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John M Hutson - One of the best experts on this subject based on the ideXlab platform.

  • hoxa 11 maintains cell proliferation in the mouse Gubernaculum to facilitate testicular descent
    Journal of Pediatric Surgery, 2013
    Co-Authors: George N Harisis, John M Hutson, Bridget R Southwell, Alfor G Lewis
    Abstract:

    Abstract Introduction The Gubernaculum is a structure vital for guiding testicular descent. The Homeobox gene, Hoxa-11, is involved in patterning embryonic structures and is necessary for gubernacular development, as Hoxa-11 knock-out mice exhibit abnormal gubernacula and undescended testes. We aimed to elucidate how testicular descent fails by examining cell proliferation and androgen receptor (AR) expression in Hoxa-11 KO mice gubernacula. Methods Postnatal day 2 wild type (n=6) and Hoxa-11 KO mice (n=6), were prepared for immunohistochemistry and confocal microscopy using antibodies against androgen receptor, slow skeletal myosin (My32), and Ki67, a marker of cell proliferation. Results The gubernacula of Hoxa-11 KO mice were hypocellular compared with WT. AR was present in the Gubernaculum and abutting inguinal fat pad in both WT and Hoxa-11 KO with no difference in expression. Slow skeletal myosin was present in a clear ‘swirl' in the growth centre of WT animals which was absent in the Hoxa-11 KO mice. Ki67, expressed in the growth centre and cremaster muscle in WT, was greatly decreased in Hoxa-11 KO. Conclusion Hoxa-11 may regulate fibroblast proliferation in the Gubernaculum, as it does in human uterosacral ligaments, allowing formation of the 'growth centre' within the bulb and facilitating myogenesis and elongation to the scrotum. Polymorphisms in Hoxa-11 may contribute to the aetiology of human cryptorchidism.

  • Gubernaculum as icebreaker do matrix metalloproteinases in rodent Gubernaculum and inguinal fat pad permit testicular descent
    Journal of Pediatric Surgery, 2011
    Co-Authors: James Churchill, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, Ruili Li, Adam Balic
    Abstract:

    Abstract Purpose Cryptorchidism is the most common male congenital abnormality. The rodent Gubernaculum steers the testis from abdomen to scrotum postnatally by eversion and migration through the developing inguinal fat pad (IFP). We hypothesize that extracellular matrix remodeling in/around the Gubernaculum is necessary for eversion and migration and is permitted by timed IFP maturation and aimed to examine regional development and matrix metalloproteinase (MMP) content. Methods Embryonic day 19 (E19) and postnatal days 0 and 2 (P0, P2) wild-type Sprague-Dawley rats (n = 10) were prepared for histologic examination (trichrome) and immunohistochemistry (membrane-type MMP-1 [MT1-MMP], MMP2) and analyzed using light/confocal microscopy. Results At E19, IFP contained fibroblasts and immature cells in an extensive collagenous extracellular matrix. Cells in the Gubernaculum base were cytoplasmic-MT1-MMP-positive (inactive). At P0, the Gubernaculum had everted, and adjacent cells were membranous-MT1-MMP-positive (active). At P2, the Gubernaculum was migrating through the IFP, and adjacent cells were membranous-MT1-MMP-positive. Adipocyte maturation began cranially in the IFP and proceeded in a craniocaudal gradient until more uniformly mature at P2. Conclusion The MT1-MMP-positive cells may remodel the Gubernaculum for eversion and provide the collagenolysis necessary for migration, like an icebreaking ship, through the IFP, which matures to permit migration through collagen-rich tissue. Disruption of these processes may cause cryptorchidism.

  • the effect of flutamide on expression of androgen and estrogen receptors in the Gubernaculum and surrounding structures during testicular descent
    Journal of Pediatric Surgery, 2011
    Co-Authors: Tamara R Nation, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, A Balic, Donald F Newgreen
    Abstract:

    Abstract Background/Purpose Inguinoscrotal testicular descent is controlled by androgens between embryonic days E16-19, but androgen receptor (AR) and estrogen receptor (ER) locations are unknown. We aimed to find AR, ER α , and ER β in the Gubernaculum and inguinal fat pad (IFP) in normal rats and after flutamide treatment. Methods Sprague-Dawley timed-mated rats were injected with flutamide (75 mg/kg body weight/5% ethanol + oil) on E16-19 or vehicle alone. Male fetuses or pups (5-10/group) were collected at E16; E19; and postnatal (P) days 0, 2, 4, 8. Sections were prepared for hematoxylin and eosin or immunohistochemistry for AR, ERα, and ERβ. Receptor labeling was quantitated as distinct nuclear labeling/100 μ m 2 in Gubernaculum and IFP. Results There was minimal gubernacular AR-labeling until E19, dramatically increasing postnatally. By contrast, at E16-E19 there was significant IFP AR immunoreactivity suppressed by flutamide ( P Conclusions During the androgen sensitivity window (E16-19), the Gubernaculum contains ERβ but minimal ERα or AR, while the IFP, which is supplied by the genitofemoral nerve, contains abundant AR that are flutamide-sensitive. These results suggest that the IFP could be the site of androgenic action controlling gubernacular development.

  • cremaster muscle myogenesis in the tip of the rat Gubernaculum supports active gubernacular elongation during inguinoscrotal testicular descent
    The Journal of Urology, 2011
    Co-Authors: Nicholas Sanders, Silverton Buraundi, Bridget R Southwell, A Balic, John M Hutson
    Abstract:

    Purpose: Cryptorchidism is a common abnormality and normal testicular descent is controlled by the Gubernaculum. The cremaster may originate from abdominal muscles during gubernacular eversion or alternatively it may develop inside the Gubernaculum. We studied cremaster myogenesis to determine how it develops.Materials and Methods: Coronal sections of the pelvis were prepared from male Sprague-Dawley® rats and from males treated prenatally with the antiandrogen flutamide at embryonic day 19, and postnatal days 10, 19 and 35 after receiving ethical approval. Immunohistochemical stains were prepared for Ki67, Pax-7, myogenin, myosin heavy chain 7, Myh1, Myh2, Myh4, embryonic myosin, and slow and cardiac troponin T. Cell counts of the 1) gubernacular tip, 2) proximal Gubernaculum/cremaster muscle and 3) adjacent abdominal wall are shown as a percent of positive fibers or positive cells per area.Results: Throughout embryonic day 19, and postnatal days 10 and 19 proliferation (Ki67) was maximal at the gubernac...

  • development of the Gubernaculum during testicular descent in the rat
    Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology, 2011
    Co-Authors: Tamara R Nation, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, A Balic, Donald F Newgreen
    Abstract:

    Gubernacular elongation during inguinoscrotal testicular descent and cremaster muscle development remains poorly described in mammals. The role of the genitofemoral nerve (GFN) remains elusive. We performed detailed histological analysis of testicular descent in normal rats to provide a comprehensive anatomical description for molecular studies. Fetuses and neonatal male offspring (5–10 per group) from time-mated Sprague-Dawley dams (embryonic days 15, 16, and 19; postnatal days 0, 2, and 8) were prepared for histology. Immunohistochemistry was performed for nerves (Class III tubulin, Tuj1) and muscle (desmin). At embryonic days 15 and 16, the Gubernaculum and breast bud are adjacent and both supplied by the GFN. By embryonic day 19, the breast bud has regressed and the gubernacular swelling reaction is completed. Postnatally, the gubernacular core regresses, except for a cranial proliferative zone. The cremaster is continuous with internal oblique and transversus abdominis. By postnatal day 2 (P2), the Gubernaculum has everted, locating the proliferative zone caudally and the residual mesenchymal core externally. Eversion creates the processus vaginalis, with the everted Gubernaculum loose in subcutaneous tissue but still remote from the scrotum. By P8, the Gubernaculum has nearly reached the scrotum with fibrous connections attaching the Gubernaculum to the scrotal skin. A direct link between GFN, Gubernaculum, and breast bud suggests that the latter may be involved in gubernacular development. Second, the cremaster muscle is continuous with abdominal wall muscles, but most of its growth occurs in the distal gubernacular tip. Finally, gubernacular eversion at birth brings the cranial proliferative zone to the external distal tip, enabling gubernacular elongation similar to a limb bud. Anat Rec, 2011. © 2011 Wiley-Liss, Inc.

Xuewu Jiang - One of the best experts on this subject based on the ideXlab platform.

  • Impacts of DES on the expressions of related genes in the Gubernaculums testis of newborn mice
    Zhonghua nan ke xue = National journal of andrology, 2020
    Co-Authors: Wei-liao Li, Jianhong Li, Xuan Zhang, Yuan-sheng Du, Xuewu Jiang
    Abstract:

    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. 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. 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. 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.

  • rxfp2 as novel potential biomarker for abnormal differentiation induced by diethylstilbestrol in the Gubernaculum of fetal mice
    American Journal of Translational Research, 2020
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Lian Zheng, Bingna Zhang, Wenfeng Xiao, Jianhong Li
    Abstract:

    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.

  • diethylstilbestrol regulates mouse Gubernaculum testis cell proliferation via plc ca2 creb pathway
    Cell Biochemistry and Function, 2018
    Co-Authors: Xuan Zhang, Xuewu Jiang, Jianhong Li, Hongyan Ping, Shouxin Duan
    Abstract:

    : 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.

  • diethylstilbestrol regulates the expression of lgr8 in mouse Gubernaculum testis cells
    Medical Science Monitor, 2016
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.

  • an insight into insulin like factor 3 regulate its receptor rxfp2 in mouse Gubernaculum testis cells
    International Journal of Clinical and Experimental Pathology, 2015
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.

Jianhong Li - One of the best experts on this subject based on the ideXlab platform.

  • Impacts of DES on the expressions of related genes in the Gubernaculums testis of newborn mice
    Zhonghua nan ke xue = National journal of andrology, 2020
    Co-Authors: Wei-liao Li, Jianhong Li, Xuan Zhang, Yuan-sheng Du, Xuewu Jiang
    Abstract:

    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. 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. 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. 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.

  • rxfp2 as novel potential biomarker for abnormal differentiation induced by diethylstilbestrol in the Gubernaculum of fetal mice
    American Journal of Translational Research, 2020
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Lian Zheng, Bingna Zhang, Wenfeng Xiao, Jianhong Li
    Abstract:

    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.

  • diethylstilbestrol regulates mouse Gubernaculum testis cell proliferation via plc ca2 creb pathway
    Cell Biochemistry and Function, 2018
    Co-Authors: Xuan Zhang, Xuewu Jiang, Jianhong Li, Hongyan Ping, Shouxin Duan
    Abstract:

    : 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.

  • diethylstilbestrol regulates the expression of lgr8 in mouse Gubernaculum testis cells
    Medical Science Monitor, 2016
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.

  • an insight into insulin like factor 3 regulate its receptor rxfp2 in mouse Gubernaculum testis cells
    International Journal of Clinical and Experimental Pathology, 2015
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.

Bridget R Southwell - One of the best experts on this subject based on the ideXlab platform.

  • hoxa 11 maintains cell proliferation in the mouse Gubernaculum to facilitate testicular descent
    Journal of Pediatric Surgery, 2013
    Co-Authors: George N Harisis, John M Hutson, Bridget R Southwell, Alfor G Lewis
    Abstract:

    Abstract Introduction The Gubernaculum is a structure vital for guiding testicular descent. The Homeobox gene, Hoxa-11, is involved in patterning embryonic structures and is necessary for gubernacular development, as Hoxa-11 knock-out mice exhibit abnormal gubernacula and undescended testes. We aimed to elucidate how testicular descent fails by examining cell proliferation and androgen receptor (AR) expression in Hoxa-11 KO mice gubernacula. Methods Postnatal day 2 wild type (n=6) and Hoxa-11 KO mice (n=6), were prepared for immunohistochemistry and confocal microscopy using antibodies against androgen receptor, slow skeletal myosin (My32), and Ki67, a marker of cell proliferation. Results The gubernacula of Hoxa-11 KO mice were hypocellular compared with WT. AR was present in the Gubernaculum and abutting inguinal fat pad in both WT and Hoxa-11 KO with no difference in expression. Slow skeletal myosin was present in a clear ‘swirl' in the growth centre of WT animals which was absent in the Hoxa-11 KO mice. Ki67, expressed in the growth centre and cremaster muscle in WT, was greatly decreased in Hoxa-11 KO. Conclusion Hoxa-11 may regulate fibroblast proliferation in the Gubernaculum, as it does in human uterosacral ligaments, allowing formation of the 'growth centre' within the bulb and facilitating myogenesis and elongation to the scrotum. Polymorphisms in Hoxa-11 may contribute to the aetiology of human cryptorchidism.

  • the effect of flutamide on expression of androgen and estrogen receptors in the Gubernaculum and surrounding structures during testicular descent
    Journal of Pediatric Surgery, 2011
    Co-Authors: Tamara R Nation, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, A Balic, Donald F Newgreen
    Abstract:

    Abstract Background/Purpose Inguinoscrotal testicular descent is controlled by androgens between embryonic days E16-19, but androgen receptor (AR) and estrogen receptor (ER) locations are unknown. We aimed to find AR, ER α , and ER β in the Gubernaculum and inguinal fat pad (IFP) in normal rats and after flutamide treatment. Methods Sprague-Dawley timed-mated rats were injected with flutamide (75 mg/kg body weight/5% ethanol + oil) on E16-19 or vehicle alone. Male fetuses or pups (5-10/group) were collected at E16; E19; and postnatal (P) days 0, 2, 4, 8. Sections were prepared for hematoxylin and eosin or immunohistochemistry for AR, ERα, and ERβ. Receptor labeling was quantitated as distinct nuclear labeling/100 μ m 2 in Gubernaculum and IFP. Results There was minimal gubernacular AR-labeling until E19, dramatically increasing postnatally. By contrast, at E16-E19 there was significant IFP AR immunoreactivity suppressed by flutamide ( P Conclusions During the androgen sensitivity window (E16-19), the Gubernaculum contains ERβ but minimal ERα or AR, while the IFP, which is supplied by the genitofemoral nerve, contains abundant AR that are flutamide-sensitive. These results suggest that the IFP could be the site of androgenic action controlling gubernacular development.

  • Gubernaculum as icebreaker do matrix metalloproteinases in rodent Gubernaculum and inguinal fat pad permit testicular descent
    Journal of Pediatric Surgery, 2011
    Co-Authors: James Churchill, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, Ruili Li, Adam Balic
    Abstract:

    Abstract Purpose Cryptorchidism is the most common male congenital abnormality. The rodent Gubernaculum steers the testis from abdomen to scrotum postnatally by eversion and migration through the developing inguinal fat pad (IFP). We hypothesize that extracellular matrix remodeling in/around the Gubernaculum is necessary for eversion and migration and is permitted by timed IFP maturation and aimed to examine regional development and matrix metalloproteinase (MMP) content. Methods Embryonic day 19 (E19) and postnatal days 0 and 2 (P0, P2) wild-type Sprague-Dawley rats (n = 10) were prepared for histologic examination (trichrome) and immunohistochemistry (membrane-type MMP-1 [MT1-MMP], MMP2) and analyzed using light/confocal microscopy. Results At E19, IFP contained fibroblasts and immature cells in an extensive collagenous extracellular matrix. Cells in the Gubernaculum base were cytoplasmic-MT1-MMP-positive (inactive). At P0, the Gubernaculum had everted, and adjacent cells were membranous-MT1-MMP-positive (active). At P2, the Gubernaculum was migrating through the IFP, and adjacent cells were membranous-MT1-MMP-positive. Adipocyte maturation began cranially in the IFP and proceeded in a craniocaudal gradient until more uniformly mature at P2. Conclusion The MT1-MMP-positive cells may remodel the Gubernaculum for eversion and provide the collagenolysis necessary for migration, like an icebreaking ship, through the IFP, which matures to permit migration through collagen-rich tissue. Disruption of these processes may cause cryptorchidism.

  • cremaster muscle myogenesis in the tip of the rat Gubernaculum supports active gubernacular elongation during inguinoscrotal testicular descent
    The Journal of Urology, 2011
    Co-Authors: Nicholas Sanders, Silverton Buraundi, Bridget R Southwell, A Balic, John M Hutson
    Abstract:

    Purpose: Cryptorchidism is a common abnormality and normal testicular descent is controlled by the Gubernaculum. The cremaster may originate from abdominal muscles during gubernacular eversion or alternatively it may develop inside the Gubernaculum. We studied cremaster myogenesis to determine how it develops.Materials and Methods: Coronal sections of the pelvis were prepared from male Sprague-Dawley® rats and from males treated prenatally with the antiandrogen flutamide at embryonic day 19, and postnatal days 10, 19 and 35 after receiving ethical approval. Immunohistochemical stains were prepared for Ki67, Pax-7, myogenin, myosin heavy chain 7, Myh1, Myh2, Myh4, embryonic myosin, and slow and cardiac troponin T. Cell counts of the 1) gubernacular tip, 2) proximal Gubernaculum/cremaster muscle and 3) adjacent abdominal wall are shown as a percent of positive fibers or positive cells per area.Results: Throughout embryonic day 19, and postnatal days 10 and 19 proliferation (Ki67) was maximal at the gubernac...

  • development of the Gubernaculum during testicular descent in the rat
    Anatomical Record-advances in Integrative Anatomy and Evolutionary Biology, 2011
    Co-Authors: Tamara R Nation, John M Hutson, Pamela J Farmer, Bridget R Southwell, Silverton Buraundi, A Balic, Donald F Newgreen
    Abstract:

    Gubernacular elongation during inguinoscrotal testicular descent and cremaster muscle development remains poorly described in mammals. The role of the genitofemoral nerve (GFN) remains elusive. We performed detailed histological analysis of testicular descent in normal rats to provide a comprehensive anatomical description for molecular studies. Fetuses and neonatal male offspring (5–10 per group) from time-mated Sprague-Dawley dams (embryonic days 15, 16, and 19; postnatal days 0, 2, and 8) were prepared for histology. Immunohistochemistry was performed for nerves (Class III tubulin, Tuj1) and muscle (desmin). At embryonic days 15 and 16, the Gubernaculum and breast bud are adjacent and both supplied by the GFN. By embryonic day 19, the breast bud has regressed and the gubernacular swelling reaction is completed. Postnatally, the gubernacular core regresses, except for a cranial proliferative zone. The cremaster is continuous with internal oblique and transversus abdominis. By postnatal day 2 (P2), the Gubernaculum has everted, locating the proliferative zone caudally and the residual mesenchymal core externally. Eversion creates the processus vaginalis, with the everted Gubernaculum loose in subcutaneous tissue but still remote from the scrotum. By P8, the Gubernaculum has nearly reached the scrotum with fibrous connections attaching the Gubernaculum to the scrotal skin. A direct link between GFN, Gubernaculum, and breast bud suggests that the latter may be involved in gubernacular development. Second, the cremaster muscle is continuous with abdominal wall muscles, but most of its growth occurs in the distal gubernacular tip. Finally, gubernacular eversion at birth brings the cranial proliferative zone to the external distal tip, enabling gubernacular elongation similar to a limb bud. Anat Rec, 2011. © 2011 Wiley-Liss, Inc.

Xuan Zhang - One of the best experts on this subject based on the ideXlab platform.

  • Impacts of DES on the expressions of related genes in the Gubernaculums testis of newborn mice
    Zhonghua nan ke xue = National journal of andrology, 2020
    Co-Authors: Wei-liao Li, Jianhong Li, Xuan Zhang, Yuan-sheng Du, Xuewu Jiang
    Abstract:

    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. 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. 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. 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.

  • rxfp2 as novel potential biomarker for abnormal differentiation induced by diethylstilbestrol in the Gubernaculum of fetal mice
    American Journal of Translational Research, 2020
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Lian Zheng, Bingna Zhang, Wenfeng Xiao, Jianhong Li
    Abstract:

    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.

  • diethylstilbestrol regulates mouse Gubernaculum testis cell proliferation via plc ca2 creb pathway
    Cell Biochemistry and Function, 2018
    Co-Authors: Xuan Zhang, Xuewu Jiang, Jianhong Li, Hongyan Ping, Shouxin Duan
    Abstract:

    : 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.

  • diethylstilbestrol regulates the expression of lgr8 in mouse Gubernaculum testis cells
    Medical Science Monitor, 2016
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.

  • an insight into insulin like factor 3 regulate its receptor rxfp2 in mouse Gubernaculum testis cells
    International Journal of Clinical and Experimental Pathology, 2015
    Co-Authors: Shouxing Duan, Xuewu Jiang, Xuan Zhang, Jianhong Li
    Abstract:

    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.