Estrogen-Regulated Genes in Rat Testes and Their Relationship to Recovery of Spermatogenesis after Irradiation1

Zhou, Wei; Bolden-Tiller, Olga; Shao, Shan; Weng, Connie C.; Shetty, Gunapala; Abuelhija, Mahmoud; Pakarinen, Pirjo; Huhtaniemi, Ilpo; Momin, Amin A.; Wang, Jing; Stivers, David; Liu, Zhilin; Meistrich, Marvin L.

doi:10.1095/biolreprod.111.091611

Cited by 17 publications

(10 citation statements)

References 61 publications

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“…The expression of Insl3 in MA-10 Leydig cells is similar to what has been reported in primary Leydig cell cultures and in rodents. For instance, Insl3 expression is known to be repressed by estradiol in vivo in mice and rats (Nef et al 2000, Cederroth et al 2007, Strauss et al 2009, Zhou et al 2011) and this repression was also observed in MA-10 Leydig cells ). Insl3 expression is upregulated by androgens in vivo (Zhou et al 2010) as well as in primary Leydig cells and in MA-10 Leydig cells (Laguë & Tremblay 2008).…”

Section: Cell Culture Transfections and Reporter Assaysmentioning

confidence: 94%

The INSL3 gene is a direct target for the orphan nuclear receptor, COUP-TFII, in Leydig cells

Mendoza-Villarroel¹,

Di-Luoffo²,

Camiré³

et al. 2014

Journal of Molecular Endocrinology

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Insulin-like 3 (INSL3), a hormone produced by Leydig cells, regulates testicular descent during foetal life and bone metabolism in adults. Despite its importance, little is known about the molecular mechanisms controlling INSL3 expression. Reduced Insl3 mRNA levels were reported in the testis of mice deficient for chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII), an orphan nuclear receptor known to play critical roles in cell differentiation and lineage determination in several tissues. Although COUP-TFII-deficient mice had Leydig cell dysfunction and impaired fertility, it remained unknown whether Insl3 expression was directly regulated by COUP-TFII. In this study, we observed a significant decrease in Insl3 mRNA levels in MA-10 Leydig cells depleted of COUP-TFII. Furthermore, a K1087 bp mouse Insl3 promoter was activated fourfold by COUP-TFII in MA-10 Leydig cells. Using 5 0 progressive deletions, the COUP-TFII-responsive element was located between K186 and K79 bp, a region containing previously uncharacterised direct repeat 0-like (DR0-like) and DR3 elements. The recruitment and direct binding of COUP-TFII to the DR0-like element were confirmed by chromatin immunoprecipitation and DNA precipitation assay respectively. Mutation of the DR0-like element, which prevented COUP-TFII binding, significantly decreased COUP-TFII-mediated activation of the K1087 bp Insl3 reporter in CV-1 fibroblast cells but not in MA-10 Leydig cells. Finally, we found that COUP-TFII cooperates with the nuclear receptor steroidogenic factor 1 (SF1) to further enhance Insl3 promoter activity. Our results identify Insl3 as a target for COUP-TFII in Leydig cells and revealed that COUP-TFII acts through protein-protein interactions with other DNA-bound transcription factors, including SF1, to activate Insl3 transcription in these cells.

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Section: Cell Culture Transfections and Reporter Assaysmentioning

confidence: 94%

The INSL3 gene is a direct target for the orphan nuclear receptor, COUP-TFII, in Leydig cells

Mendoza-Villarroel¹,

Di-Luoffo²,

Camiré³

et al. 2014

Journal of Molecular Endocrinology

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“…Although type A spermatogonial proliferation is initiated within 2 weeks after hormone suppression (Zhou et al , 2011), 6 weeks are required for a robust stimulation of differentiation of surviving type A spermatogonia to the B spermatogonial and spermatocyte stages (Porter et al , 2006; Shuttlesworth et al , 2000). The reasons for this delay are also unknown.…”

Section: Introductionmentioning

confidence: 99%

“…Analyses of microarray data indicated that most of the genes that were upregulated after irradiation were dependent on T were expressed in Leydig cells (LCs) (Zhou et al , 2011; Zhou et al , 2010). Hence in the present study, we studied the LCs as potential effectors of the T-induced inhibition of spermatogonial differentiation.…”

Section: Introductionmentioning

confidence: 99%

Leydig cells contribute to the inhibition of spermatogonial differentiation after irradiation of the rat

et al. 2016

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SUMMARY Irradiation with 6 Gy produces a complete block of spermatogonial differentiation in LBNF1 rats that would be permanent without treatment. Subsequent suppression of gonadotropins and testosterone (T) restores differentiation to the spermatocyte stage; however this process requires 6 weeks. We evaluated the role of Leydig cells (LCs) in maintenance of the block in spermatogonial differentiation after radiation exposure by specifically eliminating functional LCs with ethane dimethane sulfonate (EDS). EDS (but not another alkylating agent), given at 10 weeks after irradiation, induced spermatogonial differentiation in 24% of seminiferous tubules 2 weeks later. However differentiation became blocked again at 4 weeks as LCs recovered. When EDS was followed by treatment with GnRH antagonist and flutamide, sustained spermatogonial differentiation was induced in >70% of tubules within 2 weeks. When EDS was followed by GnRH antagonist plus exogenous T, which also inhibits LC recovery but restores follicle stimulating hormone (FSH) levels, the spermatogonial differentiation was again rapid but transient. These results confirm that the factors that block spermatogonial differentiation are indirectly regulated by T and probably FSH and that adult and possibly immature LCs contribute to the production of such inhibitory factors. We tested whether insulin-like 3 (INSL3), a LC-produced protein whose expression correlated with the block in spermatogonial differentiation, was indeed responsible for the block by injecting synthetic INSL3 into the testis and knocking down its expression in vivo with siRNA. Neither treatment had any effect on spermatogonial differentiation. The Leydig cell products that contribute to the inhibition of spermatogonial differentiation in irradiated rats remain to be elucidated.

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“…Rarely, larger protrusions into A 1 spermatogonia that were only attached to the basement membrane at the protrusion were observed and may be a first sign the death of the spermatogonia. Based on observations that the genes most strongly increased after irradiation and downregulated by androgen suppression in the somatic cells of rat testes were in the Leydig cells (Zhou et al 2010(Zhou et al , 2011, we further investigated the role of Leydig cells in the spermatogonial differentiation block by injecting EDS into irradiated rats at the beginning of the hormone suppression treatment. The loss of Leydig cells produced by EDS, in addition to hormonal suppression, produced rapid recovery of spermatogenesis resulting in almost the same numbers of differentiating spermatogonia within 1 week as produced by 2 weeks of hormone suppression alone.…”

Section: Discussionmentioning

confidence: 99%

“…Gene expression profiles in the irradiated rats and irradiated rats with hormone suppression have also been analyzed (Zhou et al 2010(Zhou et al , 2011. In this study, we have extended the characterization of the kinetics and molecular events of spermatogonial differentiation in rats during spermatogenic recovery after testis irradiation and hormone suppression with a GNRH antagonist (GNRH-ant), using morphological (light and transmission electron microscopy) and stereological approaches and molecular markers (e.g., KIT).…”

Section: Introductionmentioning

confidence: 99%

Spermatogonial behavior in rats during radiation-induced arrest and recovery after hormone suppression

Albuquerque¹,

Almeida²,

Weng³

et al. 2013

Reproduction

Self Cite

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Ionizing radiation has been shown to arrest spermatogenesis despite the presence of surviving stem spermatogonia, by blocking their differentiation. This block is a result of damage to the somatic environment and is reversed when gonadotropins and testosterone are suppressed, but the mechanisms are still unknown. We examined spermatogonial differentiation and Sertoli cell factors that regulate spermatogonia after irradiation, during hormone suppression, and after hormone suppression combined with Leydig cell elimination with ethane dimethane sulfonate. These results showed that the numbers and cytoplasmic structure of Sertoli cells are unaffected by irradiation, only a few type A undifferentiated (A und ) spermatogonia and even fewer type A 1 spermatogonia remained, and immunohistochemical analysis showed that Sertoli cells still produced KIT ligand (KITLG) and glial cell line-derived neurotrophic factor (GDNF). Some of these cells expressed KITreceptor, demonstrating that the failure of differentiation was not a result of the absence of the KIT system. Hormone suppression resulted in an increase in A und spermatogonia within 3 days, a gradual increase in KIT-positive spermatogonia, and differentiation mainly to A 3 spermatogonia after 2 weeks. KITL (KITLG) protein expression did not change after hormone suppression, indicating that it is not a factor in the stimulation. However, GDNF increased steadily after hormone suppression, which was unexpected since GDNF is supposed to promote stem spermatogonial self-renewal and not differentiation. We conclude that the primary cause of the block in spermatogonial development is not due to Sertoli cell factors such (KITL\GDNF) or the KIT receptor. As elimination of Leydig cells in addition to hormone suppression resulted in differentiation to the A 3 stage within 1 week, Leydig cell factors were not necessary for spermatogonial differentiation.

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Estrogen-Regulated Genes in Rat Testes and Their Relationship to Recovery of Spermatogenesis after Irradiation1

Cited by 17 publications

References 61 publications

The INSL3 gene is a direct target for the orphan nuclear receptor, COUP-TFII, in Leydig cells

The INSL3 gene is a direct target for the orphan nuclear receptor, COUP-TFII, in Leydig cells

Leydig cells contribute to the inhibition of spermatogonial differentiation after irradiation of the rat

Spermatogonial behavior in rats during radiation-induced arrest and recovery after hormone suppression

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