Perfluorohexane sulfonate (PFHxS), an increasingly detected environmental pollutant, poses potential metabolic health risks, yet its developmental toxicity at environmentally relevant doses remains poorly understood. Here, we investigated the long-term effects of PFHxS exposure on the hepatic development using a mouse model with oral administration of 0.03 and 0.3 μg/kg/day throughout pregnancy. Our findings demonstrated that gestational PFHxS exposure induced persistent hepatic oxidative stress and was associated with the activation of the endoplasmic reticulum stress response (PERK/eIF2α/ATF4 pathway) in adults. These molecular alterations were accompanied by the upregulation of sterol regulatory element-binding protein 1 (SREBP1) and the transcriptional activation of key lipogenic enzymes (ACC, FASN, SCD1), culminating in hepatic lipid accumulation, inflammation, and fibrosis. Notably, these pathological changes exhibited sex-specific characteristics. These findings revealed, for the first time, that gestational PFHxS exposure at environmental levels may contribute to adult hepatic steatosis, potentially through mechanisms involving PERK/SREBP1-mediated lipogenesis. This study provided critical insights into the emerging health risks of replacement PFAS compounds.

The role and underlying mechanisms of placental ferroptosis in fetal growth restriction (FGR) induced by environmental stress remain poorly understood. Our population-based study showed elevated ferroptosis levels in all-cause FGR placentae. Environmental stressor cadmium (Cd) was used to generate an FGR mouse model, which exhibited elevated placental ferroptosis. Ferroptosis inhibitor ferrostatin-1 reversed environmental Cd-induced FGR. Targeted oxidized lipidomics identified the peroxisome as a target organelle for prenatal Cd-induced placental ferroptosis. Furthermore, Cd induced excessive activation of PEX5-dependent pexophagy in placentae. By establishing a placental Pex5-knockdown mouse, pexophagy was confirmed to drive environmental Cd-induced placental ferroptosis. Mechanistically, pexophagy drives the degradation of the H2O2-scavenging enzyme and the fatty-acid β-oxidation enzymes, thereby causing lipid peroxidation and placental ferroptosis. Notably, environmental Cd upregulated PEX2, an E3 ligase mediating PEX5 monoubiquitination, thereby driving pexophagy and placental ferroptosis. Furthermore, METTL14-mediated m6A modification enhanced the stability of placental PEX2 mRNA in an ELAVL1-dependent manner under environmental Cd. SAH, a METTL14 inhibitor, alleviated Cd-induced placental pexophagy, ferroptosis, and FGR. High-temperature also decreased GPX4 and increased PEX2, PEX5, and METTL14 in placentae. Overall, our findings uncover a novel m6A-PEX2-PEX5 axis driving pexophagy-dependent placental ferroptosis, offering placental pexophagy as a therapeutic target for FGR and fetal-origin adult diseases.

Perfluorohexane sulfonate (PFHxS), a persistent organic pollutant and developmental toxicant, poses significant health concerns. However, the long-term impacts of human-relevant prenatal PFHxS exposure on female reproductive development remain unclear. This study investigated its effects on ovarian development and estrous cyclicity outcomes in female adolescent using a mouse model. Pregnant mice were exposed to human relevant doses of PFHxS via oral gavage throughout gestation. Our findings showed that prenatal PFHxS exposure significantly delayed vaginal opening (VO) and disrupted estrous cyclicity in the adolescent, with prolonged diestrus. Ovarian histology revealed impaired folliculogenesis, including increased atretic follicles and reduced corpus lutea. Serum hormone analyses showed disrupted steroidogenesis with elevated testosterone and estradiol (E2), yet decreased anti-Müllerian hormone (AMH) and follicle-stimulating hormone (FSH). Ovarian transcriptomics identified dysregulation in steroid biosynthesis pathways (e.g., Cyp11a1, Cyp19a1, and Amh), validated by qRT-PCR. In silico simulations demonstrated PFHxS-induced structural reconfiguration of CYP11A1 and CYP19A1 substrate-binding sites. These findings demonstrated that prenatal PFHxS exposure altered female reproductive development by disrupting ovarian steroidogenesis and folliculogenesis. Our study provided experimental evidence for the risk assessment of PFHxS and underscores the potential health implications of this persistent environmental contaminant.

Chronic kidney disease (CKD) represents one of the global public health challenges. Several epidemiological reports indicate that environmental arsenic exposure is associated with CKD. This study aimed to investigate the effect of long-term arsenic exposure on renal lipid metabolism during the development of kidney fibrosis. Adult C57BL/6 mice were administered NaAsO2 (3 or 15 mg/L) in drinking water for 6 months. Urinary protein, NGAL, α1-MG and β2-MG, markers of tubular injury, were increased in arsenic-exposed mice. In the later stage, renal inflammation and epithelial-mesenchymal transition indicators remained elevated. Collagen genes, including Col1a1, Col1a2, Col11a1, and Col11a2, were upregulated and collagen deposition was aggravated in the kidneys of arsenic-exposed mice. Mechanistically, renal lipid composition, as assessed by lipidomics, was significantly altered in arsenic-exposed mice. In the later stage, renal free fatty acids (FFAs) remained increased. Renal triglyceride (TG) content was elevated, and lipid droplet deposition was observed. Transcriptome sequencing showed that fatty acid oxidation (FAO) pathways were enriched in arsenic-exposed mouse kidneys. Real-time RT-PCR found that renal FAO-related genes were downregulated in arsenic-exposed mice. These results indicate that long-term arsenic exposure induces kidney fibrosis accompanied by renal lipid droplet deposition. Renal lipid metabolic disorders might be a mediating mechanism underlying arsenic-induced kidney fibrosis.

Male infertility affects approximately one in seven couples worldwide. Prenatal cadmium (Cd) exposure has been shown to affect offspring phenotypes and increase susceptibility to diseases later in life. However, the effects of prenatal Cd exposure on multi-generational offspring fertility and the mechanisms remain unknown. A novel murine multi-generational (F1-F3 offspring) male subfertility model induced by prenatal Cd exposure was developed. The levels of testosterone and steroidogenic enzymes were also lower in these offspring's testes. The ubiquitin-dependent degradation of NR4A1, the upstream transcription factor regulating steroidogenic enzymes, was enhanced across generations upon prenatal Cd exposure. After treatment with MG132, an inhibitor of the ubiquitin-proteasome system, the levels of NR4A1 and steroidogenic enzymes were higher in offspring testes with prenatal Cd exposure. Based on the analysis of the UbiBrowser database and testicular global transcriptome, RAPSN was identified as a novel ubiquitin E3 ligase containing the RING-H2_Rapsyn domain that mediates multi-generational testicular NR4A1 ubiquitination. m6A epitranscriptome analysis revealed that prenatal Cd exposure upregulated RAPSN expression in multi-generational offspring testes, and was attributed to a higher level of m6A modification of Rapsn mRNA. Furthermore, there was a lower level of YTHDC2, a m6A reader, in the multi-generational offspring testes with prenatal Cd exposure. Prenatal and postnatal testicular YTHDC2 overexpression reduced the stability of m6A-methylated Rapsn mRNA to downregulate RAPSN expression in F1-F3 testes. Overall, YTHDC2 reduction-mediated increment in m6A-methylated Rapsn mRNA contributed to prenatal Cd-enhanced multi-generational susceptibility to male subfertility.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social impairments and stereotyped behaviors. Many epidemiological studies have found a potential relationship between vitamin D deficiency (VDD) and ASD. However, the mechanism remains unclear. In this study, a VDD model was established by feeding a vitamin D-depleted diet to 5-week-old female mice, from their adulthood through pregnancy to end of lactation. Social deficits, repetitive stereotyped behaviors and anxiety-like behaviors were evaluated in the offspring. The results showed the number of buried marbles was increased in the female offspring of the VDD group. Social defects were observed in both male and female offspring in the VDD group. Mechanistically, several markers of cell proliferation, such as Pcna and Ki67, were upregulated. And the number of TBR2+ cell, an intermediate progenitor cell, was increased in cerebral cortex of VDD-fed fetuses. Moreover, DKK1, a WNT/β-catenin pathway repressor, was elevated in cerebral cortex of VDD-fed fetuses. By contrast, β-catenin, a critical effector of the WNT/β-catenin pathway, was reduced in cerebral cortex of GD14 VDD fetuses. These results provide partial evidence that maternal vitamin D deficiency during pregnancy and lactation induces autism-like behaviors partly by suppressing WNT/β-catenin pathway in the cerebral cortex.

Environmental stressors-induced male infertility has become a major public health issue. Sperm motility is the key to conception. However, the immunological mechanism for environmental stress-induced sperm motility reduction remains unknown. Our experiments find that three classical environmental stressors, including lead, cadmium, and mercury, are the key heavy metals reducing sperm motility. We reveal that environmental stress induces epididymal macrophage senescence, whereas the clearance of above cells significantly alleviates environmental stress-reduced sperm motility. In vivo and in vitro experiments further demonstrate that environmental stress induces macrophage senescence via down-regulating Foxo3 expression. Mechanistically, environmental stress increases the epididymal Foxo3 N6-methyladenosine level and inhibits the FTO. Overexpression of Fto in vitro and S-adenosylhomocysteine supplementation in vivo alleviates environmental stress-induced macrophage senescence. Together, these results suggest that epididymal macrophage senescence contributes to sperm motility decrease upon environmental stress. Clearing senescent epididymal macrophages will provide a previously unknown strategy for preventing male infertility.

The plasticizer DEHP is ubiquitous in the environment, and its unavoidable daily low-dose chronic exposure can impair male sperm. Paternal sperm damage is associated with abnormal placental development in offspring. However, the underlying mechanisms remain unknown. This study innovatively explored the effects and potential mechanisms of paternal DEHP exposure on placenta development in offspring. The results showed that 90-day exposure to environmental doses of DEHP, male mice exhibited sperm damage, accompanied by a significant reduction in the vascular sinus area within the labyrinthine layer of GD17 placentas, indicating that DEHP-induced sperm damage impairs placental angiogenesis in offspring. Transcriptomic revealed the mechanism of placental angiogenesis obstruction, we found that in both fertilized zygotes and GD17 placentas, the mRNA and protein expression of the angiogenesis-associated type I collagen genes Col1α1 were significantly decreased. Furthermore, the expression of binding partner integrin α2β1 was decreased, leading to inhibition of downstream PI3K-AKT signaling pathway activity. To verify that reduced expression of Col1α1 and Col1α2 inhibits placental angiogenesis, the study knocked down Col1α1 and Col1α2 in Human Umbilical Vein Endothelial Cells (HUVEC), and confirmed the inhibitory effect on angiogenesis and related signaling pathways. In summary, this study demonstrates that paternal DEHP exposure may induce downregulation of Col1α1 and Col1α2, which are important angiogenic genes in the fertilized zygotes and placenta, leading to a reduction in placental sinus area. This study clarifies the association between paternal subchronic exposure to environmental doses of DEHP and abnormal placental vascular development in offspring, challenging the traditional “maternal dominance” paradigm in placental development research. The study provides an important scientific basis for comprehensively assessing the reproductive health risks of environmental pollutants and for formulating precise prevention and control strategies.

Air pollution control has lowered major criteria pollutants, yet airborne organic pollutants remain understudied, particularly regarding biomarkers and risks in pregnancy. We characterized 20 urinary biomarkers of airborne organic pollutants, including p-phenylenediamines, nitrated polycyclic aromatic hydrocarbons, phenylguanidines, benzothiazoles/benzotriazoles, and cotinine, in pregnant women from the prospective Towards Improved Maternal and Fetal health via Multipoint Exposure Monitoring (TIMFEM) study in China. Morning urine samples were analyzed by LC-MS/MS. Epidemiologic associations were integrated with in vitro screening in human trophoblast cells using a leave-one-out approach at human-exposure-guided doses. Among 1425 mother-infant pairs, 18 biomarkers were detected in >40% of participants. Each natural log-unit increase in N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD) and its quinone (6PPD-Q) was associated with 27% and 23% higher risk of small-for-gestational-age (SGA), respectively. Mixture models identified 1,2,3-triphenylguanidine (TPG), cotinine, and 1-aminopyrene as additional key contributors. Although seven priority pollutants showed minimal effects on cell proliferation, they reduced pyruvate entry into the tricarboxylic acid cycle and limited isocitrate-to-α-ketoglutarate conversion, indicating impaired placental energy metabolism. Overall risk ranking highlighted cotinine, TPG, and 6PPD-Q. Maternal coexposure to airborne organic pollutants was associated with elevated SGA risk, with metabolic disruption, rather than acute cytotoxicity, suggesting a plausible mechanistic pathway.

BACKGROUND:Early-life exposure to exogenous chemicals can disrupt neurodevelopment. Perfluorohexanesulfonic acid (PFHxS), a legacy PFAS widely used and detected globally, remains poorly studied for its neurotoxicity. METHODS:CD-1 mice (n = 90 dams) were exposed to human-relevant dose of PFHxS from gestational day (GD) 0-17. PFHxS levels were measured in maternal plasma and foetal/offspring medial prefrontal cortex (mPFC) (GD 18, postnatal weeks [PNW] 4 and 10) using liquid chromatography tandem mass-spectrometry (LC-MS/MS). Offspring social behaviour was assessed with the Three-Chamber social test. Neurotransmitters in mPFC of PNW 10 offspring were profiled by LC-MS/MS, transcriptomics was performed on GD 18 and PNW 4 mPFC, GABAergic neurons were quantified by immunofluorescence, and glutamate decarboxylase (GAD) expression by western blotting. PFHxS-GAD interactions were examined via molecular docking and microscale thermophoresis (MST). FINDINGS:Maternal plasma reached 5.1 ± 0.1 ng/mL, equivalent to human biomonitoring data, and PFHxS accumulated in foetal mPFC (68.1 ± 4.1 pg/g). PFHxS exposure induced social deficits at PNW 4 and 10, which were more pronounced in males. The mPFC of PFHxS exposed offspring exhibited an excitatory-tilted neurotransmitter profile, feature with reduced γ-aminobutyric acid (GABA, 90.6 ± 12.2 vs. 70.2 ± 4.3 μg/g, p = 0.008). The differentially expressed genes were enriched in GABAergic and synaptic signalling pathways. Despite unchanged percentage of GABAergic neuron and GAD expression, metabolite GABA/glutamate ratio was attenuated, suggesting impaired GAD function. Both molecular docking and MST suggested moderate-to-strong binding affinity between PFHxS and GAD, affecting GABA synthesis. INTERPRETATION:Gestational PFHxS exposure at human-relevant levels impairs offspring social behaviour, likely via disruption of GAD-mediated imbalance in excitation/inhibition. FUNDING:This work was provided by grants from the National Natural Science Foundation of China (82404221 and 82373586), Anhui Provincial Natural Science Foundation (2308085Y50 and 2408085QH275), Education Department of Anhui Province for Excellent Young Scientist (2022AH030076), and funding from Center for Big Data and Population Health of IHM (JKS2022020).

The widespread detection of PFOS precursors in the environment and human matrices raises a new wave of health concerns. However, their absorption, distribution, metabolism, and excretion (ADME) fates and toxicity profiles in mammals remain largely unexplored. Here, we investigated the toxicokinetic, metabolic transformation, and toxicity of three precursors, N-MeFOSA, N-EtFOSA, and N-EtFOSAA, by integrating toxicokinetic modeling in mice, rat liver microsome metabolism, and hepatocellular toxicity assays. N-MeFOSA and N-EtFOSA were rapidly eliminated in vivo and exhibited high efficiencies of conversion to legacy PFAS in liver microsomes, surpassing the typically low transformation rates in mammals reported for other precursors. Notably, N-MeFOSA showed the greatest metabolic transformation potential (25% legacy PFAS and 22% PFOS in 3 h). Moreover, in hepatocellular toxicity assays, both precursors exhibited weaker direct metabolic perturbations than their metabolites (PFOS and FOSA), indicating that their toxicity is primarily enhanced via metabolic activation. In contrast, N-EtFOSAA displayed persistence and stability in vivo and in vitro and induced lipid accumulation comparable to that of PFOS, together with stronger inhibition of cellular energy metabolism. Collectively, our study provides crucial ADME data identifying N-alkyl-FOSA-type precursors as non-negligible internal PFOS sources, underscoring a dual-risk paradigm where toxicity is driven either by metabolic activation or intrinsic parent compound toxicity.

Gestational exposure to cadmium (Cd), a widespread environmental toxicant, disrupted placental angiogenesis to induce fetal growth restriction (FGR). This study aimed to clarify the mechanism by which Cd disrupts placental angiogenesis. Human and mouse studies indicated that Cd exposure reduced VEGF-A to disrupt placental angiogenesis. Further data confirmed that gestational Cd exposure promoted estrogen receptor ESR1-specific degradation via ubiquitin-proteasome system (UPS), thereby decreasing placental VEGF-A. Bortezomib, the only clinically approved protease inhibitor, blocked ESR1 degradation to alleviate Cd-impaired placental angiogenesis. Based on mouse and human transcriptomics, WWP2 was identified as an unreported ubiquitin E3 ligase targeting placental ESR1. Specifically, both WWP2 knockdown and its inhibitor NSC2805 treatment consistently reversed environmental Cd-induced placental angiogenesis disorders and FGR. Furthermore, the m6A modification in Wwp2 mRNA was increased in Cd-exposed placentae. METTL3 and ELAVL1 knockdown verified that m6A modification enhanced the stability of Wwp2 mRNA. SAH, an inhibitor for METTL3, not only decreased WWP2 but also alleviated Cd-impaired placental angiogenesis and fetal growth. Based on a human case-control study, m6A-methylated Wwp2 was positively correlated with placental angiogenesis inhibition and all-cause FGR. In conclusion, gestational Cd exposure enhanced m6A modification in Wwp2 mRNA to drive ESR1 degradation, thereby inhibiting placental angiogenesis and fetal growth.

Humans are increasingly exposed to "eco-friendly" biodegradable microplastic pollution, whose usage in packaging and medical applications is growing exponentially. The bioplastic polylactic acid (PLA) has recently been demonstrated to release large quantities of oligomeric lactic acid (OLA) nanoplastics causing adverse health effects. No research has reported on intrauterine biodistribution of OLA, and how gestational exposure may impact on early development of the fetus. Here, we reveal that OLA plastics can readily breach the placental barrier and accumulate in various fetal organs in a mouse model. Gestational exposure to environmentally relevant dose of OLA impairs vasculature development, causing intrauterine growth restriction in the pups. Mechanistically, OLA causes blockage of the vascular endothelial growth factor pathway and abnormal physiological development of placenta, which is mediated by the obstruction of transcription factor GATA2 translocation into the nucleus. This study highlights the potential developmental health effect of oligomer nanoparticles released from biodegradable PLA plastic.

Cadmium (Cd), a bioaccumulative toxic heavy metal, poses a growing concern for offspring reproductive health following gestational exposure. Premature ovarian insufficiency (POI), a major cause of female infertility, remained poorly understood in terms of environmental etiology. This study investigated whether gestational Cd exposure induced ovarian functional decline via epigenetic mechanisms in mice. Using a mouse model of maternal Cd exposure via drinking water, we found that gestational Cd exposure led to POI-like ovarian dysfunction in adult offspring, characterized by diminished ovarian reserve, disrupted estrous cycles, altered serum hormone profiles, and accelerated ovarian aging with fibrosis and DNA damage. Mechanistically, Cd exposure upregulated DNMT1 and DNMT3A expression in offspring ovaries, promoting hypermethylation of the Klotho (Kl) promoter and suppressing its expression. Kl downregulation subsequently attenuated its inhibition of the p53/p21 pathway, triggering granulosa cell senescence and premature follicular depletion. Importantly, administration of the DNA methylation inhibitor 5-aza-2′-deoxycytidine (5-Aza) during gestation partially reversed these Cd-induced epigenetic modifications and ameliorated the ovarian phenotypes in offspring. Our findings revealed a complete pathway through which gestational Cd exposure promoted ovarian aging via DNMT-mediated Kl silencing and p53/p21 activation. This work provided novel mechanistic insights into environmental reproductive toxicity and identified potential targets for intervention. Environmental implication: Cd is a pervasive environmental toxicant threatening female reproductive health, yet its long-term developmental impacts on offspring ovarian aging remain unclear. This study reveals that gestational Cd exposure epigenetically silences the anti-aging gene Kl via promoter hypermethylation, activating the p53/p21/p16-Rb senescence pathway and triggering POI-like ovarian dysfunction in offspring. Importantly, this epigenetic damage is partially reversible with 5-Aza, offering a potential intervention strategy. These findings provide mechanistic insights into how Cd pollution contributes to rising POI incidence, informing developmental origins of reproductive dysfunction and guiding preventive policies for hazardous material regulation.

The role of placental cellular senescence in environmental cadmium (Cd)-evoked fetal growth restriction (FGR) and its underlying mechanisms require further clarification. Here, we generated a murine FGR model by simulating internal exposure doses of Cd in humans. Human and mouse studies revealed that placental senescence linked environmental Cd exposure to FGR. Furthermore, environmental Cd degraded mitochondrial anti-aging protein SIRT3 to evoke placental cellular senescence and FGR, as demonstrated by SIRT3 overexpression and its activator resveratrol treatment. Interestingly, CLPP was identified as a mitochondrial protease targeting placental SIRT3 degradation under environmental Cd. In vitro CLPP knockdown and in vivo CLPP inhibitor tamarixetin treatment reversed environmental Cd-induced SIRT3 degradation and placental cellular senescence. Additionally, environmental Cd elevated the level of METTL3 protein to promote m6A modification of ClpP mRNA in placentae. In vitro METTL3 knockdown and in vivo its inhibitor S-Adenosylhomocysteine treatment blocked the activation of CLPP-dependent mitochondrial protease stress, attenuating placental cellular senescence and FGR upon environmental Cd. Based on a human case-control study, m6A-driven CLPP-dependent mitochondrial protease stress was positively correlated with placental cellular senescence and all-cause FGR. Taken together, environmental Cd enhances m6A modification to activate CLPP-dependent mitochondrial protease stress, thereby causing placental cellular senescence and FGR.

Fenvalerate, a representative type II pyrethroid insecticide, is well established in the literature. Fenvalerate exerts developmental and neurological toxicity. We assessed whether paternal fenvalerate exposure induces autism-like behavioral alterations in offspring using a mouse model. Behavioral tests, including the three-chamber social interaction, self-grooming, and marble-burying tests, showed altered social and repetitive behaviors in offspring from fenvalerate-exposed fathers. NeuN, a mature neuronal marker, was reduced in the medial prefrontal cortex (mPFC) of weaning offspring from paternal fenvalerate-exposed groups. Nissl staining showed that the number of surviving neurons is reduced in the mPFC of weaning pups with paternal exposure to fenvalerate. Nestin, a marker for neural stem cells, was decreased in the fetal forebrain from paternal fenvalerate-exposed groups. Transcriptome analysis and RT-PCR showed that insulin-like growth factor 2 (IGF2), a neurotrophic factor, was downregulated in the paternal fenvalerate-exposed group. IGF2 protein was reduced in the fetal forebrain from paternal fenvalerate-exposed group. In addition, paternal fenvalerate exposure reduced methylation of the IGF2 imprinted control region (ICR) in fetal forebrain and paternal sperm. In conclusion, autism-like behaviors appear in offspring after paternal exposure to fenvalerate, which may partly be related to disruptions in epigenetic reprogramming of IGF2 in the developing brain and paternal sperm.

Metabolic dysfunction-associated fatty liver disease (MAFLD) refers to hepatic steatosis accompanied by one of the following: type 2 diabetes, obesity, overweight, or metabolic dysfunction. Developmental arsenic (As) exposure is linked to metabolic disorders, but its effects on MAFLD-related phenotypes remain unclear. This study aims to assess the impact of prenatal As exposure on MAFLD-like phenotypes in adulthood offspring. Pregnant mice were given deionized water containing NaAsO2 at concentrations of 0, 1.5, and 15 mg/L throughout their pregnancy. Doses were selected based on environmentally relevant exposure (1.5 mg/L, L-As) and prior evidence of metabolic effects in rodents (15 mg/L, H-As). Gestational H-As exposure increased hepatic triglyceride content and enlarged hepatic lipid droplets in middle-aged offspring. Intraperitoneal glucose tolerance test and insulin tolerance test analyses revealed that prenatal L-As and H-As exposure impaired glucose and insulin tolerance in middle-aged offspring. White fat mass and adipocyte size were elevated in H-As exposed middle-aged offspring. Several metabolism-related hormones, including adiponectin, leptin, and insulin, were elevated, whereas glucagon-like peptide-1, a hormone secreted by the intestines, was reduced in H-As exposed female middle-aged offspring. These results demonstrate that prenatal As-alone promotes MAFLD-like phenotypes in middle-aged offspring, highlighting its role as an early life environmental trigger for metabolic disease.

Recently, the perspective of paternal origin has emerged, yet its role in motor disorders remains unclear. Here, using Drosophila and murine models, we demonstrated that following paternal environmental heavy metal stress, offspring from multiple generations exhibited progressive motor deficits accompanied by dopaminergic neuron loss. Cross-tissue RNA sequencing revealed dysregulation of CG9593 (functionally homologous to human angiopoietin-like protein 4, ANGPTL4), identifying CG9593/ANGPTL4 as a pivotal brain effector linking paternal heavy metal stress to multigenerational motor disorders across species. Specifically, neuron-specific CG9593 knockdown ameliorated these disorders, whereas CG9593 overexpression mimicked them. Mechanistically, N6-methyladenosine (m6A) hypermethylation in sperm stabilized CG9593 messenger RNA in offspring neurons via IGF2BP1-dependent posttranscriptional regulation, reducing synapse-associated protein DLG1 and impairing motor neuron. Human cohort analyses confirmed that serum ANGPTL4 negatively correlated with pediatric motor scores, and paternal sperm m6A levels were positively associated with the heavy metal exposure burden. Our findings establish CG9593/ANGPTL4 as an evolutionarily conserved determinant of paternally acquired multigenerational motor disorders and hold promise as a druggable target.

Arsenic (As) is an environmental metalloid. Previous studies have demonstrated that As exposure resulted in decline of sperm quality. This study aimed to investigate the impact of exposure to As on blood-testis barrier (BTB) in a mouse model. Four-week-old male mice were exposed to NaAsO2 (1 or 15 mg/L) for 6 weeks. Our results found that NaAsO2 exposure disrupted the BTB and reduced sperm counts in adult mice. NaAsO2 activated the integrated stress response (ISR) and downregulated barrier junction protein in mouse testes and Sertoli cells. Ribosome profiling sequencing (Ribo-seq) and Ribosome-nascent chain complex-bound mRNA qPCR (RNC-qPCR) showed that translational efficiency of N-cadherin and ZO-1, two key barrier junction proteins, was reduced in NaAsO2-treated Sertoli cells. Mechanistically, NaAsO2 exposure reduced SIRT3 protein via proteasomal degradation, thereby resulting in mitochondrial dysfunction and excess mitochondrial ROS (mtROS) generation in Sertoli cells. Melatonin alleviated NaAsO2-induced mitochondrial dysfunction and mtROS upregulation via reducing SOD2 acetylation in Sertoli cells. Moreover, melatonin antagonized NaAsO2-induced ISR, barrier junction proteins downregulation and barrier function impairment in Sertoli cells. Accordingly, melatonin attenuated NaAsO2-evoked BTB disruption and sperm count reduction in adult mice. These results suggest that mitochondrial dysfunction-associated translational inhibition of barrier junction proteins is involved in As-mediated BTB disruption and sperm quality decline.

Accumulating data have demonstrated that long-term arsenic (As) exposure reduces testicular testosterone (T) synthesis. This study investigated the contribution of Leydig cell ferroptosis to As-impaired testicular T synthesis. Adult male C57BL/6J mice received NaAsO2 (0, 1.5, or 15 mg/L) by drinking water. As-exposed mice exhibited suppressed serum and testicular T levels with concomitant downregulation of T synthases. The number of Leydig cells, as determined by histopathology, immunohistochemistry, and immunofluorescence, was reduced in As-exposed mouse testes. Transcriptomic profiling identified ferroptosis as a significantly enriched pathway among differentially expressed genes (DEGs). Free ferrous ions were increased, and MDA and 4-HNE, two markers of lipid peroxidation, were elevated in As-exposed mouse testes. ACSL4 and NCOA4, two initiators of ferroptosis, were upregulated, and GPX4, a key ferroptosis repressor, was diminished in As-exposed mouse testes. Liproxstatin-1 (Lip-1), a specific ferroptosis inhibitor, protected against As-induced testicular Leydig cell ferroptosis. Moreover, pretreatment with Lip-1 attenuated As-induced declined of T synthases in mouse testes. Accordingly, Lip-1 pretreatment reversed As-induced reduction of testicular T synthesis. These results suggest that As exposure reduces testicular T synthesis partially by evoking Leydig cell ferroptosis in mouse testes.