Department of Pharmacotherapy, Washington State University College of Pharmacy and Pharmaceutical Sciences, Spokane, WA, USA.
Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, USA.
Epilepsy Res. 2020 Oct;166:106395. doi: 10.1016/j.eplepsyres.2020.106395. Epub 2020 Jun 20.
Vigabatrin (VGB; γ-vinyl-GABA) is an antiepileptic drug that elevates CNS GABA via irreversible inactivation of the GABA catabolic enzyme GABA-transaminase. VGB's clinical utility, however, can be curtailed by peripheral visual field constriction (pVFC) and thinning of the retinal nerve fiber layer (RNFL). Earlier studies from our laboratory revealed disruptions of autophagy by VGB. Here, we tested the hypothesis that VGB administration to animals would reveal alterations of gene expression in VGB-treated retina that associated with autophagy. VGB (140 mg/kg/d; subcutaneous minipump) was continuously administered to mice (n = 6 each VGB/vehicle) for 12 days, after which animals were euthanized. Retina was isolated for transcriptome (RNAseq) analysis and further validation using qRT-PCR and immunohistochemistry (IHC). For 112 differentially expressed retinal genes (RNAseq), two databases (Gene Ontology; Kyoto Encyclopedia of Genes and Genomes) were used to identify genes associated with visual function. Twenty four genes were subjected to qRT-PCR validation, and five (Gb5, Bdnf, Cplx9, Crh, Sox9) revealed significant dysregulation. IHC of fixed retinas verified significant down-regulation of Gb5 in photoreceptor cells. All of these genes have been previously shown to play a role in retinal function/circuitry signaling. Minimal impact of VGB on retinal autophagic gene expression was observed. This is the first transcriptome analysis of retinal gene expression associated with VGB intake, highlighting potential novel molecular targets potentially related to VGB's well known ocular toxicity.
氨己烯酸(VGB;γ-乙烯基-GABA)是一种抗癫痫药物,通过不可逆地失活 GABA 分解代谢酶 GABA-转氨酶来提高中枢神经系统 GABA。然而,VGB 的临床应用可能会受到外周视野收缩(pVFC)和视网膜神经纤维层(RNFL)变薄的限制。我们实验室的早期研究表明 VGB 会破坏自噬。在这里,我们假设在给予动物 VGB 后,VGB 处理的视网膜中的基因表达变化会与自噬有关。VGB(140mg/kg/d;皮下微泵)连续给药 12 天(每组 6 只 VGB/载体),然后处死动物。分离视网膜进行转录组(RNAseq)分析,并使用 qRT-PCR 和免疫组织化学(IHC)进一步验证。对于 112 个差异表达的视网膜基因(RNAseq),使用两个数据库(基因本体论;京都基因与基因组百科全书)来识别与视觉功能相关的基因。对 24 个基因进行 qRT-PCR 验证,其中 5 个(Gb5、Bdnf、CpIx9、Crh、Sox9)显示出显著的失调。固定视网膜的 IHC 验证了光感受器细胞中 Gb5 的显著下调。所有这些基因以前都被证明在视网膜功能/回路信号中发挥作用。VGB 对视网膜自噬基因表达的影响很小。这是与 VGB 摄入相关的视网膜基因表达的首次转录组分析,突出了潜在的新分子靶点,这些靶点可能与 VGB 众所周知的眼部毒性有关。