An immunohistochemical analysis of the effects of androgen receptor knock out on gubernacular differentiation in the mouse

Perera, Nayomi; Szarek, Maciej; Vannitamby, Amanda; Vikraman, Jaya; Huan, Georgina; Durston, Abigale; Hutson, John M.; Li, Ruili

doi:10.1016/j.jpedsurg.2017.11.063

Cited by 5 publications

(6 citation statements)

References 16 publications

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“…The current experiments suggest that androgens do not have a major role in stimulating production of CM or defining muscle fiber size in the rat gubernaculum. These results are consistent with observations that swelling of the gubernaculum still occurs to some extent in mice with spontaneous ( Tfm ) or transgenic (ARKO) Ar inactivation, or in the LE/orl rat, which shows reduced fetal androgen synthesis and action (10, 21, 37). Swelling of the gubernaculum affects its shape, and likely its mechanical properties and function.…”

Section: Discussionsupporting

confidence: 92%

“…We found consistent AR expression in the gubernacular cord, also known as the cranial suspensory ligament (CSL). This finding is consistent with evidence from prior expression studies and from rodent models of AR deficiency in males and androgen exposure in females, which suggest that shortening of the cord is androgen dependent (7, 10, 37, 44–47). INSL3 and DHT regulate many of the same E17 gubernacular transcripts in vitro and both target WNT signaling, but the effects of DHT occur after 24 h, but not 6 h, of stimulation (38).…”

Section: Discussionsupporting

confidence: 91%

“…Loss of AR expression once mesenchymal cells have differentiated into cremaster muscle may explain why conditional Ar deletion in cells that have already undergone myogenic commitment does not interfere with testicular descent (5). Enhanced nuclear AR expression in the tip and cord of the gubernaculum is also consistent with prior studies showing that failure of the cord to shorten and delayed or absent inversion of the gubernaculum occur with loss of AR signaling (36, 37). The present data further indicate that AR is expressed in the gubernaculum during a time frame that overlaps with susceptibility of the rat fetus to antiandrogen-induced cryptorchidism.…”

Section: Discussionsupporting

confidence: 90%

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Regional Variation in Androgen Receptor Expression and Biomechanical Properties May Contribute to Cryptorchidism Susceptibility in the LE/orl Rat

et al. 2018

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Background: The process of testicular descent requires androgen and insulin-like 3, hormones secreted by fetal Leydig cells. Knowledge concerning distinct and common functions of these hormones in regulating development of the fetal gubernaculum remains limited and/or conflicting. The current studies were designed to better define characteristics of androgen receptor (AR) expression, function and regulation, as well as the biomechanical properties of normal and cryptorchid gubernaculum during fetal development.Methods: We studied fetal gubernacula from Long Evans outbred (LE/wt) rats and an inbred (LE/orl) strain with an inherited form of cryptorchidism associated with an AR signaling defect. Gubernacular cells or whole organs obtained from LE/wt and LE/orl fetal gubernacula underwent AR immunostaining and quantitative image analysis. The effects of dihydrotestosterone (DHT) on AR expression, muscle fiber morphology, hyaluronan (HA) levels and glycosaminoglycan (GAG) content were measured in LE/wt gubernacula. Finally, the spatial mechanics of freshly harvested LE/wt and LE/orl fetal gubernacula were compared using micropipette aspiration.Results: AR is expressed in the nucleus of mesenchymal core, tip and cord cells of the embryonic (E) day 17 and 21 fetal gubernaculum, and is enhanced by DHT in primary cultures of gubernacular mesenchymal cells. Enhanced AR expression at the tip was observed in LE/wt but not LE/orl gubernacula. In in vitro studies of whole mount fetal gubernaculum, DHT did not alter muscle fiber morphology, HA content or GAG production. Progressive swelling with reduced cellular density of the LE/wt gubernaculum at E19–21 was associated with increased central stiffness in LE/wt but not in LE/orl fetuses.Conclusions: These data confirm nuclear AR expression in gubernacular mesenchyme with distal enhancement at the tip/cord region in LE/wt but not LE/orl rat fetuses. DHT enhanced cellular AR expression but had no major effects on muscle morphology or matrix composition in the rat fetal gubernaculum in vitro. Regional increased stiffness and decreased cell density between E19 and E21 were observed in LE/wt but not LE/orl fetal gubernacula. Developmental differences in cell-specific AR expression in LE/orl fetal gubernacula may contribute to the dysmorphism and aberrant function that underlies cryptorchidism susceptibility in this strain.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 90%

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Regional Variation in Androgen Receptor Expression and Biomechanical Properties May Contribute to Cryptorchidism Susceptibility in the LE/orl Rat

et al. 2018

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“…At the same time in most mammals a second ligament, the cranial suspensory ligament (CSL), linking the dorsal pole of the testis to the dorsal surface of the body cavity close to the embryonic kidney, needs to dissolve. Studies of androgen receptor knockout (ARKO) and tfm (testicular feminization) male mice, androgen-exposed female mice, and anti-androgen-exposed rats clearly show that the CSL is regulated by androgens and that its persistence leads to cryptorchidism ( 4 – 9 ). Yet CSL regression is thought to facilitate transabdominal rather than transinguinal, descent.…”

Section: Leydig Cell Hormones Work Separately and Together To Bring Amentioning

confidence: 99%

Perspective: A Neuro-Hormonal Systems Approach to Understanding the Complexity of Cryptorchidism Susceptibility

Barthold

Ivell

2018

Front. Endocrinol.

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Nonsyndromic cryptorchidism is a common multifactorial, condition with long-term risks of subfertility and testicular cancer. Revealing the causes of cryptorchidism will likely improve prediction and prevention of adverse outcomes. Herein we provide our current perspective of cryptorchidism complexity in a synthesis of cumulative clinical and translational data generated by ourselves and others. From our recent comparison of genome-wide association study (GWAS) data of cryptorchidism with or without testicular germ cell tumor, we identified RBFOX family genes as candidate susceptibility loci. Notably, RBFOX proteins regulate production of calcitonin gene-related peptide (CGRP), a sensory neuropeptide linked to testicular descent in animal models. We also re-analyzed existing fetal testis transcriptome data from a rat model of inherited cryptorchidism (the LE/orl strain) for enrichment of Leydig cell progenitor genes. The majority are coordinately downregulated, consistent with known reduced testicular testosterone levels in the LE/orl fetus, and similarly suppressed in the gubernaculum. Using qRT-PCR, we found dysregulation of dorsal root ganglia (DRG) sensory transcripts ipsilateral to undescended testes. These data suggest that LE/orl cryptorchidism is associated with altered signaling in possibly related cell types in the testis and gubernaculum as well as DRG. Complementary rat and human studies thus lead us to propose a multi-level, integrated neuro-hormonal model of testicular descent. Variants in genes encoding RBFOX family proteins and/or their transcriptional targets combined with environmental exposures may disrupt this complex pathway to enhance cryptorchidism susceptibility. We believe that a systems approach is necessary to provide further insight into the causes and consequences of cryptorchidism.

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“…Based on the evaluation of cytotoxicity, potency, and cAMP induction efficacy, we selected four compounds for further functional testing in vitro and in vivo. In normal male embryos, it has been described previously that androgen-independent gubernaculum differentiation leads to the processus vaginalis or gubernaculum bulb eversion, which is not observed in female embryos due to the absence of INSL3 expression 50 . Here, we showed that the treatment of pregnant mice with RXFP2 small molecule agonists results in the initiation of the gubernacular eversion in female embryos.…”

Section: Discussionmentioning

confidence: 84%

Discovery of small molecule agonists of the Relaxin Family Peptide Receptor 2

et al. 2022

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The relaxin/insulin-like family peptide receptor 2 (RXFP2) belongs to the family of class A G-protein coupled receptors (GPCRs) and it is the only known target for the insulin-like factor 3 peptide (INSL3). The importance of this ligand-receptor pair in the development of the gubernacular ligament during the transabdominal phase of testicular descent is well established. More recently, RXFP2 has been implicated in maintaining healthy bone formation. In this report, we describe the discovery of a small molecule series of RXFP2 agonists. These compounds are highly potent, efficacious, and selective RXFP2 allosteric agonists that induce gubernacular invagination in mouse embryos, increase mineralization activity in human osteoblasts in vitro, and improve bone trabecular parameters in adult mice. The described RXFP2 agonists are orally bioavailable and display favorable pharmacokinetic properties, which allow for future evaluation of the therapeutic benefits of modulating RXFP2 activation in disease models.

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An immunohistochemical analysis of the effects of androgen receptor knock out on gubernacular differentiation in the mouse

Cited by 5 publications

References 16 publications

Regional Variation in Androgen Receptor Expression and Biomechanical Properties May Contribute to Cryptorchidism Susceptibility in the LE/orl Rat

Regional Variation in Androgen Receptor Expression and Biomechanical Properties May Contribute to Cryptorchidism Susceptibility in the LE/orl Rat

Perspective: A Neuro-Hormonal Systems Approach to Understanding the Complexity of Cryptorchidism Susceptibility

Discovery of small molecule agonists of the Relaxin Family Peptide Receptor 2

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