Transcriptome Analysis of Neural Tissues in Mouse Models of Methotrexate-induced Neural Tube Defects under Folate Deficiency
Objective To investigate the gene functions and transcription factors that potentially influence the development of neural tissues by analyzing the changes of gene transcriptional expression in the neural tissue of fetal mice with neural tube defects(NTDs)in-duced by low folate feeding combined with methotrexate(MTX).Methods SPF-grade 6-8-week-old healthy female ICR mice were divided into two groups:the control group,which received a normal diet,and the experimental group,which received a low-folic acid diet.The mice were fed under a low folate diet for four weeks and conceived,on day 7.5 of conception;the experimental group was intra-peritoneally injected with MTX at a concentration of 1.5mg/kg.On day 9.5 of conception,the brain and spinal cord were collected from both groups.Total RNA was extracted from the tissues for RNA-seq analysis.The differentially expressed genes(DEGs)were identified using DESeq software.Bioinformatic approaches were used to analyse functions and transcription factors associated with the DEGs in the experimental and control groups.Results There were 939 and 879 DEGs in the brain and the spinal cord tissue for the experimental group and the control group,respectively.Gene Ontology(GO)functional analysis showed that DEGs of biological process(BP)were mainly enriched in cellular process,biological regulation,developmental process,and metabolic process;DEGs of cellular component(CC)were mainly concentrated in cellular and organelle-related components;DEGs of molecular function(MF)were primarily involved in protein binding,transcriptional regulatory activity,and molecular function regulators.The DEGs function in the spinal cord had consistent results in the brain.The differentially expressed transcription factors in the brain and spinal cord were focused on zf-C2H2,bHLH,Ho-meobox,and STAT families.Conclusion The combination of low folate and MTX can induce transcriptional alterations in the genome of fetal mouse neural tissues.The DEGs identified are primarily associated with neural cell development and transcriptional regulation.The transcriptional regulators influencing neural development are concentrated in the zf-C2H2,bHLH,Homeobox,and STAT families.
NETL
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