per million mapped sequence reads. We confirmed mRNA expression and targeted gene product localization by quantitative reverse transcriptase-polymerase chain reaction and immunohistochemical analysis, respectively. RESULTS: Using next generation sequencing we analyzed 5,666 of the 23,003 screened genes. As representative markers of immature germ cells, UTF1, CD9, DDX4, EPCAM, GFRA1, KIT, LIN28, DMRT, GPR125, UCHL1 and NANOG transcripts were detected in 13 Sertoli cell-only samples (65%). Reverse transcriptase-polymerase chain reaction showed significant mRNA expression of UTF1, CD9, DDX4 and KIT in Sertoli cell-only samples. Immunostaining revealed the localization of DDX4-positive cells, presumably spermatogonia, in 9 Sertoli cell-only samples (45%). These cells lacked proliferative cell nuclear antigen expression. CONCLUSIONS: A number of Sertoli cell-only testes contain immature germ cells (spermatogonia and spermatogenic stem cells) with various gene expression profiles. Sertoli cell-only is a heterogeneous pathophysiology which may be corrected by medical treatment or regenerative technologies.

High Inguinal Microsurgical Denervation of the Spermatic Cord for Chronic Scrotal Content Pain: A Novel Approach for Adult and Pediatric Patients.

OBJECTIVE: To improve the technique and results of microsurgical denervation of the spermatic cord (MDSC) for men with chronic scrotal content pain, we describe a novel approach at the level of the internal inguinal ring for the complete transection of the nerves running both inside and outside the spermatic cord for adults and children. METHODS: A retrospective review of 52 patients (64 testicular units) who underwent high inguinal MDSC was performed. Visual analogue scale (VAS, 1-10) scores were compared with before and every 3 months after the surgery. Depressive symptoms were assessed by the Beck Depression Inventory. Hormonal evaluations were performed before and 6 months after the surgery. RESULTS: The average patient age was 52.4 years (12-78); including 6 pediatric cases. The mean operative time was 67 minutes per testicular unit, and there were no major complications. The mean pre- and post-MDSC VAS scores were 8.3 and 2.5, respectively (P < .0001). Forty-six (88%) cases showed positive responses after MDSC, and multivariate analysis showed that pain outside the scrotum and depressive symptoms were predictors of MDSC failure (P < .05, odds ratio: 15.27 and 12.56, respectively). CONCLUSION: For both adult and pediatric patients, high inguinal MDSC is an effective and safe management option, including testicular function, for the chronic scrotal content pain that is refractory to medical management. We find that the high inguinal approach is easier in our experience than the subinguinal approach because of fewer divisions of veins, a larger diameter of the spermatic artery.