Unveiling the Transgenerational Toxicity of TDtBPP in Zebrafish Using abinScience’s FOXO1 Protein

In the field of environmental toxicology, understanding the interactions between pollutants and biological macromolecules is critical for assessing ecological risks. A recent study by researchers from China University of Geosciences, published in Environmental Science & Technology on 2025-04-22, utilized abinScience’s Recombinant Human FOXO1 Protein (Catalog No. HC288012) to investigate the transgenerational toxicity of Tris(2,4-di-tert-butylphenyl) phosphate (TDtBPP), an emerging organic phosphate pollutant. TDtBPP, increasingly detected in aquatic environments due to its use as a flame retardant and plasticizer, poses potential risks to wildlife and human health. This study provides groundbreaking insights into how TDtBPP disrupts developmental pathways in zebrafish offspring, offering a scientific foundation for evaluating the broader ecological impacts of organic phosphates.

Fig.1.Schematic illustration showing TDtBPP exposure in female zebrafish, its impact on FOXO1/ripor2 signaling, and resulting malformation and death in offspring embryos

Experimental Design and Observed Developmental Impacts

The research team exposed 30-day-old female zebrafish to environmentally relevant concentrations of TDtBPP (0, 50, 500, and 5,000 ng/L) for 180 days, simulating chronic exposure scenarios in natural aquatic systems. Their findings revealed significant transgenerational effects on offspring embryos. In the high-dose group (5,000 ng/L), TDtBPP accumulation in embryos increased markedly, correlating with severe developmental abnormalities. Malformation rates rose by 2.5 times at 24 hours post-fertilization (hpf) and by 26.6 times at 72 hpf, while mortality rates surged by 3.6 times compared to the control group. These defects were first observed during the gastrula stage (8–9 hpf), a critical period for cell differentiation and tissue formation, manifesting as abnormalities in the enveloping layer—a protective cell layer essential for early embryogenesis. Consequently, key developmental metrics such as hatching rates and body length were significantly reduced, underscoring TDtBPP’s potent toxicity across generations.

Fig.2.zebrafish embryos at 24 hpf and 72 hpf, comparing normal, malformed, and dead embryos due to TDtBPP

Molecular Insights: The Role of FOXO1 and ripor2 in TDtBPP Toxicity

FOXO1, a forkhead box O transcription factor, is a key regulator of cellular processes such as apoptosis, oxidative stress response, and developmental gene expression. In zebrafish, FOXO1 plays a pivotal role in early embryogenesis by activating genes like ripor2, which is involved in cell migration, adhesion, and polarization—processes critical for proper tissue formation. Using abinScience’s high-purity Recombinant Human FOXO1 Protein, the researchers conducted transcriptomic analysis on TDtBPP-exposed embryos, revealing a significant downregulation of ripor2. To confirm this gene’s role, they performed ripor2 knockout experiments, which recapitulated the observed developmental defects, including abnormal enveloping layer formation and elevated embryo mortality. Further experiments demonstrated that TDtBPP directly binds to FOXO1, inhibiting its transcriptional activity and blocking the activation of ripor2. This disruption in the FOXO1/ripor2 signaling pathway provides a mechanistic explanation for the observed developmental toxicity, highlighting the sensitivity of early developmental stages to environmental pollutants.

Fig.3.Microscope images comparing control and shripor2 zebrafish embryos at 8 hpf and developmental stages from 9 hpf to 72 hpf, showing malformation due to ripor2 downregulation.

Binding Dynamics: TDtBPP’s Interaction with FOXO1

To elucidate the molecular interaction between TDtBPP and FOXO1, the study employed advanced techniques such as molecular docking and microscale thermophoresis (MST). Molecular docking revealed that TDtBPP binds to the same site on FOXO1 as the known inhibitor AS1842856, suggesting a competitive inhibition mechanism. MST experiments quantified this interaction, measuring a high binding affinity between TDtBPP and FOXO1, which confirmed that TDtBPP directly interferes with FOXO1’s ability to regulate downstream genes like ripor2. This competitive binding disrupts the normal transcriptional activity of FOXO1, leading to a cascade of developmental impairments in zebrafish offspring. These findings not only validate the utility of abinScience’s FOXO1 protein in such studies but also underscore the importance of understanding pollutant-protein interactions at a molecular level to predict and mitigate environmental risks.

Fig.4.Molecular docking images showing the binding of TDtBPP and AS1842856 to FOXO1, with interaction details including hydrogen bonds and alkyl interactions.

Advancing Environmental Toxicology Research with abinScience

abinScience’s Recombinant Human FOXO1 Protein (Catalog No. HC288012) played a crucial role in this study, enabling precise analysis of TDtBPP’s interaction with FOXO1 and its downstream effects. High-purity protein tools like this are essential for dissecting the molecular mechanisms of environmental pollutants, offering researchers the ability to:

• ✔Explore the molecular basis of pollutant-induced toxicity in model organisms like zebrafish, which share significant genetic homology with humans.
• ✔Support the development of predictive models for ecological risk assessment, aiding in the identification of high-risk pollutants before they cause widespread harm.
• ✔Facilitate translational research by providing insights into potential human health risks, such as developmental disorders linked to environmental exposures.
• ✔Enable high-throughput screening of other organic phosphates to identify additional pollutants with similar toxicity profiles, informing proactive environmental management strategies.

The findings from this study pave the way for future research into the transgenerational effects of environmental pollutants, particularly in the context of developmental toxicology. For a deeper dive into the study, the original publication summary is available here. abinScience remains committed to supporting such groundbreaking research with reliable, high-quality tools tailored for environmental toxicology studies.^[web:0]

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