Identification of internal reference genes for porcine immature Sertoli cells under heat stress

Wang, Yi; Wu, Zi‐Wei; Fang, Ting; Zhang, Yuqing; Chen, Lu; Du, Zhiqiang; Yang, Caixia

doi:10.1111/rda.14211

Cited by 5 publications

(2 citation statements)

References 39 publications

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“…Our results that the expression levels of functional factors of Sertoli cell origin such as ABP, inhibin, AR, and GDNF that are involved in the development of spermatogenesis were significantly higher at 35 • C than at 37 • C. These results support the findings of previous studies showing that an increase in temperature affected Sertoli cell functionality, producing less ABP [25,40,41]. Furthermore, acute heat stress (37 • C, 0.5 h) decreased the viability of porcine immature Sertoli cells in vitro [42]. In addition, a single heat shock leads to changes in testicular weight and gene expression [43].…”

Section: Discussionsupporting

confidence: 88%

Effect of Temperature on the Development of Stages of Spermatogenesis and the Functionality of Sertoli Cells In Vitro

Jorban,

Lunenfeld,

Huleihel

2024

IJMS

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Spermatogenesis is the process of proliferation and differentiation of spermatogonial cells to meiotic and post-meiotic stages and sperm generation. Normal spermatogenesis occurs in vivo at 34 °C to 35 °C, and high temperatures are known to cause male infertility. The aim of the present study was to examine the effect of temperature (35 °C compared to 37 °C) on the viability/apoptosis of developed cells, on the development of different stages of spermatogenesis in 3D in vitro culture conditions, and the functionality of Sertoli cells under these conditions. We used isolated cells from seminiferous tubules of sexually immature mice. The cells were cultured in methylcellulose (as a three-dimensional (3D) in vitro culture system) and incubated in a CO2 incubator at 35 °C or 37 °C. After two to six weeks, the developed cells and organoids were collected and examined for cell viability and apoptosis markers. The development of different stages of spermatogenesis was evaluated by immunofluorescence staining or qPCR analysis using specific antibodies or primers, respectively, for cells at each stage. Factors that indicate the functionality of Sertoli cells were assessed by qPCR analysis. The developed organoids were examined by a confocal microscope. Our results show that the percentages and/or the expression levels of the developed pre-meiotic, meiotic, and post-meiotic cells were significantly higher at 35 °C compared to those at 37 °C, including the expression levels of the androgen receptor, the FSH receptor, transferrin, the androgen-binding protein (ABP), and the glial-derived nerve growth factor (GDNF) which were similarly significantly higher at 35 °C than at 37 °C. The percentages of apoptotic cells (according to acridine orange staining) and the expression levels of BAX, FAS, and CASPAS 3 were significantly higher in cultures incubated at 37 °C compared to those incubated at 35 °C. These findings support the in vivo results regarding the negative effect of high temperatures on the process of spermatogenesis and suggest a possible effect of high temperatures on the viability/apoptosis of spermatogenic cells. In addition, increasing the temperature in vitro also impaired the functionality of Sertoli cells. These findings may deepen our understanding of the mechanisms behind optimal conditions for normal spermatogenesis in vivo and in vitro.

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Section: Discussionsupporting

confidence: 88%

Effect of Temperature on the Development of Stages of Spermatogenesis and the Functionality of Sertoli Cells In Vitro

Jorban,

Lunenfeld,

Huleihel

2024

IJMS

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“…The testicles of mice were bathed in water at 40 • C for 20 min, which resulted in a decrease 2 of 17 in germ cell proliferation and a decrease in circulating testosterone [14]. Acute heat stress (43 • C, 0.5 h) can reduce the cell viability of porcine immature Sertoli cells (iSCs) cultured in vitro [15]. The changes in gene expression can be detected in mouse testes at 4 h after a single heat shock at 43 • C for 20 min and the weight of testes decreases significantly, which still cannot be recovered on the 68th day [16].…”

Section: Introductionmentioning

confidence: 99%

Potential Function of Testicular MicroRNAs in Heat-Stress-Induced Spermatogenesis Disorders

Gan

Xie

et al. 2023

IJMS

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Spermatogenesis is temperature-dependent, and the increase in testicular temperature seriously affects mammalian spermatogenesis and semen quality. In this study, the testicular heat stress model of mice was made with a 43 °C water bath for 25 min, and the effects of heat stress on semen quality and spermatogenesis-related regulators were analyzed. On the 7th day after heat stress, testis weight shrank to 68.45% and sperm density dropped to 33.20%. High-throughput sequencing analysis showed that 98 microRNAs (miRNAs) and 369 mRNAs were down-regulated, while 77 miRNAs and 1424 mRNAs were up-regulated after heat stress. Through gene ontology (GO) analysis of differentially expressed genes and miRNA–mRNA co-expression networks, it was found that heat stress may be involved in the regulation of testicular atrophy and spermatogenesis disorders by affecting cell meiosis process and cell cycle. In addition, through functional enrichment analysis, co-expression regulatory network, correlation analysis and in vitro experiment, it was found that miR-143-3p may be a representative potential key regulatory factor affecting spermatogenesis under heat stress. In summary, our results enrich the understanding of miRNAs in testicular heat stress and provide a reference for the prevention and treatment of heat-stress-induced spermatogenesis disorders.

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Selection of reference genes for RT-qPCR analysis in developing chicken embryonic ovary

Wang

Zhang

et al. 2023

Mol Biol Rep

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Identification of internal reference genes for porcine immature Sertoli cells under heat stress

Cited by 5 publications

References 39 publications

Effect of Temperature on the Development of Stages of Spermatogenesis and the Functionality of Sertoli Cells In Vitro

Effect of Temperature on the Development of Stages of Spermatogenesis and the Functionality of Sertoli Cells In Vitro

Potential Function of Testicular MicroRNAs in Heat-Stress-Induced Spermatogenesis Disorders

Selection of reference genes for RT-qPCR analysis in developing chicken embryonic ovary

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