Estradiol and 1α,25(OH)2 vitamin D3 share plasma membrane downstream signal transduction through calcium influx and genomic activation in immature rat testis

Zanatta, Ana Paula; Gonçalves, Renata; Ourique, Fabiana; Pedrosa, Rozangela Curi; Zanatta, Leïla; Bouraïma-Lelong, Hélène; Delalande, Christelle; Silva, Fátima Regina Mena Barreto

doi:10.1016/j.theriogenology.2021.05.030

“…Additionally, estrogen signaling was impaired in organotypic cultures, with a low expression of the estrogen-synthesizing enzyme aromatase, of estrogen receptors and of the estrogen target gene Faah ( Figure 7 ). Since the expression of Cyp19a1 , Esr1, and Esr2 (encoding aromatase, ERα, and ERβ, respectively) can be downregulated by estradiol in the rat testis ( Zanatta et al, 2021 ; Genissel et al, 2001 ), we hypothesize that the transcription of these genes may be negatively controlled by elevated estradiol levels in our tissue cultures. Faah , whose promoter activity engages ERβ and the histone demethylase LSD1, is a direct target gene of estrogens in Sertoli cells ( Grimaldi et al, 2012 ).…”

Section: Discussionmentioning

confidence: 99%

Steroidogenesis and androgen/estrogen signaling pathways are altered in in vitro matured testicular tissues of prepubertal mice

Moutard

¹

,

Goudin

²

,

Jaeger

³

et al. 2023

eLife

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Cancer treatments such as chemotherapy can have gonadotoxic effects. In order to preserve and restore the fertility of prepubertal patients with cancer, testicular biopsies are frozen and could theoretically be later matured in vitro to produce spermatozoa for assisted reproductive technology. A complete in vitro spermatogenesis has been obtained from prepubertal testicular tissue in the mouse model, although the sperm yield was low. Since steroid hormones play an essential role in spermatogenesis, it appears necessary to ensure that their synthesis and mechanisms of action are not altered in in vitro cultured tissues. The aim of this study was therefore to investigate steroidogenesis as well as androgen and estrogen signaling during in vitro maturation of mouse prepubertal testicular tissues. Histological, RT-qPCR, Western blot analyses, measurements of cholesterol, steroid hormones levels and aromatase activity were performed on fresh or frozen/thawed in vitro cultured mouse testicular tissues from 6.5 days postpartum (dpp) mice as well as on age-matched in vivo controls. A similar density of Leydig cells (LC) was found after 30 days of organotypic culture (D30) and at 36.5 dpp, the corresponding in vivo time point. However, LC were partially mature after in vitro culture, with decreased Sult1e1 and Insl3 mRNA levels (adult LC markers). Moreover, the transcript levels of Cyp11a1, Cyp17a1 and Hsd17b3 encoding steroidogenic enzymes were decreased in vitro. Increased amounts of progesterone and estradiol and reduced androstenedione intratesticular levels were observed at D30. Furthermore, androgen signaling was altered at D30, with decreased transcript levels of androgen target genes (Rhox5, Septin12). Moreover, the expression and activity of aromatase and estrogen signaling were impaired at D30. The addition of hCG to the organotypic culture medium induced an elevation in androgen production but did not improve sperm yield. In conclusion, this study reports partial LC maturation, disturbed steroidogenic activity of LC, abnormal steroid hormone content as well as altered androgen and estrogen signaling in cultures of fresh and frozen/thawed prepubertal mouse testicular tissues. The organotypic culture system will need to be further improved to increase the efficiency of in vitro spermatogenesis and allow a clinical application.

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“…Additionally, estrogen signaling was impaired in organotypic cultures, with a low expression of the estrogen-synthesizing enzyme aromatase, of estrogen receptors and of the estrogen target gene Faah ( Figure 7 ). Since the expression of Cyp19a1 , Esr1 and Esr2 (encoding aromatase, ERα and ERβ, respectively) can be downregulated by estradiol in the rat testis (61,62), we hypothesize that the transcription of these genes may be negatively controlled by elevated estradiol levels in our tissue cultures. Faah , whose promoter activity engages ERβ and the histone demethylase LSD1, is a direct target gene of estrogens in Sertoli cells (42).…”

Section: Discussionmentioning

confidence: 99%

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

Moutard

¹

,

Goudin

²

,

Jaeger

³

et al. 2022

Preprint

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0

View full text Add to dashboard Cite

Cancer treatments such as chemotherapy can have gonadotoxic effects. In order to preserve and restore the fertility of prepubertal patients with cancer, testicular biopsies are frozen and could theoretically be later matured in vitro to produce spermatozoa for assisted reproductive technology. A complete in vitro spermatogenesis has been obtained from prepubertal testicular tissue in the mouse model, although the sperm yield was low. Since steroid hormones play an essential role in spermatogenesis, it appears necessary to ensure that their synthesis and mechanisms of action are not altered in in vitro cultured tissues. The aim of this study was therefore to investigate steroidogenesis as well as androgen and estrogen signaling during in vitro maturation of mouse prepubertal testicular tissues. Histological, RT-qPCR, Western blot analyses, measurements of cholesterol, steroid hormones levels and aromatase activity were performed on fresh or frozen/thawed in vitro cultured mouse testicular tissues from 6.5 days postpartum (dpp) mice as well as on age-matched in vivo controls. A similar density of Leydig cells (LC) was found after 30 days of organotypic culture (D30) and at 36.5 dpp, the corresponding in vivo time point. However, LC were partially mature after in vitro culture, with decreased Sult1e1 and Insl3 mRNA levels (adult LC markers). Moreover, the transcript levels of Cyp11a1, Cyp17a1 and Hsd17b3 encoding steroidogenic enzymes were decreased in vitro. Increased amounts of progesterone and estradiol and reduced androstenedione intratesticular levels were observed at D30. Furthermore, androgen signaling was altered at D30, with decreased transcript levels of androgen target genes (Rhox5, Septin12). Moreover, the expression and activity of aromatase and estrogen signaling were impaired at D30. The addition of hCG to the organotypic culture medium induced an elevation in androgen production but did not improve sperm yield. In conclusion, this study reports partial LC maturation, disturbed steroidogenic activity of LC, abnormal steroid hormone content as well as altered androgen and estrogen signaling in cultures of fresh and frozen/thawed prepubertal mouse testicular tissues. The organotypic culture system will need to be further improved to increase the efficiency of in vitro spermatogenesis and allow a clinical application.

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“…The GO analysis based on AS data indicated that Sugp2 may participate in metal ion and energy metabolism. Combined with the results of KEGG, Sugp2 may be involved in calcium transport and signal transduction, which play an essential role in male germ cell development and function (34,35).…”

Section: Discussionmentioning

confidence: 99%

Chromatin-Associated Protein Sugp2 Involved in mRNA Alternative Splicing During Mouse Spermatogenesis

Zhan

¹

,

Li

²

,

Wu

³

et al. 2021

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Mammalian spermatogenesis is a highly ordered process that is determined by chromatin-associated moderators which still remain poorly understood. Through a multi-control group proteomics strategy, we confirmed that Sugp2 was a chromatin-associated candidate protein, and its signal arose along spermatogenesis. The expression results showed that Sugp2, which is mainly expressed in the testis, had two transcripts, encoding one protein. During spermatogenesis, Sugp2 was enriched in the nucleus of male germ cells. With the depletion of Sugp2 by CRISPER-Cas9 technology, we found that Sugp2 controlled a network of genes on metal ion and ATP binding, suggesting that alternative splicing regulation by Sugp2 is involved in cellular ion and energy metabolism during spermatogenesis, while it had a little effect on meiotic progression and male fertility. Collectively, these data demonstrated that, as a chromatin-associated protein, Sugp2 mediated the alternative splicing regulatory network during spermatogenesis.

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Estradiol and 1α,25(OH)2 vitamin D3 share plasma membrane downstream signal transduction through calcium influx and genomic activation in immature rat testis

Cited by 9 publications

References 51 publications

Steroidogenesis and androgen/estrogen signaling pathways are altered in in vitro matured testicular tissues of prepubertal mice

Steroidogenesis and androgen/estrogen signaling pathways are altered in in vitro matured testicular tissues of prepubertal mice

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

Chromatin-Associated Protein Sugp2 Involved in mRNA Alternative Splicing During Mouse Spermatogenesis

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