Single-cell transcriptome dissection of the toxic impact of Di (2-ethylhexyl) phthalate on primordial follicle assembly

Abstract

Rationale: Accumulated evidence indicates that environmental plasticizers are a threat to human and animal fertility. Di (2-ethylhexyl) phthalate (DEHP), a plasticizer to which humans are exposed daily, can trigger reproductive toxicity by acting as an endocrine-disrupting chemical. In mammals, the female primordial follicle pool forms the lifetime available ovarian reserve, which does not undergo regeneration once it is established during the fetal and neonatal period. It is therefore critical to examine the toxicity of DEHP regarding the establishment of the ovarian reserve as it has not been well investigated. Methods: The ovarian cells of postnatal pups, following maternal DEHP exposure, were prepared for single cell-RNA sequencing, and the effects of DEHP on primordial follicle formation were revealed using gene differential expression analysis and single-cell developmental trajectory. In addition, further biochemical experiments, including immunohistochemical staining, apoptosis detection, and Western blotting, were performed to verify the dataset results. Results: Using single-cell RNA sequencing, we revealed the gene expression dynamics of female germ cells and granulosa cells following exposure to DEHP in mice. Regarding germ cells: DEHP impeded the progression of follicle assembly and interfered with their developmental status, while key genes such as Lhx8 , Figla, and others, strongly evidenced the reduction. As for granulosa cells: DEHP likely inhibited their proliferative activity, and activated the regulation of cell death. Furthermore, the interaction between ovarian cells mediated by transforming growth factor-beta signaling, was disrupted by DEHP exposure, since the expression of GDF9, BMPR1A, and SMAD3 was affected. In addition, DNA damage and apoptosis were elevated in germ cells and/or somatic cells. Conclusion: These findings offer substantial novel insights into the reproductive toxicity of DEHP exposure during murine germ cell cyst breakdown and primordial follicle formation. These results may enhance the understanding of DEHP exposure on reproductive health.

Protective Mechanism of Luteinizing Hormone and Follicle-Stimulating Hormone Against Nicotine-Induced Damage of Mouse Early Folliculogenesis

“…At this point, there are still plenty of oocytes remaining in the ovarian cysts. Consistent with previous studies Liu et al, 2019;Liu W. X. et al, 2021;Tian et al, 2021;Wang et al, 2021), the effect of the drug on germ cell cyst breakdown and PF formation could be measured by counting the proportion of oocytes in cysts and follicles. In addition, newborn mice were fed normally after 4 days of intraperitoneal injection.…”

“…In fact, many previous studies have confirmed that the obstruction of germ cell cyst breakdown and PF assembly are related to insufficient oocyte-specific gene expression (Wang Y. Y. et al, 2017;Wang et al, 2021;Wang J. J. et al, 2018;Liu J. C. et al, 2021). Interestingly, LH or FSH could reduce the effects of nicotine on specific gene expressions in oocytes, such as factor in the LIM homeobox 8 (Lhx8), growth differentiation factor-9 (Gdf9), spermatogenesis and oogenesis helix-loop-helix 2 (Sohlh2), and newborn ovary homeobox (Nobox; Figure 2A).…”

Exploring the Biological Activity and Mechanism of Xenoestrogens and Phytoestrogens in Cancers: Emerging Methods and Concepts

“…One group utilized scRNA-seq technology, although not directly related to cancer research, to investigate the transcriptomic changes induced by a known xenoestrogen, di (2-Ethylhexyl) phthalate (DEHP), exposure. They revealed the reproductive toxicity of DEHP in murine germ cells and pre-granulosa cells at a single-cell level [138]. Although scRNA-seq has some limitations, such as technical noise from the cell preparation process, loss of spatial information, higher costs than other models, and requirement for freshly prepared samples [139][140][141], it serves as an excellent option for studying the complicated activity of xenoestrogens/phytoestrogens in heterogeneous cell populations of target tissues.…”

“…Menopause is a critical WOS because of its hypersensitivity to endocrine-disrupting chemicals due to the decline of endogenous estrogen [208]. Studies from our group have discovered that PBDEs, the flame retardants in household products, enhance E2-mediated regrowth of mammary glands, augment E2-facilitated gene expression, and modulate immune regulation, thus increasing the risk of developing breast cancer [136][137][138][139]. Importantly, like the WOS in female breast cancer, there appears to be a heightened sensitivity of the prostate to these exogenous estrogens during the critical developmental windows, such as in utero, the neonatal period and puberty.…”

Combined Negative Effects of Microplastics and Plasticizer DEHP: the Increased Release of Nets Delays Wound Healing

“…Although a large number of MPs and DEHP are removed from the nal e uent of the sewage treatment plant, MPs (2.419×10 MP/d) and DEHP still enter the water environment every day, posing a threat to the health of sh and local residents (Takdastan, Niari et al 2021). Human and animal exposure to DEHP through ingestion, inhalation and skin absorption can cause liver and kidney damage (Camacho, Latendresse et al 2020), male and female reproductive disorders and developmental toxicity (Wang, Tian et al 2021). In addition, it is reported that DEHP induces the formation and apoptosis of carp neutrophil extracellular traps (Nets) by promoting ROS burst and autophagy (Yirong, Shengchen et al 2020).…”