1-Nitropyrene exposure induces mitochondria dysfunction and impairs oocyte maturation in mice

Yu, Xiaoxia; Meng, Fei; Huang, Jianjun; Li, Weidong; Zhang, Jiaming; Yin, Shen; Zhang, Liangran; Wang, Shunxin

doi:10.1016/j.ecoenv.2022.113921

Cited by 10 publications

(2 citation statements)

References 80 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mitochondrial damage will activate the mitochondrial apoptosis program, which greatly limits the normal physiological function including protein synthesis. We herein used commercially available time-dependent mitochondrial damage agent 1-Nitropyrene (1-NP) and Carbonyl cyanide 3-chlorophenylhydrazone (CCCP) to induce mitochondrial apoptosis, atempting to explore the effect of mitochondria dysfuntion on protein expression. , To test whether mitochondrial dysfunction leads to decreased protein expression, we treated HEK293 cells with CCCP and 1-NP (1 μM) for 8, 24, and 48 h, respectively, before expressed SOD1(A4 V)-Halo and costaining with commercial mitochondrial labeling reagent (MitoTracker Green, 2 μM) and A9-Halo (2 μM). After treatment with MG132 (1 μM) for 24 h, a decreased fluorescent signal from A9-Halo was observed comparing to cells without treatment, suggesting there was a reduced level of protein expression (Figure a).…”

Section: Resultsmentioning

confidence: 99%

NRhFluors: Quantitative Revealing the Interaction between Protein Homeostasis and Mitochondria Dysfunction via Fluorescence Lifetime Imaging

Huang,

Chang,

Gao

et al. 2024

ACS Cent. Sci.

View full text Add to dashboard Cite

Degenerative diseases are closely related to the changes of protein conformation beyond the steady state. The development of feasible tools for quantitative detection of changes in the cellular environment is crucial for investigating the process of protein conformational variations. Here, we have developed a near-infrared AIE probe based on the rhodamine fluorophore, which exhibits dual responses of fluorescence intensity and lifetime to local viscosity changes. Notably, computational analysis reveals that NRhFluors fluorescence activation is due to inhibition of the RACI mechanism in viscous environment. In the chemical regulation of rhodamine fluorophores, we found that variations of electron density distribution can effectively regulate CI states and achieve fluorescence sensitivity of NRhFluors. In addition, combined with the AggTag method, the lifetime of probe A9-Halo exhibits a positive correlation with viscosity changes. This analytical capacity allows us to quantitatively monitor protein conformational changes using fluorescence lifetime imaging (FLIM) and demonstrate that mitochondrial dysfunction leads to reduced protein expression in HEK293 cells. In summary, this work developed a set of nearinfrared AIE probes activated by the RACI mechanism, which can quantitatively detect cell viscosity and protein aggregation formation, providing a versatile tool for exploring disease-related biological processes and therapeutic approaches.

show abstract

Section: Resultsmentioning

confidence: 99%

NRhFluors: Quantitative Revealing the Interaction between Protein Homeostasis and Mitochondria Dysfunction via Fluorescence Lifetime Imaging

Huang,

Chang,

Gao

et al. 2024

ACS Cent. Sci.

View full text Add to dashboard Cite

show abstract

“…1-NP is classified as a potential carcinogen by the International Agency for Research on Cancer (IARC), and the mutagenicity and carcinogenicity of 1-NP have been confirmed by many studies. − However, the mechanism of their transformation, the formation process of the main transformation products, and the contribution of hydroxylated products and epoxidation products to the health effects are less studied. Therefore, in this study, combining different isoenzymes with 1-NP, the main metabolic enzymes and the main risk conformations of 1-NP in vivo were analyzed.…”

Section: Introductionmentioning

confidence: 99%

1-Nitropyrene Metabolism and Health Risk: Identification of Key Enzymes, Pathways, and Metabolites Using a Multimethod Approach

Wang,

Gao,

Luo

et al. 2023

Environ. Health

View full text Add to dashboard Cite

Polycyclic aromatic hydrocarbon (PAH) derivatives have a widespread presence in the environment and even the human body, but their metabolism and potential risk remain unclear. In this study, we used molecular dynamics simulations and density functional theory to calculate the metabolic mechanism of 1-nitropyrene (1-NP), an important PAH derivative. The results showed that cytochrome P450 enzymes (CYPs) can metabolize 1-NP, with CYP 2A13 and CYP 2E1 being important enzyme isoforms, because they had lower binding affinities (−16.48 and −13.90 kcal/mol) to 1-NP than other CYPs (−2.38 to −7.89 kcal/mol). Additionally, these CYPs can metabolize 1-NP through epoxidation and hydroxylation pathways. Compared to hydroxylation, epoxidation had a lower energy barrier of 9.42 kcal/mol, and 4,5-epoxide-1-nitropyrene and 9,10-epoxide-1-nitropyrene were identified as the major epoxidation products through electrophilic addition. In addition, 6-hydroxy-1-nitropyrene and 8-hydroxy-1-nitropyrene were the major hydroxylated metabolites. Health risks revealed that electrophilicity of epoxides increased the risk of binding to DNA, and both 1-NP and its four important metabolites cause adverse effects on the gastrointestinal system and lung. In summary, this study revealed the metabolic mechanism of 1-NP by human CYPs and formation of toxic metabolites, and more attention should be paid to the nitro-derivatives of PAHs in the future.

show abstract

Employment of a Newly Defined In Vitro Fertilization Protocol to Determine the Cytoskeletal Machinery, DNA Damage, and Subsequent DNA Repair Resulting from Endocrine Disruption by Hexavalent Chromium in Rat Metaphase II Oocytes

Wuri,

Zarutskie,

Arosh

et al. 2024

Current Protocols

View full text Add to dashboard Cite

These protocols describe a detailed method to determine the DNA damage and F‐actin and microtubule defects of metaphase II oocytes caused by hexavalent chromium, Cr(VI), an endocrine disrupting chemical (EDC). The protocol provides systematic steps to determine protein expression encoded by pluripotency proteins such as Oct4, Nanog, and Cdx2 during early embryonic development. Occupational or environmental exposure to EDCs has significantly increased infertility in both men and women. The urinary concentration of the EDC bisphenol A in patients undergoing in vitro fertilization (IVF) is directly related to decreased implantation rates and the number of metaphase II oocytes recovered. This protocol outlines crucial steps in assessing the structure of F‐actin and microtubules, DNA damage, and repair mechanisms in metaphase II oocytes as well as pluripotency protein markers of early‐stage embryos. IVF techniques to achieve fertility goals in both humans and animals are of paramount importance. The interplay between F‐actin and microtubules is crucial for bipolar spindle assembly and correct partitioning of the nuclear genome in mammalian oocyte meiosis. EDCs induce DNA damage and impair DNA repair mechanisms, compromising oocyte quality. In human IVF, this results in failure to implant, early miscarriage, and live births with congenital disorders, thus decreasing success rates and increasing poor outcomes. The application of IVF protocols in rats to understand EDC‐mediated defects in the cytoskeletal network of metaphase II oocytes is not well established. We present a newly defined rat IVF protocol and demonstrate outcomes using these protocols to determine the adverse effects of Cr(VI) on metaphase II oocytes. Basic Protocol 1 includes steps to superovulate rats, dissect ampullae, retrieve oocytes/eggs, perform immunofluorescence staining of cytoskeletal machinery (microtubules and F‐actin), and assess expression of the DNA double‐strand break marker γ‐H2AX and the DNA repair protein RAD51 in control and Cr(VI)‐exposed rats. Basic Protocol 2 describes methods for detecting the pluripotency proteins Oct4, Nanog, and Cdx2 during early embryonic development in control rats. © 2024 Wiley Periodicals LLC.Basic Protocol 1: In vivo EDC treatment of rats and immunostaining of treated oocytesBasic Protocol 2: In vitro fertilization and immunostaining of early‐stage embryos

show abstract

1-Nitropyrene exposure induces mitochondria dysfunction and impairs oocyte maturation in mice

Cited by 10 publications

References 80 publications

NRhFluors: Quantitative Revealing the Interaction between Protein Homeostasis and Mitochondria Dysfunction via Fluorescence Lifetime Imaging

NRhFluors: Quantitative Revealing the Interaction between Protein Homeostasis and Mitochondria Dysfunction via Fluorescence Lifetime Imaging

1-Nitropyrene Metabolism and Health Risk: Identification of Key Enzymes, Pathways, and Metabolites Using a Multimethod Approach

Employment of a Newly Defined In Vitro Fertilization Protocol to Determine the Cytoskeletal Machinery, DNA Damage, and Subsequent DNA Repair Resulting from Endocrine Disruption by Hexavalent Chromium in Rat Metaphase II Oocytes

Contact Info

Product

Resources

About