Changes in Inflammatory Cytokines Accompany Deregulation of Claudin-11, Resulting in Inter-Sertoli Tight Junctions in Varicocele Rat Testes

Oh, Y.S.,Jo, N.H.,Park, J.K.,Gye, M.C. Williams and Wilkins Co 2016 The Journal of urology Vol.196 No.4

<P>Purpose: To elucidate the changes that occur in the blood-testis barrier during varicocele we examined changes in Cldn11 (claudin-11), an element of the blood-testis barrier, as well as steroidogenesis and proinflammatory cytokines in a model of varicocele rat testes. Materials and Methods: Male rats with experimentally induced varicocele were sacrificed 4 weeks after operation. Testicular histology and blood testosterone concentrations were examined. The expression of tight junctions, steroidogenic enzymes, apoptosis and immune cell markers, and proinflammatory cytokines in the testes were evaluated by reverse transcriptase-polymerase chain reaction, Western blot and immunohistochemistry. Results: Weight and Johnsen scores of varicocele testes were lower than those of normal testes. mRNA expression of Bad and Bax increased whereas Bcl-xl and Bcl2 mRNA decreased in varicocele testes compared to controls. Although blood testosterone did not change, Leydig cell 3bHsd immunoreactivity, testicular 3 beta Hsd6 and 17 beta Hsd3 mRNA were significantly decreased in varicocele testes. Cldn11 mRNA and protein levels in varicocele testes were higher than in normal testes together with altered expression of Ocln, Zo1 and N-cadherin mRNA. Cldn11 immunoreactivity appeared as wavy strands at the periphery of the seminiferous epithelium in normal testes but was frequently found in the Sertoli cell cytoplasm in varicocele testes. In varicocele testes Tnf alpha, Il1 alpha, Il6, Cd45, Cd3g and Cd3d mRNA was increased. Conclusions: An increase in proinflammatory cytokines might be responsible for deregulation of Cldn11 in the Sertoli cells in varicocele testes, leading to alterations in the permeability of the blood-testis barrier and immunological barriers to normal spermatogenesis.</P>

흰쥐에서 일측 고환염전과 염전복구 후의 환측 및 대측 고환과 부고환의 조직학적 변화

백민기,백성현,오승준,정성진,홍성규,박인애,전윤경,최황 대한비뇨의학회 2008 Investigative and Clinical Urology Vol.49 No.10

Purpose: This investigation was undertaken to determine the damage to the testes and epididymides following torsion and detorsion of the testes. Materials and Methods: The right testes of 8-week-old male rats(n=30) were subjected to torsion for 10 min. At 0, 1, 4, 8, and 24 hours, and 1 week after the repair of a torsion, the ipsilateral and contralateral testes and epididymides were harvested. The mean number of spermatids per tubule, the mean seminiferous tubular diameter(MSTD), and the germinal epithelial cell thickness(GECT) were used to evaluate changes to the testes. The histologic changes to the epididymal ductal epithelium were also evaluated. Results: The mean number of spermatids per tubule, GECT, and MSTD were significantly decreased in the 24-hour ipsilateral detorsion group, but minimal changes to ipsilateral testes were observed in the 1-week detorsion group. There was no evidence of histologic changes to the testes in any of the contralateral detorsion groups. The interstitial fibroblast proliferation and hemorrhage of the ipsilateral epididymis were found in the 4-hour detorsion group and increased in the 8-hour detorsion group. Interstitial fibroblast proliferation was prominent in the ipsilateral epididymis of the 24-hour detorsion group, but was only occasionally observed in the contralateral epididymides. Shortening of the tubular epithelial cell height and tubule dilatation were observed in the ipsilateral and contralateral epididymis 1 week after detorsion. Conclusions: Torsion/detorsion damage was found earlier and at a higher intensity in the epididymides than in the testes. This finding may be due to the protection afforded by the blood-testis barrier. Purpose: This investigation was undertaken to determine the damage to the testes and epididymides following torsion and detorsion of the testes. Materials and Methods: The right testes of 8-week-old male rats(n=30) were subjected to torsion for 10 min. At 0, 1, 4, 8, and 24 hours, and 1 week after the repair of a torsion, the ipsilateral and contralateral testes and epididymides were harvested. The mean number of spermatids per tubule, the mean seminiferous tubular diameter(MSTD), and the germinal epithelial cell thickness(GECT) were used to evaluate changes to the testes. The histologic changes to the epididymal ductal epithelium were also evaluated. Results: The mean number of spermatids per tubule, GECT, and MSTD were significantly decreased in the 24-hour ipsilateral detorsion group, but minimal changes to ipsilateral testes were observed in the 1-week detorsion group. There was no evidence of histologic changes to the testes in any of the contralateral detorsion groups. The interstitial fibroblast proliferation and hemorrhage of the ipsilateral epididymis were found in the 4-hour detorsion group and increased in the 8-hour detorsion group. Interstitial fibroblast proliferation was prominent in the ipsilateral epididymis of the 24-hour detorsion group, but was only occasionally observed in the contralateral epididymides. Shortening of the tubular epithelial cell height and tubule dilatation were observed in the ipsilateral and contralateral epididymis 1 week after detorsion. Conclusions: Torsion/detorsion damage was found earlier and at a higher intensity in the epididymides than in the testes. This finding may be due to the protection afforded by the blood-testis barrier.

Histological Analysis and Investigation of Germ Cell Markers in Neonatal and Pubertal Pig Testis

Kyung Hoon Lee,Hee Chan Kim,Won Young Lee,Woo Tae Ha,Ran Lee,Hyuk Song 한국동물번식학회 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s

INTRODUCTION In rodent, molecular markers of spermatogonia, spermatocyte, spermatid and sperm have been identified. It has been reported that DBA, PGP 9.5 and NanpG can be the markers for spermatogonia in pig. For further understanding the spermatogenesis of the pig on morphological and molecular level, we report identification of testicular cells in neonatal and pubertal pig testis, and a putative marker for spermatogonia and spermatid in pig testis. MATERIALS AND METHODS Neonatal (3 day) and pubertal testis (150 day) was cut and fixed in Bouin’s solution for immunohistochemistry (IHC). Gonocytes were isolated from neonatal testis for immunocytochemistry (ICC). Based on references (Phillips et al., 2010), thirteen antibodies (VASA, Oct4, NanoG, PGP 9.5, DAZL, SCF, GFR-alpha 1, PLZF, c-kit, integrin-beta1, Thy1, Sohlh1 and CD9) were used for IHC and ICC. Paraffin section was performed for IHC. Gonocytes were attached to the APS-coated slides for ICC. HRP-conjugated and florescent-labeled secondary antibody was used for IHC and ICC, respectively. RESULTS In histological analysis, spermatogonia and Sertoli cells, which are enclosed by seminiferous tubule and connective tissue, were observed in neonatal testis. However, complete spermiogenesis, including spermatocyte, spermatid and spermatozoa, was not observed in neonatal testis. Spermatocyte, spermatid and elongated spermatid were observed in pubertal testis but matured spermatozoa were not observed. As a result, two antibodies (PGP 9.5 and GFR-alpha1) of thirteen antibodies were available for IHC and ICC. As reported in other studies, PGP 9.5 protein was detected in spermatogonia of ne-onatal in IHC. In addition, it was observed in spermatogonia of pubertal testis. GFR- alpha1 protein was detected in spermatogonia of neonatal testis and spermatid of pubertal testis. In ICC, PGP 9.5 protein was detected in gonocytes as reported in other studies. GFR-alpha1 was also observed in gonocyte isolated from pig testis. In this study, we found that 1) only spermatogonia exist in neonatal pig testis (3 day), 2) GFR-alpha1 is a new marker for spermatogenesis in pig testis.

퇴축고환의 조직병리학적 특징

이택,한상원,이민종,김장환,최승강,조남훈,한지영 大韓泌尿器科學會 1999 Korean Journal of Urology Vol.40 No.5

Diagnostic Laparoscopy for the Management of Impalpable Testes

박지현,박용현,박관진,최황 대한비뇨의학회 2011 Investigative and Clinical Urology Vol.52 No.5

Purpose: Controversy exists regarding the best approach to impalpable testes. We determined the usefulness of diagnostic laparoscopy for the management of impalpable testes. Materials and Methods: Between 2000 and 2008, 86 patients with a mean age of 34 months underwent diagnostic laparoscopy. An inguinal canal exploration was performed in all cases, except in patients in whom the internal spermatic vessels terminated intraperitoneally with a blind end. Results: The undescended testis was right-sided in 24 patients (27.9%), left-sided in 47 patients (54.7%), and bilateral in 15 patients (17.4%). Three patients (3.5%) had bilateral impalpable testes. The vas and vessels traversed the internal ring in 51 of 89 impalpable testes (57.3%); 20 (22.5%) were localized intraperitoneally, and 18 (20.2%) were diagnosed as vanishing testes. Open orchiopexies were performed on 24 testes (27.0%) and orchiectomies were performed on 43 nubbin testes (48.3%). After a mean follow-up period of 30 months, 12 of the 14 testes (85.7%) were viable following open conventional orchiopexy, compared with 6 of the 10 testes (60%) following a 1-stage Fowler-Stephens orchiopexy. Conclusions: Diagnostic laparoscopy is a very helpful and minimally invasive technique in the diagnosis of impalpable testes, especially when preoperative ultrasonography is not sufficiently informative. Purpose: Controversy exists regarding the best approach to impalpable testes. We determined the usefulness of diagnostic laparoscopy for the management of impalpable testes. Materials and Methods: Between 2000 and 2008, 86 patients with a mean age of 34 months underwent diagnostic laparoscopy. An inguinal canal exploration was performed in all cases, except in patients in whom the internal spermatic vessels terminated intraperitoneally with a blind end. Results: The undescended testis was right-sided in 24 patients (27.9%), left-sided in 47 patients (54.7%), and bilateral in 15 patients (17.4%). Three patients (3.5%) had bilateral impalpable testes. The vas and vessels traversed the internal ring in 51 of 89 impalpable testes (57.3%); 20 (22.5%) were localized intraperitoneally, and 18 (20.2%) were diagnosed as vanishing testes. Open orchiopexies were performed on 24 testes (27.0%) and orchiectomies were performed on 43 nubbin testes (48.3%). After a mean follow-up period of 30 months, 12 of the 14 testes (85.7%) were viable following open conventional orchiopexy, compared with 6 of the 10 testes (60%) following a 1-stage Fowler-Stephens orchiopexy. Conclusions: Diagnostic laparoscopy is a very helpful and minimally invasive technique in the diagnosis of impalpable testes, especially when preoperative ultrasonography is not sufficiently informative.

비촉지성 정류고환 97례의 수술경험

이동형,김광세 大韓泌尿器科學會 2001 Korean Journal of Urology Vol.42 No.11

비촉지성 정류고환에서 진단적 복강경검사의 유용성

이영국,박재신,이은석,Lee, Young-Guk,Park, Jae-Shin,Lee, Eun-Suk 대한소아외과학회 2007 소아외과 Vol.13 No.1

There have been no definitive preoperative diagnostic imaging studies for impalpable testes. We observed the effectiveness of laparoscopy for detecting impalpable testes not identified with ultrasonography (USG) or careful physical examination under general anesthesia. We retrospectively reviewed 117 patients (118 testes) who were operated upon for undescended testes from January 1998 to December 2004. The testes of these patients were palpable in 97(82 %) and impalpable in 21 (18 %). We analyzed the preoperative diagnostic method, site of the testes, operative method and operative findings of the 21 impalpable testes. Preoperative USG and physical examination under general anesthesia were performed on 20 patients, and 12 patients' testes could be localized. Eight patients whose testes could not be localized with USG and physical examination underwent laparoscopy. Seven of the 8 patients had testes in inguinal canal and 4 of these were atrophied and underwent orchiectomy because of atrophy (2) and vanishing (2). Only 1 patient had bilateral intraabdominal testes and one of the testes was atrophied. Laparoscopy was a useful method for detecting impalpable testes, but the clinical application might be limited because the location of atrophic or vanishing testes was mainly inferior to internal inguinal ring.

Profiling of testis-specific or testis-predominant genes expressed in mouse male germ cell lines GC-1 and GC-2

Choi Seungho,Hong Seong Hyeon,Han Gwidong,Cho Chunghee 한국유전학회 2024 Genes & Genomics Vol.46 No.3

Background Spermatogenesis is a tightly organized process that utilizes an intrinsic genetic program composed of germ cell-specific genes. Although mouse germ cell-related cell lines are available, few germ cell-specific genes have been comprehensively identified in such cell lines. Objective We aimed to profile gene expression in the male mouse germ cell-related cell lines, GC-1 and GC-2, characterize their transcriptomic nature, and identify potential testis- or germ cell-specific or -predominant genes expressed in these cell lines. Methods We performed profiling analysis of genes transcribed in the mouse germ cell-related cell lines, GC-1 and GC-2, using our previous microarray data together with public transcriptome information. We analyzed the expression of a number of the cell line genes predicted to be preferentially expressed in testis by RT-PCR. Results We found that most testis-specific or -predominant mRNAs are not expressed in GC-1 and GC-2 cells, implying that these cell lines have lost their testis- or germ cell-specific genetic characteristics. RT-PCR analysis of genes predicted to be expressed in the cell lines with preferential testicular expression showed the testis-specific or -predominant expression of nine genes and verified four of them as being expressed in the germ cell lines. Among them, only cyclin-dependent kinase inhibitor 3 genes (Cdkn3) showed testis and germ cell specificity. Conclusion Our study provides extensive transcriptomic information to shed light on the limited testicular characteristics of the mouse male germ cell-derived cell lines, GC-1 and GC-2, and offers a list of germ cell line genes with testicular preference. Background Spermatogenesis is a tightly organized process that utilizes an intrinsic genetic program composed of germ cell-specific genes. Although mouse germ cell-related cell lines are available, few germ cell-specific genes have been comprehensively identified in such cell lines. Objective We aimed to profile gene expression in the male mouse germ cell-related cell lines, GC-1 and GC-2, characterize their transcriptomic nature, and identify potential testis- or germ cell-specific or -predominant genes expressed in these cell lines. Methods We performed profiling analysis of genes transcribed in the mouse germ cell-related cell lines, GC-1 and GC-2, using our previous microarray data together with public transcriptome information. We analyzed the expression of a number of the cell line genes predicted to be preferentially expressed in testis by RT-PCR. Results We found that most testis-specific or -predominant mRNAs are not expressed in GC-1 and GC-2 cells, implying that these cell lines have lost their testis- or germ cell-specific genetic characteristics. RT-PCR analysis of genes predicted to be expressed in the cell lines with preferential testicular expression showed the testis-specific or -predominant expression of nine genes and verified four of them as being expressed in the germ cell lines. Among them, only cyclin-dependent kinase inhibitor 3 genes (Cdkn3) showed testis and germ cell specificity. Conclusion Our study provides extensive transcriptomic information to shed light on the limited testicular characteristics of the mouse male germ cell-derived cell lines, GC-1 and GC-2, and offers a list of germ cell line genes with testicular preference.

Long-Term Outcomes of Retractile Testis

배재준,김범수,정성광 대한비뇨의학회 2012 Investigative and Clinical Urology Vol.53 No.9

Purpose: Retractile testis is considered to be a variant of normal testis in prepubertal boys. There is no agreed-upon management of retractile testis. The aim of this study was to provide data on the long-term outcomes of patients with retractile testis. Materials and Methods: This study retrospectively reviewed the medical record of 43 boys who were referred for suspected undescended or retractile testis and were finally diagnosed with retractile testis between January 2001 and December 2008. All boys were biannually examined by a pediatric urologist to evaluate the presence of retractile, descended, or undescended testis and testicular volume. Results: Of 43 boys, there were 22 boys with unilateral retractile testis (51.1%) and 21 boys with bilateral retractile testis (48.9%). Their mean age was 3.0±2.7 years and the follow-up duration was 4.4±1.7 years. Of 64 retractile testes, 29 (45.3%) succeeded in descending, 26 (40.6%) remained retractile, and 9 (14.1%) became undescended testis or of a decreased size requiring orchiopexy. The mean initial diagnostic age of the patients who underwent orchiopexy was 1.3±0.9 years; meanwhile, the mean initial diagnostic age of those who went on to have normal testis was 4.3±3.3 years (p=0.009). The mean follow-up duration was 3.6±1.5 years in the orchiopexy group, 4.0±1.4 years in the descended testis group, and 5.1±1.8 years in group with remaining retractile testis. Conclusions: Retractile testis has a risk of requiring orchiopexy. The risk is higher in the population diagnosed at a younger age. Boys with retractile testis should be observed periodically until the testis is descended in the normal position. Purpose: Retractile testis is considered to be a variant of normal testis in prepubertal boys. There is no agreed-upon management of retractile testis. The aim of this study was to provide data on the long-term outcomes of patients with retractile testis. Materials and Methods: This study retrospectively reviewed the medical record of 43 boys who were referred for suspected undescended or retractile testis and were finally diagnosed with retractile testis between January 2001 and December 2008. All boys were biannually examined by a pediatric urologist to evaluate the presence of retractile, descended, or undescended testis and testicular volume. Results: Of 43 boys, there were 22 boys with unilateral retractile testis (51.1%) and 21 boys with bilateral retractile testis (48.9%). Their mean age was 3.0±2.7 years and the follow-up duration was 4.4±1.7 years. Of 64 retractile testes, 29 (45.3%) succeeded in descending, 26 (40.6%) remained retractile, and 9 (14.1%) became undescended testis or of a decreased size requiring orchiopexy. The mean initial diagnostic age of the patients who underwent orchiopexy was 1.3±0.9 years; meanwhile, the mean initial diagnostic age of those who went on to have normal testis was 4.3±3.3 years (p=0.009). The mean follow-up duration was 3.6±1.5 years in the orchiopexy group, 4.0±1.4 years in the descended testis group, and 5.1±1.8 years in group with remaining retractile testis. Conclusions: Retractile testis has a risk of requiring orchiopexy. The risk is higher in the population diagnosed at a younger age. Boys with retractile testis should be observed periodically until the testis is descended in the normal position.

사춘기와 성년기 흰쥐에서 고환염전에 의한 대측고환 손상 차이에 대한 연구

정성진,최우석,하승범,정민수,정재승,백민기,홍성규,최황 대한비뇨의학회 2009 Investigative and Clinical Urology Vol.50 No.7

Purpose: Our study aimed to determine whether the severity of damage to the contralateral testis by ipsilateral testicular torsion/detorsion in pubertal rats, which have an incomplete blood-testis barrier, is different from that in adult rats. Materials and Methods: We divided pubertal (6 weeks, n=17) and adult (10 weeks, n=17) Sprague-Dawley (SD) rats into group S (sham; n=5), group O (orchiectomy; n=6), and group D (detorsion; n=6). After 4 hours' torsion of the ipsilateral testis, we applied orchiectomy (group O) and detorsion (group D) depending on the group and compared the histopathologic changes and germ cell apoptosis of the contralateral testis at the age of 13 weeks. Results: In each age group, increased interstitial area, edema, and germ cell sloughing were observed in group D. The mean seminiferous tubule diameter decreased more in group D than in group S or O in each age group (p<0.05). The mean germ cell layer thickness and number of spermatids per tubule decreased more in group D than in group S or O in each age group; additionally, in group D, values decreased more in pubertal rats than in adult ones (p<0.05, respectively). The mean numbers of terminal deoxyuridine nick-end labeling (TUNEL)-positive cells were less than 1.0 in groups S and O, which was smaller than in group D (p<0.05); additionally, in group D, this value tended to be higher in pubertal rats than in adult ones (p=0.057). Conclusions: SD rats with a detorsioned testis had more severe damage to the contralateral testis than did those undergoing orchiectomy of the torsioned testis. Also, when comparing the severity of damage to the contralateral testis after ipsilateral torsion/detorsion between pubertal and adult rats, rats at a pubertal age, when most testicular torsions occur in clinical situations, had more severe damage than did those at an adult age. Purpose: Our study aimed to determine whether the severity of damage to the contralateral testis by ipsilateral testicular torsion/detorsion in pubertal rats, which have an incomplete blood-testis barrier, is different from that in adult rats. Materials and Methods: We divided pubertal (6 weeks, n=17) and adult (10 weeks, n=17) Sprague-Dawley (SD) rats into group S (sham; n=5), group O (orchiectomy; n=6), and group D (detorsion; n=6). After 4 hours' torsion of the ipsilateral testis, we applied orchiectomy (group O) and detorsion (group D) depending on the group and compared the histopathologic changes and germ cell apoptosis of the contralateral testis at the age of 13 weeks. Results: In each age group, increased interstitial area, edema, and germ cell sloughing were observed in group D. The mean seminiferous tubule diameter decreased more in group D than in group S or O in each age group (p<0.05). The mean germ cell layer thickness and number of spermatids per tubule decreased more in group D than in group S or O in each age group; additionally, in group D, values decreased more in pubertal rats than in adult ones (p<0.05, respectively). The mean numbers of terminal deoxyuridine nick-end labeling (TUNEL)-positive cells were less than 1.0 in groups S and O, which was smaller than in group D (p<0.05); additionally, in group D, this value tended to be higher in pubertal rats than in adult ones (p=0.057). Conclusions: SD rats with a detorsioned testis had more severe damage to the contralateral testis than did those undergoing orchiectomy of the torsioned testis. Also, when comparing the severity of damage to the contralateral testis after ipsilateral torsion/detorsion between pubertal and adult rats, rats at a pubertal age, when most testicular torsions occur in clinical situations, had more severe damage than did those at an adult age.