Environmental oestrogens disrupt testicular descent and damage male reproductive health: Mechanistic insight.

Environmental oestrogens (EEs) as environmental pollutants have been paid much attention due to their impact on congenital malformation of male genitourinary system. Exposure to EEs for prolonged time could hinder testicular descent and cause testicular dysgenesis syndrome. Therefore, it is urgent to understand the mechanisms by which EEs exposure disrupt testicular descent. In this review, we summarize recent advances in our understanding of the process of testicular descent, which is regulated by intricate cellular and molecular networks. Increasing numbers of the components of these networks such as CSL and INSL3 are being identified, highlighting that testicular descent is a highly orchestrated process that is essential to human reproduction and survival. The exposure to EEs would lead to the imbalanced regulation of the networks and cause testicular dysgenesis syndrome such as cryptorchidism, hypospadias, hypogonadism, poor semen quality and testicular cancer. Fortunately, the identification of the components of these networks provides us the opportunity to prevent and treat EEs induced male reproductive dysfunction. The pathways that play an important role in the regulation of testicular descent are promising targets for the treatment of testicular dysgenesis syndrome.

Intestinal obstruction caused by mesodiverticular band in children: a cohort study.

OBJECTIVE: To explore clinical characteristics, pathogenesis, diagnosis and treatment of intestinal obstruction due to mesodiverticular band (MDB) in children in a single center in China. METHODS: The clinical data of 20 children with acute intestinal obstruction due to MDB between 1998 and 2020 were retrospectively analyzed. RESULTS: The male-to-female ratio was 14:6 in 20 cases. Except one case of 7-month pregnant stillbirth, the cases were aged from 7 days to 14 years, at the median age of 4.31 years. The common symptoms were vomiting, abdominal pain and/or abdominal distension. About 40% (8/20) of patients had both MDB and Meckel's Diverticulum (MD), while 60% (12/20) of patients had MDB only. Only one case died because of total colonic aganglionosis, while other children recovered after surgery treatment. MDB led to the strangulation of necrotic bowel in six cases, intestinal perforation in one case, and intestinal rupture in one case. Pathologic examination showed thick-walled arteries and or thick venous vascular structures in the cord. All cases had no complications during 1-year follow-up. CONCLUSION: MDB results from the remnant of vitelline vessel, and often causes acute intestinal obstruction without special clinical symptoms. Unexplained abdominal pain and distension without surgery history should be paid attention, especially for strangulated intestinal obstruction. Timely surgical exploration is beneficial to avoid intestinal necrosis or even sudden death, and the pathological examination is important for the diagnosis.

The RXFP2-PLC/PKC signaling pathway mediates INSL3-induced regulation of the proliferation, migration and apoptosis of mouse gubernacular cells.

BACKGROUND: Testicular hypoplasia can affect the sexual and reproductive ability in adulthood, and even increase the risk of cancer. Abnormal development of the gubernaculum is one of the important factors of testicular hypoplasia. Therefore, a study of the structure and function of the gubernaculum is an important but neglected new breakthrough point for investigating the normal/abnormal development of the testis. Previous findings showed that Insulin like factor 3 (INSL3) is a key factor regulating the growth of gubernaculum, however, the mechanism by which INSL3 acts on the gubernaculum remains unknown. Therefore, we probed the mechanism associated with INSL3-induced the proliferation, migration, and apoptosis of gubernacular cells in mice. METHODS: A culture cell model of neonatal mice gubernaculum is established by INSL3 intervention. We blocked PLC/PKC signaling pathway with U73122 pretreat to investigate the role of the PLC/PKC signaling pathway. The changes of cell proliferation, migration, and apoptosis were detected by molecular biological methods. In addition, the levels of PCNA and F-action were detected by immunofluorescence and western blotting. RESULTS: We found that INSL3 can promote the proliferation and migration of gubernacular cells and inhibit their apoptosis, meanwhile, INSL3 significantly up-regulated PLC/PKC protein phosphorylation. However, treatment with the PLC/PKC signaling pathway inhibitor U73122 significantly inhibited these effects of INSL3. Besides, we found that INSL3 could up-regulate the protein expression level of PCNA and F-actin, while the PCNA and F-actin expression was significantly weakened after U73122 pretreatment. CONCLUSIONS: This research revealed that INSL3 binding to RXFP2 may up-regulate the expression levels of PCNA and F-actin by activating the PLC/PKC signaling pathway to promote the proliferation and migration of gubernacular cells. It suggests that the RXFP2-PLC/PKC axis may serve as a novel molecular mechanism by which INSL3 regulates growth of the gubernaculum.

Risk factors and predictive model for abdominal wound dehiscence in neonates: a retrospective cohort study.

BACKGROUND: Abdominal wound dehiscence (AWD) is a major complication of abdominal surgery, and neonates are a group with a high risk of AWD, which has serious consequences or can even result in death. The purpose of this study is to explore the risk factors for neonatal AWD and construct a predictive model. METHODS: The clinical data of 453 cases that underwent neonatal laparotomy from June 2009 to June 2020 were retrospectively analyzed, among which 27 cases of AWD were identified. Nine factors, including gender, age at admission, weight at admission, preterm delivery, level of preoperative anaemia, hypoalbuminemia, operation time, incision length, and incision type, were analyzed to explore their correlation with neonatal AWD. RESULTS: The incidence of neonatal AWD was 6.0% (27/453), among which partial wound dehiscence accounted for 4.9% (22/453) and complete wound dehiscence accounted for 1.1% (5/453). Hypoproteinemia and incision type were the independent risk factors for neonatal AWD, and weight at admission was a protective factor for AWD in the multivariate models. All these factors were incorporated to construct a nomogram, and a calibration curve was plotted. The result indicated that the actual risk was close to the predicted risk when the predicted risk rate was greater than about 35%. CONCLUSIONS: Neonatal AWD is closely related to hypoproteinemia and incision contamination. Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.Key messagesNeonatal abdominal wound dehiscence (AWD) has a serious consequence and the incidence of neonatal AWD was about 6.0% and the complete AWD morbidity is 1.1%.Hypoproteinemia and incision type were the independent risk factors for neonatal AWD.Our predictive model showed the potential to provide an individualized risk estimate of AWD for neonatal patients undergoing abdominal surgery.

RXFP2 as novel potential biomarker for abnormal differentiation induced by diethylstilbestrol in the gubernaculum of fetal mice.

Environmental estrogens (EEs) have been correlated with abnormalities in the male urogenital system. However, the mechanism underlying the effect of these molecules remains unclear. In vitro and in vivo experiments were performed to examine the expression level and mechanism of relaxin family peptide receptor 2 (RXFP2) in the gubernaculum of fetal mice following diethylstilbestrol (DES) treatment. The in vivo results demonstrate that DES treatment increased the stillbirth rate gradually, decreased the gubernacular cone volume significantly, and disrupted the tissue structure, leading to incomplete testicular descent. In vitro experiments reveal that DES administration resulted in abnormal cellular morphology and structural disorder of gubernacular cells, which lost their original morphology in a dose-dependent manner. Moreover, DES-induced F-actin rearrangement and stress fiber formation in cultured cells. Protein quantitative analysis showed that the RXFP2 level in each experimental group was significantly lower than that of the normal group. In conclusion, DES affects the morphology and alters the gubernaculum structure, as well as the expression of RXFP2 protein. These data demonstrate that DES is toxic to gubernaculum in fetal mice, and that RXFP2 is associated with the abnormal gubernaculum morphology induced by DES. Taken together, these data suggest that RXFP2 may be a novel potential biomarker for abnormal differentiation of the gubernaculum.

Diagnosis and Treatment of Hypospadias With Megameatus Intact Prepuce.

Purpose: To evaluate the diagnosis and treatment methods of hypospadias with megameatus intact prepuce (MIP). Materials and Methods: A retrospective analysis was performed in 27 MIP children, 13 of whom underwent tubularized incised plate urethroplasty (TIP procedure), 7 underwent the Duplay procedure, 5 underwent the Mathieu procedure, 1 underwent meatal advancement and glanuloplasty (MAGPI procedure), and 1 underwent the glans approximation procedure (GAP). The patients were followed for 6-36 months to evaluate the surgical outcomes by the Pediatric Penile Perception Score (PPPS). Results: A total of 27 patients with a mean age of 8.12 ± 3.0 years were enrolled in this study, and 25 cases (25/27, 92.6%) were accidentally discovered during the first visit for phimosis. The patients had a formed urethra of 0.5 to 1.5 cm. Complications occurred in 4 of the 27 patients (14.81%): 2 patients with urethral fistula and 2 patients with meatal stenosis. One patient had a case of self-healed urethral fistula, and the remaining 3 patients underwent reoperation. The post-operative effect was satisfactory in all patients, and the urinary flow and stream during urination were normal. The overall average PPPS score of non-operative surgeons and parents was satisfactory. There were no significant differences in meatus appearance, glans appearance, skin appearance, and general appearance PPPS score among the Mathieu, TIP, and Duplay surgical procedures. Conclusions: MIP clinical manifestations are concealed and usually noted when circumcision is attempted. The suitable procedure for each patient should be tailored according to the anatomic features, and several techniques can be used with good functional and cosmetic results.

Megameatus intact prepuce treated with urethral plate-preserving surgery: a retrospective study of an unusual hypospadias variant.

BACKGROUND: Megameatus intact prepuce (MIP) is a unique variant of hypospadias and is a clinically rare condition. Due to the anatomical characteristics of the MIP hypospadias variant presenting a unique challenge to surgeons, no single urethroplasty method provides a universal solution for all patients. The purpose of this study was to evaluate the outcomes of hypospadias after MIP repair by urethral plate-preserving urethroplasty. METHODS: A retrospective study was performed on 25 coronal or distal MIP patients, with a median age of 8, with most deficiencies being discovered during their first hospital visit for phimosis. Correction with urethroplasty was performed for all patients; 5 underwent the Mathieu procedure, 13 underwent the tubularized incised plate (TIP) procedure, and 7 underwent the Duplay procedure. The 25 patients were followed up for 6 to 36 months to evaluate the surgical outcomes. RESULTS: There were no significant differences in intraoperative bleeding, hospital stays, postoperative analgesia rate, and cure rate among the three surgical procedures. The operative time for the Mathieu procedure was longer than that for the TIP and Duplay procedures, which did not differ. Complications occurred in 4 of the 25 patients (16.0%), and the overall complication-free survival rate at 1 year after surgery was 80.5%. The age at the time of surgery, urethral plate width, urethroplasty length, surgical procedures, or meatal location (coronal or distal penis) were not independently predictive of complications. CONCLUSIONS: The clinical manifestations of MIP are often concealed and then accidentally discovered during hospital visits for phimosis; thus, the actual incidence of MIP might be higher. The urethral plate should be preserved during MIP-correcting treatment, especially for coronal or distal MIP. The same satisfactory outcomes can be obtained with Mathieu, TIP, or Duplay urethroplasty.

Children with Cryptorchidism Complicated by Testicular Torsion: A Case Series.

OBJECTIVE: To investigate the clinical features, diagnosis, treatment and prognosis of children with cryptorchidism complicated by testicular torsion. METHODS: The clinical data of 6 children with cryptorchidism complicated by testicular torsion admitted to our hospital from December 2000 to December 2016 were analyzed retrospectively. RESULTS: All 6 children were diagnosed with cryptorchidism by surgery, their age was from 12 days up to 11 years, and the average time between onset of symptoms and diagnosis was 20.5 h. Torsion testis was located in the groin area and the rate of left to right was about 2:1. Twist was 600° on average. All children were admitted because of the inconsolable cry, abdominal pain, and the swelling of the groin. Three patients underwent orchidectomy, while the other 3 patients underwent detorsion and cryptorchidopexy. Color Doppler ultrasound examination showed normal testes at 6 months after operation. Only 1 case was diagnosed with cryptorchidism after birth. CONCLUSIONS: Cryptorchidism is an emergency in pediatric urology and often leads to a low testicular salvage rate, especially in infants, due to lack of knowledge, delayed diagnosis, and late treatment. Neonatal genital examination is important for the early diagnosis and management of cryptorchidism.

Diethylstilbestrol regulates mouse gubernaculum testis cell proliferation via PLC-Ca2+ -CREB pathway.

Recent evidence suggested a positive correlation between environmental estrogens (EEs) and high incidence of abnormalities in male urogenital system, but the mechanism remains unclear. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the morphology and proliferation of gubernaculum testis cells, but the underlying mechanism is unclear. In this study, mouse gubernaculum testis cells were pretreated with phospholipase C (PLC) inhibitor U-73122 and then treated with DES. The results demonstrated that U-73122 impaired DES-evoked intracellular Ca2+ mobilization in gubernaculum testis cells and inhibited DES-induced proliferation of gubernaculum testis cells. Mechanistically, we found that U-73122 inhibited DES-induced activation of cAMP-response element binding protein (CREB) in gubernaculum testis cells. In conclusion, these data suggest that the effects of DES on mouse gubernaculum testis cells are mediated by PLC-Ca2+ -CREB pathway. SIGNIFICANCE OF THE STUDY: Environmental estrogens remain a serious threat to male reproductive health, and it is important to understand the mechanism by which EEs affect the male productive system. Here we explore potential mechanisms how the proliferation and contractility of gubernaculum testis cells are regulated by diethylstilbestrol. Our findings provide the first evidence that PLC-Ca2+ -CREB signalling pathway mediates the nongenomic effects of diethylstilbestrol on gubernaculum testis cells. These findings provide new insight into the role of diethylstilbestrol in the aetiology of male reproductive dysfunction and will help develop better approaches for the prevention and therapy of male reproductive malformation.

[Impacts of DES on the expressions of related genes in the gubernaculums testis of newborn mice].

OBJECTIVE: To investigate the influence of diethylstilbestrol (DES) on the mRNA expressions of the androgen receptor (AR), estrogen receptor α (ERα), proliferating cell nuclear antigen (PCNA), and actin alpha 1 (ACTα1) in the gubernaculums testis of newborn mice and explore their action mechanisms. METHODS: A total of 140 male Kunming mice were randomly divided into a blank control, a dimethyl sulfoxide (DMSO) control, and 5 experimental groups to be treated subcutaneously with normal saline, DMSO, and DES at 0.02, 0.1, 0.5, 10 and 50 µg per kg of the body weight per day, respectively, at gestation days 9－17. On the first day after birth, the animals were sacrificed and the gubernaculums testis collected for detection of the mRNA expressions of AR, ERα, PCNA and ACTα1 by RT-PCR. RESULTS: Compared with the DMSO control, the experimental groups, particularly the DES 10 and 50 µg groups, showed significant increases in the mRNA expression of ERα (RE2 = 0.825, P <0.05), but remarkable decreases in those of AR, PCNA and ACTα1 (RA2 = 0.713, RP2 = 0.946, RT2 = 0.960, P <0.01), all in a dose-dependent manner. CONCLUSIONS: The AR, ERα, PCNA, and ACTα1 mRNA are expressed in the gubernaculum testis of normal newborn mice, and their expression levels may be influenced by intervention with different concentrations of DES during the gestation. Exogenous estrogens may affect the proliferation and contraction of gubernaculum testis cells and consequently the normal development of the testis or even the whole male reproductive system by influencing the metabolism of ER and/or AR.

[Roles of G protein-coupled estrogen receptor in the male reproductive system].

The G protein-coupled estrogen receptor (GPER), also known as G protein-coupled receptor 30 (GPR30), was identified in the recent years as a functional membrane receptor different from the classical nuclear estrogen receptors. This receptor is widely expressed in the cortex, cerebellum, hippocampus, heart, lung, liver, skeletal muscle, and the urogenital system. It is responsible for the mediation of nongenomic effects associated with estrogen and its derivatives, participating in the physiological activities of the body. The present study reviews the molecular structure, subcellular localization, signaling pathways, distribution, and function of GPER in the male reproductive system.

Diethylstilbestrol Regulates the Expression of LGR8 in Mouse Gubernaculum Testis Cells.

BACKGROUND Hormonal effects on the gubernaculum can affect testicular descent. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the outgrowth of gubernaculums, leading to testis maldescent. However, the underlying mechanisms remain elusive. MATERIAL AND METHODS The gubernaculum were removed from 3-day-old mice and cultured. The subcultured cells were randomly divided into a normal control group and experimental groups. The DES groups were administered 10 µg/ml, 1 µg/ml, 0.1 µg/ml, 0.01 µg/ml of diethylstilbestrol dissolved in dimethyl sulfoxide (DMSO) respectively. The cell morphology was observed under an inverted microscope, and leucine-rich repeat-containing G protein-coupled receptor 8 (LGR8) was localized by immunofluorescence. The expressions of LGR8 gene and protein in gubernaculum cells were quantified by RT-PCR and Flow Cytometer respectively. RESULTS DES treatment converted cells from a normal fibroblast-like morphology into a more refractile, spindle-shaped morphology or irregular elliptical shapes along with cytoplasmic shrinkage. LGR8 was expressed in the cytoplasmic membrane, DES dose-dependently downregulated LGR8 expression at low doses (≤1.0 µg/ml), but upregulated LGR8 at high doses (10 µg/ml) at both the mRNA and protein levels. CONCLUSIONS These results suggest that DES causes testicular maldescent by altering the LGR8 pathway in mouse gubernaculum testis cells.

Altered Levels of Zinc and N-methyl-D-aspartic Acid Receptor Underlying Multiple Organ Dysfunctions After Severe Trauma.

BACKGROUND: Severe trauma can cause secondary multiple organ dysfunction syndrome (MODS) and death. Oxidative stress and/or excitatory neurotoxicity are considered as the final common pathway in nerve cell injuries. Zinc is the cofactor of the redox enzyme, and the effect of the excitatory neurotoxicity is related to N-methyl-D-aspartic acid receptor (NMDAR). MATERIAL AND METHODS: We investigated the levels of zinc and brainstem NMDAR in a rabbit model of severe trauma. Zinc and serum biochemical profiles were determined. Immunohistochemistry was used to detect brainstem N-methyl-D-aspartic acid receptor 1 (NR1), N-methyl-D-aspartic acid receptor 2A (NR2A), and N-methyl-D-aspartic acid receptor 2B (NR2B) expression. RESULTS: Brain and brainstem Zn levels increased at 12 h, but serum Zn decreased dramatically after the trauma. NR1 in the brainstem dorsal regions increased at 6 h after injury and then decreased. NR2A in the dorsal regions decreased to a plateau at 12 h after trauma. The levels of NR2B were lowest in the death group in the brainstem. Serum zinc was positively correlated with NR2A and 2B and negatively correlated with zinc in the brain. Correlations were also found between the brainstem NR2A and that of the dorsal brainstem, as well as between brainstem NR2A and changes in NR2B. There was a negative correlation between zinc and NR2A. CONCLUSIONS: Severe trauma led to an acute reduction of zinc enhancing oxidative stress and the changes of NMDAR causing the neurotoxicity of the nerve cells. This may be a mechanism for the occurrence of MODS or death after trauma.

Diethylstilbestrol affects the expression of GPER in the gubernaculum testis.

Recent evidence suggested a positive correlation between environmental estrogens (EEs) and high incidence of abnormalities in male urogenital system. EEs are known to cause the abnormalities of testes development and testicular descent. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the morphology and proliferation of gubernacular cells, and its nongenomic effects on gubernaculum testis cells may be mediated by G protein-coupled estrogen receptor (GPER). In this study, we detected the expression of GPER in mouse gubernacular testis and investigated the effects of DES on the expression of GPER in gubernaculum testis cells. RT-PCR analysis revealed that GPER mRNA was expressed in the gubernaculum. GPER protein was detected in the parenchymal cells of the gubernaculum early in development. Furthermore, we demonstrate that GPER inhibitor G15 relieved DES-induced inhibition of GPER expression in gubernaculum testis cell, but ER inhibitor ICI 182780 had the converse effects on DES-induced inhibition of GPER expression in these cells. These data suggest that the effects of DES on mouse gubernaculum testis cells are mediated at least partially by the regulation of GPER expression.

Computer-aided three-dimensional reconstruction of main vessels in hemangiomas.

This study aimed to investigate three-dimensional (3-D) morphological features of the main vessel architecture of human hemangioma. Serial sections of specimens from three cases of children hemangioma were stained with hematoxylin and eosin (HE) to visualize the vessels. Serial images were taken and processed with computer-assisted 3-D reconstruction. Partial 3-D structure reconstruction of vessel morphology in hemangioma revealed strange distribution and branching, which were different from normal vessels of the human skin. The 3-5 microvascular was most common in hemangioma. We observed respective characteristics of three cases: 1 case showed uniform artery vein distribution accompanied by running trend; 1 case showed main artery distribution and less vein distribution, and there were many blood sinus in the shallow surface close to the skin; another case showed vein distribution in the middle of antrum. In conclusion, digital vascular model of 3-D structure of main vessel hemangioma provides a new way for the diagnosis and treatment of hemangioma of children.

Blood zinc, iron, and copper levels in critically ill neonates.

The aim of this study is to explore the prognostic value of blood zinc, iron, and copper levels in critically ill neonates by comparing blood metal levels with the score for neonatal acute physiology (SNAP). Forty-six neonates (26 boys, 20 girls; ages ranging from 10 min to 23 days) who had been admitted to the neonatal intensive care unit of hospital and who were critically ill according to SNAP were included. Another 15 neonates (12 boys, 8 girls; ages ranging from 30 min to 24 days) who were brought to the hospital for a health checkup were included as controls. Clinical data, time in the intensive care unit, prognosis, and SNAP for critically ill neonates were recorded. Blood Cu, Zn, and Fe values were measured by inductively coupled plasma atomic emission spectrophotometry. Ill neonates were divided into extremely critical (SNAP ≥ 10) and critical groups (1 ≤ SNAP < 9). Zn levels were lower in patients than in controls (p <0.05). Cu levels did not differ between patients and controls (p >0.05). Fe levels were not significantly between the critical and control groups (p >0.05). In ill neonates, blood Zn and Fe concentrations in the extremely critical group were lower than in the critical group (p <0.05). Serious illness in neonates may lead to decreased Zn and Fe blood concentrations. Zn and Fe supplements may be beneficial for critically ill children.

An insight into insulin-like factor 3 regulate its receptor RXFP2 in mouse gubernaculum testis cells.

The etiology of testicular dysgenesis syndrome is multifactorial and involves abnormalities in the anatomical structures and endocrine factors. Several studies have shown that the abnormal development of the gubernaculum may affect testicular descent, and the insulin-like factor 3 (INSL3) appears to play an important role in development of the gubernaculum have been proved. INSL3 binds its specific receptor (Relaxin family peptide 2, RXFP2), which was highly expressed in gubernaculum, to produce a crucial effect in the first transabdominal descent stage, but its mechanism still remain unclear. In this study, in order to explore how does INSL3 regulate its receptor RXFP2, we cultured mouse gubernaculum testis cells in vitro, which was treated by INSL3, and examined the expression of RXFP2 in mouse gubernaculum testis cells. The results displayed that INSL3 changed RXFP2 expression, and we found that low dose INSL3 can increase RXFP2 expression, the mechanism of above-mentioned might be related with the hormesis of INSL3.

The composition of surgical teams in the operating room and its impact on surgical team performance in China.

BACKGROUND: Previous studies on surgical team composition have shown that surgical team size had an independent impact on surgical performance in US and Canadian hospitals. We aimed to investigate the impact of team composition on surgical performance in two Chinese hospitals. METHODS: General surgery procedures performed from April 2011 to June 2012 were retrospectively reviewed to record the number of attendees in the operating room (OR) and the procedure time (PT). RESULTS: A total of 1,900 valid procedures, mostly laparoscopic, were performed during the study period. The mean PT was 90.5 min. On average, there were a total of 6 (range = 3-8) team members per procedure: 3 (range = 1-5) surgeons, 2 nurses, and 1 anesthesiologist. Unlike the data reported for the US and Canada, the number of nurses and anesthesiologists remained stable in most cases, whereas the number of surgeons differed by procedure. Multiple-regression analysis revealed that both the complexity of the operation and the team size significantly affected PT. When procedure complexity and patient condition were kept constant, adding one team member in our data analysis predicted an increase of 34.7 min in the PT. CONCLUSION: The surgical team size has a measurable effect on PT. Aside from surgical complexity, the team composition and member stability affected PT in the OR. Optimizing surgical teams and developing a strategy to maintain team stability are of great importance for improving OR efficiency.

G protein-coupled estrogen receptor-protein kinase A-ERK-CREB signaling pathway is involved in the regulation of mouse gubernaculum testis cells by diethylstilbestrol.

The etiology of testicular dysgenesis syndrome is multifactorial and involves environmental factors, such as environmental estrogens. Several studies have shown that hormonal effects on the gubernaculum may affect testicular descent. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the morphology and proliferation of gubernacular cells, but the underlying mechanisms remain elusive. In this study, we aimed to determine whether DES may regulate the function of gubernaculum testis cells by way of nongenomic effects mediated by G protein-coupled estrogen receptor (GPER). We used cultured mouse gubernacular testis cells to demonstrate that GPER is expressed in gubernaculum testis cells. Erk1/2 inhibitor PD98059, PKA inhibitor H89, and Src inhibitor PP2 relieved DES-induced inhibition of gubernaculum testis cell proliferation, but ER inhibitor ICI 182780 had no effects on DES-induced inhibition of gubernaculum testis cell proliferation. In addition, we found that DES induced the activation of CREB downstream of PKA, Src, and ERK1/2 in these cells. These data suggest that the effects of DES on mouse gubernaculum testis cells are mediated at least partially by GPER-protein kinase A-ERK-CREB signaling pathway.

Diethylstilbestrol impairs the morphology and function of mouse gubernaculum testis in culture.

Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen widely used in estrogen therapy. In animal models, exposure to DES disrupts the outgrowth of the gubernacula, leading to testis maldescent. However, it remains unclear whether the effects of DES on gubernaculum are direct or indirect, and the underlying mechanisms are largely obscure. In this study, mouse gubernaculum testis cells were isolated and treated by DES, and cell morphology and function were examined. The results showed that DES changed the morphology and inhibited the proliferation of gubernacular cells. Furthermore, DES increased intracellular [Ca(2+)] and induced F-actin rearrangement and stress fiber formation in gubernaculum testis cells in a dose-dependent manner. Taken together, these data suggest that DES impairs the morphology and inhibits the proliferation and contractility of gubernaculum testis cells. The experimental model we established and our observations based on this model help provide new insight into the role of DES in the etiology of cryptorchidism.