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MP43-19 RELATIONSHIP BETWEEN SERTOLI CELL FUNCTION AND DIFFERENTIATION PROCESS OF SPERMATOGONIAL STEM CELLS IN CRYPTORCHID TESTES
THE JOURNAL OF UROLOGYâ
e586
Vol. 195, No. 4S, Supplement, Sunday, May 8, 2016
RESULTS: Between 2009 and 2015, 46 patients underwent microscopic varicocele repair. Ten patients met the inclusion criteria. Mean age at time of surgery¼ 16.3 years (15-18 yrs). Mean TVDiff ¼ 5.9% (0-16%). Mean duration of postoperative SA collection¼ 36 weeks (17-40 weeks). As noted in the table, the difference in median TMSC before and after surgery was highly statistically significant (p¼ 0.0039). CONCLUSIONS: Varicocele repair significantly improves TMSC in adolescent, Tanner V stage males presenting with asymptomatic varicoceles. In view of this pilot data, a larger multi-institutional prospective study comparing various treatment modalities for varicoceles is warranted.
Source of Funding: Work supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number U54-GM104941 (PI: Binder-Macleod).
Source of Funding: none
MP43-20 TESTICULAR HYPOPLASIA IS DRIVEN BY DEFECTIVE VASCULAR FORMATION
MP43-19 RELATIONSHIP BETWEEN SERTOLI CELL FUNCTION AND DIFFERENTIATION PROCESS OF SPERMATOGONIAL STEM CELLS IN CRYPTORCHID TESTES Takashi Nagai*, Kentaro Mizuno, Yutaro Hayashi, Hideyuki Kamisawa, Yoshinobu Moritoki, Hidenori Nishio, Satoshi Kurokawa, Akihiro Nakane, Tetsuji Maruyama, Takahiro Yasui, Nagoya, Japan INTRODUCTION AND OBJECTIVES: Spermatogonial stem cells (SSCs) in the testes serve as a source of spermatozoa after puberty, and are differentiated from gonocytes during 0 mini-puberty,0 immediately after birth. By using a cryptorchid animal model, we found that the epigenetic regulation of certain genes influences the SSC differentiation process; however, the entire differentiation process is not completely clear. Some recent studies propose the importance of the microenvironment surrounding the SSCs, including Sertoli cells, in this process. To clarify the role of Sertoli cells, we examined the relationship between the number of SSCs and hormonal regulation associated with Sertoli cells. METHODS: Of 93 prepubertal patients with cryptorchidism who underwent orchiopexy from January 2013 to July 2015 in our hospital, we enrolled 38 patients. The mean age at surgery was 23.9 (range, 7 137) months. The serum LH, FSH, testosterone, anti-Mullerian hormone (AMH), and Inhibin-B (INHBB) levels were measured preoperatively, and testicular biopsy was performed intraoperatively. The number of SSCs were counted in 50 randomly selected seminiferous tubules through immunohistochemical staining for DEAD box protein 4 - an SSC-specific marker. This study was approved by the institutional review board (#83). RESULTS: The serum AMH and INHBB levels were 198.5 (range, 46.8 - 414.0) ng/ml and 120.3 (range, 98.0 - 142.0) pg/ml, respectively; these values were not correlated with age at surgery. The number of SSCs per seminiferous tubule was 1.19 (range, 0.3 - 2.38). The number of SSCs was not significantly correlated with the serum LH, FSH, testosterone, and INHBB levels, but was inversely correlated with the serum AMH levels (r¼-0.368, p<0.05, Graph). CONCLUSIONS: Both AMH and INHBB were secreted from Sertoli cells following stimulation by FSH; these hormones are considered to reflect the capability of Sertoli cells after birth. In male embryos, AMH triggers the degeneration of the uterine precursor; however, the testes continue to secrete AMH throughout development and into adulthood. We noted that the serum AMH levels increased in patients with a decreased number of SSCs. Hence, we believe that dysfunction of Sertoli cells may affect the differentiation process from gonocytes to SSCs.
udia Salgado, Miguel Reyes-Mugica, Pankaj Dangle*, Dr. Cla Rajeev Chaudhry, Francis Schneck, Michael Ost, Sunder Sims-lucas, Pittsburgh, PA INTRODUCTION AND OBJECTIVES: A testicular “nubbin” or vanishing testis is considered to be secondary to a neo-natal torsion event and is evidenced by a definable hemosiderin deposit. We hypothesize that a, “vanishing testis” is a fault in embryological development as a result of arrest in endothelial cells migration rather than secondary to a random physical torsion/twist. Endothelial cell migration and testicular cord formation are interdependent processes, involving rearrangement of migrating endothelial cells from the mesonephric vascular plexus. We therefore sought to determine whether the testicular microvasculature is underdeveloped in “nubbin” testis compared with age matched healthy testicles from cadaveric specimens. METHODS: Cases of testicular nubbin excision [average age 9.5 (range 2-12) months] were compared with age-matched controls [5.14, (2- 12 months)] from cadaveric testes without known genitourinary pathology. Testicular samples were serially sectioned and sampled appropriately at selected levels for vascular labeling. To elucidate the testis microvasculature we performed immunohistochemistry (IHC) using an automated staining platform with controlled and standardized conditions and positive and negative controls. We used CD34 which in the testis stains the endothelium, stem cells and interstitium; CD31 to stain the endothelium; and D2-40 to stain lymphatic endothelium. Whole slide digital images were subsequently scanned and morphometric analysis was carried out, the percentage of the total tissue with CD31 and CD34 positive stain was assessed and the number of the lymphatic vessels (D2-40) per mm2 was counted. RESULTS: Seven patients were found to have vanishing testis on the left side. Of the 10 cases stained, 4 had evidence of hemosiderin deposit and calcification. The average testicular volume in these 4 cases was 2.2 compared to 4.8 cm3 in others with diagnosis of testicular atrophy (p¼0.103). The percentage (%) distribution of CD34 in controls was higher, 14.44.5 (mean SD) compared to nubbin cases 7.53.8 (p¼0.002). The % distribution of CD31 was 3.21.7 in controls and trended to be less in nubbin testis (2.21.0) (p¼0.152). The lymphatic distribution was unchanged in both case and control. These findings elucidate that the testis microvasculature is mis-patterned in nubbin cases, while the lymphatic endothelium distribution is unaffected. CONCLUSIONS: This histopathological study suggests that disturbances in endothelial malformation may be a contributing factor leading to testicular hypoplasia and a resultant nubbin testis independent of a physical torsion event. Source of Funding: none
e586
Vol. 195, No. 4S, Supplement, Sunday, May 8, 2016
RESULTS: Between 2009 and 2015, 46 patients underwent microscopic varicocele repair. Ten patients met the inclusion criteria. Mean age at time of surgery¼ 16.3 years (15-18 yrs). Mean TVDiff ¼ 5.9% (0-16%). Mean duration of postoperative SA collection¼ 36 weeks (17-40 weeks). As noted in the table, the difference in median TMSC before and after surgery was highly statistically significant (p¼ 0.0039). CONCLUSIONS: Varicocele repair significantly improves TMSC in adolescent, Tanner V stage males presenting with asymptomatic varicoceles. In view of this pilot data, a larger multi-institutional prospective study comparing various treatment modalities for varicoceles is warranted.
Source of Funding: Work supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number U54-GM104941 (PI: Binder-Macleod).
Source of Funding: none
MP43-20 TESTICULAR HYPOPLASIA IS DRIVEN BY DEFECTIVE VASCULAR FORMATION
MP43-19 RELATIONSHIP BETWEEN SERTOLI CELL FUNCTION AND DIFFERENTIATION PROCESS OF SPERMATOGONIAL STEM CELLS IN CRYPTORCHID TESTES Takashi Nagai*, Kentaro Mizuno, Yutaro Hayashi, Hideyuki Kamisawa, Yoshinobu Moritoki, Hidenori Nishio, Satoshi Kurokawa, Akihiro Nakane, Tetsuji Maruyama, Takahiro Yasui, Nagoya, Japan INTRODUCTION AND OBJECTIVES: Spermatogonial stem cells (SSCs) in the testes serve as a source of spermatozoa after puberty, and are differentiated from gonocytes during 0 mini-puberty,0 immediately after birth. By using a cryptorchid animal model, we found that the epigenetic regulation of certain genes influences the SSC differentiation process; however, the entire differentiation process is not completely clear. Some recent studies propose the importance of the microenvironment surrounding the SSCs, including Sertoli cells, in this process. To clarify the role of Sertoli cells, we examined the relationship between the number of SSCs and hormonal regulation associated with Sertoli cells. METHODS: Of 93 prepubertal patients with cryptorchidism who underwent orchiopexy from January 2013 to July 2015 in our hospital, we enrolled 38 patients. The mean age at surgery was 23.9 (range, 7 137) months. The serum LH, FSH, testosterone, anti-Mullerian hormone (AMH), and Inhibin-B (INHBB) levels were measured preoperatively, and testicular biopsy was performed intraoperatively. The number of SSCs were counted in 50 randomly selected seminiferous tubules through immunohistochemical staining for DEAD box protein 4 - an SSC-specific marker. This study was approved by the institutional review board (#83). RESULTS: The serum AMH and INHBB levels were 198.5 (range, 46.8 - 414.0) ng/ml and 120.3 (range, 98.0 - 142.0) pg/ml, respectively; these values were not correlated with age at surgery. The number of SSCs per seminiferous tubule was 1.19 (range, 0.3 - 2.38). The number of SSCs was not significantly correlated with the serum LH, FSH, testosterone, and INHBB levels, but was inversely correlated with the serum AMH levels (r¼-0.368, p<0.05, Graph). CONCLUSIONS: Both AMH and INHBB were secreted from Sertoli cells following stimulation by FSH; these hormones are considered to reflect the capability of Sertoli cells after birth. In male embryos, AMH triggers the degeneration of the uterine precursor; however, the testes continue to secrete AMH throughout development and into adulthood. We noted that the serum AMH levels increased in patients with a decreased number of SSCs. Hence, we believe that dysfunction of Sertoli cells may affect the differentiation process from gonocytes to SSCs.
udia Salgado, Miguel Reyes-Mugica, Pankaj Dangle*, Dr. Cla Rajeev Chaudhry, Francis Schneck, Michael Ost, Sunder Sims-lucas, Pittsburgh, PA INTRODUCTION AND OBJECTIVES: A testicular “nubbin” or vanishing testis is considered to be secondary to a neo-natal torsion event and is evidenced by a definable hemosiderin deposit. We hypothesize that a, “vanishing testis” is a fault in embryological development as a result of arrest in endothelial cells migration rather than secondary to a random physical torsion/twist. Endothelial cell migration and testicular cord formation are interdependent processes, involving rearrangement of migrating endothelial cells from the mesonephric vascular plexus. We therefore sought to determine whether the testicular microvasculature is underdeveloped in “nubbin” testis compared with age matched healthy testicles from cadaveric specimens. METHODS: Cases of testicular nubbin excision [average age 9.5 (range 2-12) months] were compared with age-matched controls [5.14, (2- 12 months)] from cadaveric testes without known genitourinary pathology. Testicular samples were serially sectioned and sampled appropriately at selected levels for vascular labeling. To elucidate the testis microvasculature we performed immunohistochemistry (IHC) using an automated staining platform with controlled and standardized conditions and positive and negative controls. We used CD34 which in the testis stains the endothelium, stem cells and interstitium; CD31 to stain the endothelium; and D2-40 to stain lymphatic endothelium. Whole slide digital images were subsequently scanned and morphometric analysis was carried out, the percentage of the total tissue with CD31 and CD34 positive stain was assessed and the number of the lymphatic vessels (D2-40) per mm2 was counted. RESULTS: Seven patients were found to have vanishing testis on the left side. Of the 10 cases stained, 4 had evidence of hemosiderin deposit and calcification. The average testicular volume in these 4 cases was 2.2 compared to 4.8 cm3 in others with diagnosis of testicular atrophy (p¼0.103). The percentage (%) distribution of CD34 in controls was higher, 14.44.5 (mean SD) compared to nubbin cases 7.53.8 (p¼0.002). The % distribution of CD31 was 3.21.7 in controls and trended to be less in nubbin testis (2.21.0) (p¼0.152). The lymphatic distribution was unchanged in both case and control. These findings elucidate that the testis microvasculature is mis-patterned in nubbin cases, while the lymphatic endothelium distribution is unaffected. CONCLUSIONS: This histopathological study suggests that disturbances in endothelial malformation may be a contributing factor leading to testicular hypoplasia and a resultant nubbin testis independent of a physical torsion event. Source of Funding: none