Azoxystrobin Impairs Neuronal Migration and Induces ROS Dependent Apoptosis in Cortical Neurons

Abstract

Fungicides often cause genotoxic stress and neurodevelopmental disorders such as autism (ASD). Fungicide-azoxystrobin (AZOX) showed acute and chronic toxicity to various organisms, and remained a concern for ill effects in developing neurons. We evaluated the neurotoxicity of AZOX in developing mouse brains, and observed prenatal exposure to AZOX reduced neuronal viability, neurite outgrowth, and cortical migration process in developing brains. The 50% inhibitory concentration (IC50) of AZOX for acute (24 h) and chronic (7 days) exposures were 30 and 10 μM, respectively. Loss in viability was due to the accumulation of reactive oxygen species (ROS), and inhibited neurite outgrowth was due to the deactivation of mTORC1 kinase activity. Pretreatment with ROS scavenger- N-acetylcysteine (NAC) reserved the viability loss and forced activation of mTORC1 kinase revived the neurite outgrowth in AZOX treated neurons. Intra-amniotic injection of AZOX coupled with in utero electroporation of GFP-labelled plasmid in E15.5 mouse was performed and 20 mg/kg AZOX inhibited radial neuronal migration. Moreover, the accumulation of mitochondria was significantly reduced in AZOX treated primary neurons, indicative of mitochondrial deactivation and induction of apoptosis, which was quantified by Bcl2/Bax ratio and caspase 3 cleavage assay. This study elucidated the neurotoxicity of AZOX and explained the possible cure from it.

Azoxystrobin Disrupts Colonic Barrier Function in Mice via Metabolic Disorders Mediated by Gut Microbiota

“…7 An in vitro study showed that AZO could impair neuronal migration and induce ROS-dependent apoptosis in cortical neurons. 8 In conclusion, AZO residues are increasingly recognized as a global public health issue.…”

Azoxystrobin exposure impairs meiotic maturation by disturbing spindle formation in mouse oocytes

“…Therefore, the effect of AZO on oocyte maturation was first examined by evaluating the GVBD and PBE rates. Previous studies have shown that AZO at 1 μM was sufficient to induce changes in transcriptional activity, while 5-10 μM could result in reactive oxygen species accumulation in primary neuronal cells (Pearson et al, 2016;Simon et al, 2019;Kang et al, 2021). Moreover, AZO at micromolar concentrations (1-3 μM) has been shown to modify membrane ion permeability in thymocytes (Imura et al, 2019).…”

“…Accumulating evidence suggests that AZO has the potential to induce developmental toxicity in animals. For instance, AZO has been reported to impair cortical migration by increasing reactive oxygen species (ROS) levels and inducing mitochondrial deactivation in mouse embryos (Kang et al, 2021). Also, parental exposure to AZO in zebrafish may cause mortality and developmental malformations in their F1 offspring (Cao et al, 2019).…”

Azoxystrobin induces apoptosis via PI3K/AKT and MAPK signal pathways in oral leukoplakia progression

“…And no reproductive or offspring effects were seen in the 2-generation rat reproduction study. However, it was shown that AZOX reduced neuronal viability, neurite outgrowth, and cortical migration process in developing brains ( Kang et al, 2021 ). Studies have shown that AZOX can target mitochondria and inhibit the electron transport of the oxidative phosphorylation process, leading to reduced cellular energy ( Huttemann et al, 2007 ; Shi et al, 2017 ).…”

“…The relevant study has shown that AZOX inhibits the viability of human esophageal cancer cells KYSE-150 and induces mitochondrial pathway of apoptosis ( Shi et al, 2017 ). And AZOX inhibits neuronal migration and induces apoptosis in cortical neurons ( Kang et al, 2021 ). In our previous study, we found that AZOX inhibited oral carcinogenesis by specifically targeting mitochondrial complex III, leading to ROS accumulation, MMP reduction and apoptosis.…”