Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells

Zhang, Qiao-Xia; Zhang, Xiaoyan; Zhang, Zhenming; Lu, Wei; Liu, Ling; Li, Gang; Cai, Zhiming; Gui, Yaoting; Chang, Chawnshang

doi:10.1038/aja.2011.94

Cited by 30 publications

(20 citation statements)

References 30 publications

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“…The key function of testosterone is to activate spermatogenesis by acting on Sertoli cells, which provide nutritional as well as morphogenetic support for germ cells during Circadian alteration of reproduction exposed to RF 129 ! Informa Healthcare USA, Inc. Chronobiol Int Downloaded from informahealthcare.com by Dalhousie University on 12/26/14 spermatogenesis via androgen receptor (Griswold, 1998;Zhang et al, 2012). g-GT and ACP are two marker enzymes of Sertoli cells and their activities are representative of Sertoli cell function.…”

Section: Discussionmentioning

confidence: 99%

Circadian alterations of reproductive functional markers in male rats exposed to 1800 MHz radiofrequency field

Qin

Zhang²,

Cao

et al. 2013

Chronobiology International

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In this study, we explored the circadian effects of daily radiofrequency field (RF) exposure on reproductive functional markers in adult male Sprague-Dawley rats. Animals in circadian rhythm (as indicated by melatonin measurements), were divided into several groups and exposed to 1800 MHz RF at 205 μw/cm(2) power density (specific absorption rate 0.0405 W/kg) for 2 h/day for 32 days at different zeitgeber time (ZT) points, namely, ZT0, ZT4, ZT8, ZT12, ZT16 and ZT20. Sham-exposed animals were used as controls in the study. From each rat, testicular and epididymis tissues were collected and assessed for testosterone levels, daily sperm production and sperm motility, testis marker enzymes γ-GT and ACP, cytochrome P450 side-chain cleavage (p450cc) mRNA expression, and steroidogenic acute regulatory protein (StAR) mRNA expression. Via these measurements, we confirmed the existence of circadian rhythms in sham-exposed animals. However, rats exposed to RF exhibited a disruption of circadian rhythms, decreased testosterone levels, lower daily sperm production and sperm motility, down-regulated activity of γ-GT and ACP, as well as altered mRNA expression of cytochrome P450 and StAR. All of these observations were more pronounced when rats were exposed to RF at ZT0. Thus, our findings indicate potential adverse effects of RF exposure on male reproductive functional markers, in terms of both the daily overall levels as well as the circadian rhythmicity.

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Section: Discussionmentioning

confidence: 99%

Circadian alterations of reproductive functional markers in male rats exposed to 1800 MHz radiofrequency field

Qin

Zhang²,

Cao

et al. 2013

Chronobiology International

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“…Briefly, TM4 cells purchased from American Tissue Culture Collection (Manassas, VA, USA) were seeded in a 96-well plate (7.0 9 10 3 cells) and transiently transfected with a pcDNA3.1 plasmid that expresses the mouse androgen receptor sequence (pcDNA-AR, kindly donated by Dr. Chawnshang Zhang from Rochester University, USA), with a commercial plasmid that expresses luciferase gene, whose promoter sequence possesses tandem androgen response elements (AREs, Cignal Pathway Reporter kit; Qiagen) and with a pGEN2 plasmid that contains b-galactosidase sequence (pGEN2-bgal, kindly donated by Dr. Cecilia Johnson from University of Chile) following a protocol previously described by Zhang et al (2012). Briefly, TM4 cells purchased from American Tissue Culture Collection (Manassas, VA, USA) were seeded in a 96-well plate (7.0 9 10 3 cells) and transiently transfected with a pcDNA3.1 plasmid that expresses the mouse androgen receptor sequence (pcDNA-AR, kindly donated by Dr. Chawnshang Zhang from Rochester University, USA), with a commercial plasmid that expresses luciferase gene, whose promoter sequence possesses tandem androgen response elements (AREs, Cignal Pathway Reporter kit; Qiagen) and with a pGEN2 plasmid that contains b-galactosidase sequence (pGEN2-bgal, kindly donated by Dr. Cecilia Johnson from University of Chile) following a protocol previously described by Zhang et al (2012).…”

Section: Evaluation Of Testosterone-induced Transcriptional Activity mentioning

confidence: 99%

Disturbed testicular expression of the estrogen‐metabolizing enzymes CYP1A1 and COMT in infertile men with primary spermatogenic failure: possible negative implications on Sertoli cells

et al. 2017

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Estradiol (E ) is normally metabolized to hydroxyestradiols and methoxyestradiols by CYP1A1, CYP1B1 and COMT. However, an altered production of these metabolites by a disturbed expression of these enzymes is associated with reproductive and non-reproductive pathologies. In vitro studies suggest that increased hydroxyestradiols and methoxyestradiols intratesticular generation is related to male infertility, but no studies have explored whether infertile men have a disturbed testicular expression of the enzymes that generate these E metabolites. The aim of this study was to assess CYP1A1, CYP1B1 and COMT testicular expression at mRNA and protein level in men with spermatogenic impairment. Seventeen men with primary spermatogenic failure (13 with Sertoli cell-only syndrome and four with maturation arrest) and nine controls with normal spermatogenesis were subjected to testicular biopsy. mRNA was quantified using real-time RT-PCR and protein expression was evaluated using western blot and immunohistochemistry followed by integrated optic density analysis. Besides, the effects of hydroxyestradiols and methoxyestradiols on testosterone-induced transcriptional activity were evaluated in TM4 cells using a luciferase reporter assay system. Our results show that patients with Sertoli cell-only syndrome had significantly elevated COMT expression at the mRNA level, higher COMT immunoreactivity in their seminiferous tubules and increased protein expression of the soluble COMT isoform (S-COMT), whereas patients with maturation arrest had significantly elevated CYP1A1 mRNA levels and higher CYP1A1 immunoreactivity in interstitial space. Finally, 2-hydroxyestradiol decreased testosterone-induced transcriptional activity in Sertoli cells in vitro. In conclusion, male infertility is related to disturbed testicular expression of the enzymes responsible for producing hydroxyestradiols and/or methoxyestradiols. If these changes are related with increased intratesticular hydroxyestradiols and methoxyestradiols concentrations, they could elicit an impaired Sertoli cell function. Our results suggest CYP1A1 and COMT as new potential targets in treating male infertility.

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“…TM4 cells were grown in a 1:1 mixture of Ham's F-12 medium and Dulbecco's modified Eagle's medium (DMEM) supplemented with 1-2 g/l sodium bicarbonate and 15 mM HEPES, 5% horse serum, and 2.5% fetal calf serum (FCS) and incubated in a humidified atmosphere of 10% CO 2 and 90% air at 37°C. Since its generation, this cell line has been used in numerous studies with results similar to those obtained using primary cultures of Sertoli cells (45,49,71).…”

Section: Methodsmentioning

confidence: 87%

Sertoli-secreted FGF-2 induces PFKFB4 isozyme expression in mouse spermatogenic cells by activation of the MEK/ERK/CREB pathway

Gómez

Manzano

Figueras

et al. 2012

American Journal of Physiology-Endocrinology and Metabolism

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In this study, we investigated how the hormonal regulation of spermatogenesis modulates 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) isozyme expression in two mouse spermatogenic cell lines, GC-1 spg and GC-2 spd (ts). For this purpose, TM4 Sertoli cells were used to obtain conditioned medium that was treated or not with dihydrotestosterone for 2 days [dihydrotestosterone conditioned medium (TCM) and basal conditioned medium (BCM), respectively]. We observed an increase in the expression of PFKFB4 along with a decrease in PFKFB3 in spermatogenic cell lines treated with TCM. These effects were inhibited by the antiandrogen drug flutamide and by heat-inactivated TCM, indicating the protein nature of the TCM mediator and its dependence on Sertoli cell stimulation by dihydrotestosterone. In addition, adult rat testes treated with the GnRH antagonist Degarelix exhibited a reduction in the expression of PFKFB4 in germ cells. Addition of exogenous FGF-2 mimicked the changes in the Pfkfb gene expression, whereas neutralizing antibodies against FGF-2 abolished them. Interestingly, similar effects on Pfkfb gene expression were observed using different MAPK inhibitors (U-0126, PD-98059, and H-89). Luciferase analysis of Pfkfb4 promoter constructs demonstrated that a putative CRE-binding sequence located at Ϫ1,463 relative to the transcription start site is required to control Pfkfb4 gene expression after TCM treatment. Pulldown assays showed the binding of the CREB transcription factor to this site. Altogether, these results show how the paracrine regulation orchestrated by Sertoli cells in response to testosterone controls glycolysis in germ cells.fibroblast growth factor; 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase; mitogen-activated protein kinase kinase; extracellular signal-regulated kinase; cAMP response element-binding protein; fructose-2,6-bisphosphate; glycolysis; spermatogenesis; sperm production

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Identification of testosterone-/androgen receptor-regulated genes in mouse Sertoli cells

Cited by 30 publications

References 30 publications

Circadian alterations of reproductive functional markers in male rats exposed to 1800 MHz radiofrequency field

Circadian alterations of reproductive functional markers in male rats exposed to 1800 MHz radiofrequency field

Disturbed testicular expression of the estrogen‐metabolizing enzymes CYP1A1 and COMT in infertile men with primary spermatogenic failure: possible negative implications on Sertoli cells

Sertoli-secreted FGF-2 induces PFKFB4 isozyme expression in mouse spermatogenic cells by activation of the MEK/ERK/CREB pathway

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