Testicular descent revisited: a micro-computed tomography study in fetal rats.

PURPOSE: There is a long history of research dealing with the embryology of the testicular descent. However, important aspects like the role of the gubernaculum and the development of the processus vaginalis peritonei are not understood. Micro-computed tomography (µCT) is an established tool for anatomical studies in rodents. Our study applied µCT imaging to visualize the testicular descent in rats and focused on the role of the gubernacular bulb and the development of the processus vaginalis peritonei. METHODS: Rats from embryonic day 15 (ED15) to ED21 and newborns (N0) were fixed and dried using the "critical point" technique. We ran a SkyScan® µCT system and scans were analyzed for gender-specific differentiation of the genital ridge and used for 3D visualization of relevant anatomic structures. RESULTS: µCT imaging confirmed the intraperitoneal location of the testicles from ED15 to N0. The components of the inner genital moved closer together while the intestinal volume expanded. The gubernacular bulb seemed to be involved in the formation of the processus vaginalis peritonei. CONCLUSION: Here, we utilized µCT imaging to visualize the testicular descent in the rat. Imaging provides new morphologic aspects on the development of the processus vaginalis peritonei.

Testicular Torsion: Preliminary Results of In Vitro Cell Stimulation Using Chorionic Gonadotropin.

Testicular torsion is a pathology that occurs in young males generally before the age of 25. Despite surgery representing the only effective approach, there is still a need to identify a marker that can predict whether a preserved testicle will be functional. In addition, no therapeutic approach is currently considered in the post-operative phase. Through an approach based on the in vitro culture of a tissue strictly linked to the testicle, the gubernaculum, we defined the healthy state of the organ and the possible responsiveness to a therapy used in the andrology field, chorionic gonadotropin (hCG). Firstly, we optimized a protocol to obtain viable cells starting from a small piece of gubernacular tissue harvested during surgery with the aim to amplify cells in vitro. Intriguingly, only for a patient whose testicle had been removed during surgery due to an excessive necrotic area, gubernacular cells were not able to grow in culture. These data support the possibility of exploiting the gubernaculum to evaluate the healthy state of the testicle. Then, as we demonstrate that gubernacular cells express a luteinizing hormone receptor, to which hCG is specific, we analyzed the cellular response to hCG treatment on in vitro cultured cells derived from patients affected by testicular torsion. Our study opens the way for the possibility of evaluating testicle wellbeing after derotation through in vitro culture of a small piece of gubernaculum together with predicting the response to the treatment with hCG, which can have a positive effect on cell proliferation and vascularization.

Testicular blood supply and growth in children with high cryptorchidism treated with gubernaculum preserving vs. cutting laparoscopic orchiopexy: A pilot trial.

INTRODUCTION: It is still controversial whether the frenum of testis should be retained in laparoscopic testicular fixation. MATERIALS AND METHODS: It is a prospective experiment to study testicular preservation on testicular growth in children with high cryptorchidism. RESULTS: From January 2018 to June 2020, 120 children with high cryptorchidism in Inner Mongolia Maternal and Child Health Care Hospital and The Fourth Hospital of Baotou were randomly divided into retention group (60 cases in group P, aged 1-3 years, average 1.85 ± 0.58) and cutting group (60 cases in group C, aged 1-3 years, average 1.75 ± 0.66) (P = 0.52). PSV, EDV, RI and testicular volume were measured by color Doppler ultrasound before operation and 1,3,6,12 months after operation. No, there was significant difference between the two groups (P > 0.05). CONCLUSIONS: There is no significant difference in testicular blood supply and growth between laparoscopic testicular fixation and frenulum amputation in children with high cryptorchidism.

Inguinoscrotal stage of testicular descent: analysis in 217 human fetuses.

BACKGROUND: Testicular descent is a complex process of relevant importance for the comprehension of cryptorchidism. Studies about the inguinoscrotal stage of testicular migration correlating the testicular position with fetal parameters are rare in the literature. Our hypothesis is that the transition of the testis by the inguinal canal is a very fast process and occurs at the end of the third gestational trimester. AIM: To analyze the inguinoscrotal stage of testicular descent during the human fetal period. STUDY DESIGN: This is an observational study in human fetuses. We studied 217 human male fetuses ranging from 10 to 35 weeks postconception (WPC). The fetuses were well preserved and evaluated regarding crown-rump length (CRL) and body weight before dissection. Testicular position was classified as: a) abdominal, when the testis was proximal to the internal ring; b) inguinal, when it was found between the internal and external inguinal rings; and c) scrotal, when being inside the scrotum. Means were statistically compared using the unpaired t-test (P < 0.05). RESULTS: Of the 434 testes, 329 (75.8%) were abdominal, 48 (11.05%) were inguinal, and 57 (13.13%) were scrotal. The first case of inguinal testis was observed in a fetus with 17 WPC, 203 g, and 15 cm CRL. The 48 inguinal testes were observed in 29 fetuses (13.36%), aged between 17 and 29 WPC, weighing between 203 and 1220 g, and with CRL between 15 and 27.5 cm. Of the 29 fetuses with inguinal testes, 23 (79.32%) were aged between 20 and 26 WPC; 3 (10.34%) had 17 WPC; and 3 (10.34%) had more than 26 WPC. All the fetuses with more than 30 WPC had testes in the scrotum. We did not observe fetuses less than 17 WPC with the testes in the canal. DISCUSSION: We found that all fetuses with more than 30 WPC had the testes located in the scrotum and that in fetuses between 17 WPC and 25 WPC, 38 testes (8.7% of the sample) were situated in the canal and only 1 testis in a fetus with 25 WPC was located in the scrotum, showing that the second stage of testicular migration through the inguinal canal rarely occurs before the 20th WPC. CONCLUSION: The inguinoscrotal stage of testicular descent is a fast process, observed only in 13% of the fetuses and occurring with more intensity between 20 and 26 WPC.

Anatomy and histology of the scrotal ligament in adults: inconsistency and variability of the gubernaculum testis.

PURPOSE: The anatomy of gubernaculum testis (GT) is often discussed; however, the postnatal anatomy of the GT or scrotal ligament (SL) is rarely described. Hence, we performed an anatomical and histological study to analyze histologically the structures between testis and scrotum. METHODS: We performed anatomical dissections on 25 human fresh cadavers' testes. Each testis was removed with its envelopes and macroscopically analyzed. Then samples were included for histological study. Finally, they were analyzed under microscope, looking for attachments between testis, epididymis and scrotal envelopes. RESULTS: The absence of proximal and distal attachment was found in 56.0% of cases. Looking at the proximal attachment of the SL, the main one found is the epididymal attachment (28.0%), whereas no cases of testis attachment was found. Distally, there are more variations with scrotal attachment (12%) and cremaster attachment (12.0%). We found a significant prevalence of multiple adherences in 16.0% of cases too. Finally, in 15 cases (57.7%) an attachment is present between testis and epididymis, as it is commonly described. CONCLUSIONS: In the majority of cases there is no attachment of the lower pole of the testis and epididymis and these structures remain free. So it seems that the SL disappears with aging. Moreover, there is not only one kind of ligamentous attachment, but a high variability of attachments at the lower pole of the testiculo-epididymal structure. When it exists, this structure is never a real ligament and it seems more appropriate to use the term "attachments".

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.

Impacts of DES on the expressions of related genes in the gubernaculums testis of newborn mice / 中华男科学杂志

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.

Is there a trans-abdominal testicular descent during the second gestational trimester? Study in human fetuses between 13 and 23 weeks post conception

ABSTRACT Objectives To confirm if a real inner descend of testis occurs, correlating the testicular position with fetal parameters and analyzing the position of the testes relative to the internal ring. Material and Methods Twenty nine human fetuses between 13 and 23 weeks post conception (WPC) were studied. The fetuses were carefully dissected with the aid of a stereoscopic lens with 16/25X magnification and testicular position observed. With the aid of a digital pachymeter the distance between the lower pole of the kidney and the upper extremity of the testis (DK-T) was measured to show the position of the testis. During the dissection we also indicated the position of the testes relative to the internal ring. Means were statistically compared using simple linear regression and the paired T-test. Results The 58 testes had abdominal position. The DK-T in the right side measured between 0.17 and 1.82cm (mean=0.79cm) and in the left side it was between 0.12 and 1.84cm (mean=0.87cm), without statistically differences (p=0.0557). The linear regression analysis indicated that DK-T in both sides correlated significantly and positively with fetal age. All fetuses with more than 20 WPC, heavier than 350g and with CRL over 22cm had a greater distance than the average DK-T. We xobserved that the 58 testis remains adjacent to the internal ring throughout the period studied. Conclusions The testes remains adjacent to the internal ring throughout the period studied, indicating that there is no real trans-abdominal testicular descent during the second gestational trimester.

Aquaporin water channels in the canine gubernaculum testis.

The jelly-like gubernaculum testis (GT) is a hydrated structure consisting of a concentric sheath of dense connective tissue around a loose mesenchymal core, with two cords of skeletal muscle cells asymmetrically placed alongside. Expansion of the GT occurs during the transabdominal phase of testicular descent, linked to cell proliferation together with modifications of the hydric content of the organ. The aim of this study was to detect immunohistochemically the presence of aquaporins (AQPs), integral membrane proteins permitting passive transcellular water movement, in the canine GTs. Samples (n=15) were obtained from pregnancies of 9 medium sized bitches and dissected from healthy fetuses. Five fetuses were aged 35-45 days of gestation, 10 fetuses from 46 days of gestation to delivery, thus offering us the opportunity to study the progressive maturation of the gubernacula. The presence of AQP3, 4, 7, 8 and -9 was assessed in the muscular components of the GT, some of them (AQP3, AQP4, AQP7) with increasing intensity through the second half of pregnancy up to term. AQP1 was localized in the capillary and venous endothelia in the younger fetuses, also in the artery adventitia and in the nerve perineurium in progressively older fetuses. These data demonstrate the potential importance and contribution of AQP-mediated water flux in hydration and volume modification of the growing GT in a canine model.

Anatomical Study on the Descent of Testis in Prenatal Goat (Capra hircus) / Estudio Anatómico del Descenso Testicular Prenatal en la Cabra (Capra hircus)

A study was conducted on 70 embryo/ foetii of goats to observe the descent of testis. These foetii were grouped into I (0-30 days), II (31-60 days), III (61-90 days), IV (91-120days) and V (121 days up to term, having 14 embryos/ foetii in each group. The genital ridge was observed at 1.2cm CRL (23 day) on the ventromedial aspect of mesonephros. At 42nd day, the genital ridge developed into cylindrical structure, the testis, located in the middle of metanephros. The abdominal migration was completed upto 88th day. The testes were situated in the inguinal canal from 89th to 95th day. From 90th day onward, scrotal migration had been observed, however the testis did not reach the base of the scrotum up to term. During the descent of testis, a peritoneal fold detached from the caudal end of the gonad and extended up-to the abdominal floor on 42nd day, known as gubernaculum. In the initial stages of pregnancy it was thin, whitish, jelly like. On 70th day it was enveloped by processus vaginalis. In group V, it became hardened, longer and cord like.

Para observar el descenso de los testículos se realizó un estudio en 70 embriones/fetos de cabra. Estos fetos se agruparon en 5 grupos: I (0-30 días), II (31-60 días), III (61-90 días), IV (91 120 días) y V (121 días hasta término, con 14 embriones / fetos en cada grupo. La cresta genital se observó a 1,2cm CRL (23 días) en la cara ventromedial del mesonefros. En el día 42 día, la cresta genital se convirtió en estructura cilíndrica y los testículos se encontraban en el centro del metanefros. La migración abdominal se completó en 88 días. Los testículos están situados en el canal inguinal entre los 89 y 95 días. Del día 90 en adelante, se observó la migración escrotal, sin embargo, el testículo no llegó a la base del escroto hasta el término del desarrollo. Durante el descenso de los testículos, a los 42 días, un pliegue peritoneal (gubernaculum testis) separado del extremo caudal de la gónada, se extendió hacia el suelo-abdominal. En las etapas iniciales de la preñez la gónada era delgada, blanquecina, gelatinosa. En el día 70 el testículo fue envuelto por el proceso vaginal. En el grupo V, el testículo se había endurecido y alargado asemejándose a una cuerda.

Effect of antiandrogens upon the morphological development of gubernaculum and the testicular descent in rats / 대한비뇨기과학회지

We evaluated the effect of antiandrogens on both fetal and adult rats, noting especially the morphological changes in the gubernaculum and testicular descent. Two antiandrogens, flutamide and 5a-reductase inhibitor 4-methyl-4-aza-5-pregnane-3-one-20[s] carboxylate (4-MAPC) produce gross and histological changes in the gubernaculum, as well as having an effect on testicular descent. While the gubernacula of controls were like full water-drop at day 20 of gestation. the gubernacula of fetuses treated with flutamide were thin and elliptical and the gubernacula of fetuses treated with 4-MAPC were thin and elliptical or like small water-drop. The position of the gubernaculum was near the bladder neck and lower border of pelvic cavity, regardless of the antiandrogen treatment. But, the degree of invagination of gubernaculum into abdominal wall tended to be less in fetuses treated with either antiandrogen. The muscular layer of gubernaculum treated with either anti-androgen tended to be thinner than controls. In adult rats treated from embryo to adult with flutamide, atrophy of the sex accessory glands and hypospadias were occurred and the scrotums were bifid or undeveloped on one side which the testis remained in the abdomen. We feel that flutamide and 4-MAPC cannot interfere with trans-abdominal testicular descent, but can induce the morphological changes in gubernaculum with failure of the trans-inguinal testicular descent. Also, it would be suggested that the gubernaculum has not Wolffian duct like testosterone dependency shown in differentiation of epididymis, vas deferens and seminal vesicle, because of the presence of the effect of both flutamide and 4-MAPC on gubernaculum.

Genitofemoral nerve and undescended testis / 대한비뇨기과학회지

The gubernaculum testis, which has an important role to play in transinguinal testicular descent, is richly by the genitofemoral nerve through its scrotal attachment. In neonatal rats the genitofemoral nerve overlying the psoas muscle was divided before transinguinal descent would normally occur, and the effect of this procedure on subsequent testicular descent was observed and compared with the effect of division of psoas muscle and gubernaculectomy. Selective division of the genitofemoral nerve could not cause undescended testis and testicular atrophy which was caused by the division of the psoas muscle or gubernaculectomy. However denervation of the gubernaculum shortened the length of the spermatic cord and vas deference. This effect tended to be obvious in group or earlier neonatal denervation. The significance of this finding in the rat is that an intact genitofemoral nerve is an essential prerequisite for normal transinguinal descent perhaps by allowing the gubernaculum to evert and to differentiate.