中国实验动物学报2026，Vol.34Issue(5)：639-654,16.DOI:10.3969/j.issn.1005-4847.2026.05.002
跨膜转运蛋白zgc:158423功能缺失导致氧化磷酸化功能紊乱诱发脑出血
Loss-of-function in transmembrane transporter zgc:158423 disrupts oxidative phosphorylation to induce intracerebral hemorrhage
摘要
Abstract
Objective To explore the mechanism by which the SLC3A2 homologous gene zgc:158423 influences intracerebral hemorrhage in adult zebrafish.Methods Clustered regularly interspaced short palindromic repeats associated protein 9(CRISPR/Cas9)technology was employed to generate zgc:158423 zebrafish mutants.Vascular endothelial cell-labeled transgenic fish Tg(kdrl:eGFP)were crossed with zgc:158423 mutants.zgc:158423-/-;Tg(kdrl:eGFP)mutants were obtained through genotyping and screening.Confocal laser scanning microscopy was used to observe cerebral vascular development phenotypes in 4 and 10 d post fertilization(dpf)larvae.Brain tissue clearing and light-sheet imaging were employed to examine cerebral vascular development in adult homozygous mutants,with cerebral hemorrhage rates quantified across different age groups.Transcriptome sequencing and bioinformatics analysis were performed on whole-brain tissue from adult mutants,with reverse transcription quantitative polymerase chain reaction(RT-qPCR)used to detect related gene expression.Results The zebrafish gene zgc:158423 is a direct homolog of human.SLC3A2,exhibiting high sequence conservation across species.Bioinformatics analysis and in situ hybridization experiments confirmed that zgc:158423 is specifically highly expressed in brain vascular endothelial cells during wild-type zebrafish development.zgc:158423 homozygous mutants exhibited a brain hemorrhage phenotype at 5,7 and 9 months post fertilization(mpf)with significantly increased hemorrhage rates;leakage was significantly increased in the brain tissue of the 9 mpf mutant,indicating impaired blood-brain barrier function.Ribonucleic acid sequencing(RNA-Seq)result identified 2,148 differentially expressed genes,comprising 950 upregulated and 1,198 downregulated genes.After knocking out zgc:158423,the blood-brain barrier(BBB)-related gene set and wingless/integrated(WNT)signaling pathway-related genes were significantly affected.kyoto encyclopedia of genes and genomes(KEGG)metabolic pathway analysis indicated enrichment of differentially expressed genes primarily involved in oxidative phosphorylation signaling.RT-qPCR detection revealed significant downregulation of key pathway genes atp5l,atp5fld,cox8a,cox 17,and ndufc2,while cox 15 expression was upregulated,indicating disrupted oxidative phosphorylation signaling.There was a reduction in ATP,NAD+,and the NAD+/NADH ratio in mutant brain tissue,while NADH significantly increased,further indicating impaired oxidative phosphorylation.Conclusions Deletion of the zgc:158423 gene in zebrafish disrupts oxidative phosphorylation signaling,inducing cerebral hemorrhage in adult zebrafish.关键词
zgc:158423/SLC3A2/斑马鱼/基因敲除/脑出血/氧化磷酸化Key words
zgc:158423/SLC3A2/zebrafish/gene knockout/intracerebral hemorrhage/oxidative phosphorylation分类
生物科学引用本文复制引用
池涌鑫,马骏驰,李晓玉,宁国柱,张晶晶..跨膜转运蛋白zgc:158423功能缺失导致氧化磷酸化功能紊乱诱发脑出血[J].中国实验动物学报,2026,34(5):639-654,16.基金项目
国家自然科学基金(32222028,32470889),广东省本科高校教学质量与教学改革工程建设项目(粤教高函[2024]9号,粤教高函[2024]30号),2024年广东省大学生创新创业项目(S202410571068,S202410571079).Funded by National Natural Science Foundation of China(32222028,32470889),Guangdong Provincial Project for Teaching Quality and Teaching Reform in High School(Yue Jiao Gao Han[2024]No.9,Yue Jiao Gao Han[2024]No.30),2024 Guangdong Provincial College Students' Innovation and Entrepreneurship Training Program(S202410571068,S202410571079). （32222028,32470889）
