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Folic acid alleviates X-ray irradiation-induced jaw malformation in zebrafish

ABSTRACT: X-rays are a form of ionizing radiation that has sufficient energy to remove electrons from atoms, thereby creating potentially harmful ions. X-ray irradiation during organogenesis can have profound detrimental effects, depending on the developmental stage and irradiating energy. In this study, we examined the impact of X-ray irradiation on zebrafish development at 36 h post-fertilization and observed marked jaw malformation, which was alleviated by folic acid pretreatment. Mechanistic studies revealed that folic acid pretreatment suppressed irradiation-induced production of reactive oxygen species. Transcriptome analysis performed at 24 h and 48 h post-irradiation revealed dysregulated expression of apolipoprotein A-IV a (apoa4a), methionine adenosyltransferase 1A (mat1a), heat shock protein 90 alpha class A member 1 tandem duplicate 1 (hsp90aa1.1), FKBP prolyl isomerase 5 (fkbp5), plac8 onzin related protein 3 (ponzr3), and prostaglandin E synthase 3a (ptges3a), which was not observed in zebrafish treated with folic acid-treated before the irradiation. These findings suggest that folic acid alleviates X-ray irradiation-induced jaw malformation in zebrafish, at least in part, by reducing oxidative stress and ameliorating dysregulated gene expression.

ORGANISM(S): Danio rerio

PROVIDER: GSE334519 | GEO | 2026/07/08

REPOSITORIES: GEO

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Project description:Exposure to environmental contaminants can disrupt normal development of the early vertebrate skeleton. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) impairs craniofacial skeletal development across many vertebrate species and its effects are especially prominent in early life stages of fish. TCDD activates the aryl hydrocarbon receptor (AHR), a transcription factor that mediates most if not all TCDD responses. We investigated the transcriptional response in the developing zebrafish jaw following TCDD exposure using DNA microarrays. Zebrafish larvae were exposed to TCDD at 96 h postfertilization (hpf) and jaw cartilage tissue was harvested for microarray analysis at 1, 2, 4 and 12 h postexposure (hpe). Numerous chondrogenic transcripts were misregulated by TCDD in the jaw. Comparison of transcripts altered by TCDD in jaw with transcripts altered in embryonic heart showed that the transcriptional responses in the jaw and the heart were strikingly different. Sox9b, a critical chondrogenic transcription factor, was the most significantly reduced transcript in the jaw. We hypothesized that the TCDD reduction of sox9b expression plays an integral role in affecting formation of the embryonic jaw. Morpholino knock down of sox9b expression demonstrated that partial reduction of sox9b expression alone was sufficient to produce a TCDD-like jaw phenotype. Heterozygous sox9b deletion mutant embryos were sensitized to TCDD. Lastly, embryos injected with sox9b mRNA and then exposed to TCDD blocked TCDD-induced jaw toxicity in approximately 14% of sox9b-injected embryos. These results suggest that reduced sox9b expression in TCDD-exposed zebrafish embryos contributes to jaw malformation. Experiment Overall Design: Three independent replicate microarray time course experiments were performed comparing transcript levels between TCDD-exposed and control zebrafish. For each experiment, zebrafish were exposed to TCDD for 1 h starting at 96 hpf as described above. For each time point (97, 98, 100 and 108 hpf) and treatment jaw samples were pooled from 10 dissections for RNA isolation and hybridization with Affymetrix zebrafish arrays (Affymetrix, Santa Clara, CA). Each microarray contains roughly 14,900 probes corresponding to approximately 30% of the zebrafish genome. For each array, total RNA (1 Âµg) was isolated from 10 jaw microdissections with the QIAGEN RNeasy Mini kit following the manufacturerâs protocol (QIAGEN, Valencia, CA). The One-Cycle Target Labeling and Control Reagents kit was used to synthesize cDNA and biotinylated cRNA following the manufacturerâs protocol (Affymetrix, Santa Clara, CA). Biotin-labeled cRNA (15 Âµg) was fragmented and hybridized onto Affymetrix Zebrafish Genechip Arrays following the protocol in the Affymetrix Genechip Expression Analysis Technical Manual. Following hybdrization, the arrays were washed and stained with streptavidin-phycoerythrin on an Affymetrix Fluidics Station 400 using the protocol EukGE WS2v4. Arrays were scanned with an Agilent Gene Array Scanner.

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Folic acid alleviates X-ray irradiation-induced jaw malformation in zebrafish

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