Extracellular Vesicles in the Forebrain Display Reduced miR-346 and miR-331-3p in a Rat Model of Chronic Temporal Lobe Epilepsy.
Extracellular Vesicles in the Forebrain Display Reduced miR-346 and miR-331-3p in a Rat Model of Chronic Temporal Lobe Epilepsy.
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在慢性颞叶癫痫大鼠模型中，前脑细胞外囊泡显示 miR-346 和 miR-331-3p 减少。
DOI：
10.1007/s12035-019-01797-1
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发表时间：
2020-03
影响因子：
5.1
通讯作者：
Shetty AK
中科院分区：
文献类型：
--
作者：
Gitaí DLG;Dos Santos YDR;Upadhya R;Kodali M;Madhu LN;Shetty AK
最初的脑损伤，如癫痫持续状态（SE）后，发展为慢性颞叶癫痫（TLE）。我们研究了se诱导的慢性TLE建立后前脑细胞外囊泡（EVs） miRNA组成的变化。我们通过分级腹腔注射kainic酸诱导年轻Fischer 344大鼠SE，结果在SE后约3个月出现持续的自发性复发性癫痫发作。我们通过超离心方法从慢性癫痫大鼠和年龄匹配的naïve对照动物的整个前脑中分离ev，并进行miRNA测序研究，以了解慢性癫痫中前脑源性ev的miRNA组成变化。naïve和癫痫前脑的EVs均表现为球形或杯状形态，大小范围相当，CD63表达，但缺乏深层细胞标志物GM130的表达。然而，mirna测序研究表明，癫痫前脑ev中3种mirna （miR-187-5p、miR-346和miR-331-3p）下调，4种mirna （miR-490-5p、miR-376b-3p、miR-493-5p和miR-124-5p）上调，倍数变化范围为1.5 ~ 2.4 （p<0.0006； FDR<0.05）。通过使用geNorm和Normfinder软件，我们通过qRT-PCR研究发现miR-487和miR-221是测量癫痫前脑中发现的改变mirna的最佳内参基因组合。验证显示，只有miR-346和miR-331-3p在癫痫前脑EVs中显著下调。这些mirna的富集途径分析显示，与慢性癫痫的分子机制相关的信号通路过度代表，包括gaba -能（miR-346靶点）和mTOR （miR-331-3p靶点）系统。因此，在慢性癫痫中，神经细胞中两种mirna进入ev的包装发生了相当大的改变。对这两种mirna的功能研究可能揭示它们在TLE的病理生理和治疗中的作用。
An initial precipitating injury in the brain, such as after status epilepticus (SE), evolves into chronic temporal lobe epilepsy (TLE). We investigated changes in the miRNA composition of extracellular vesicles (EVs) in the forebrain after the establishment of SE-induced chronic TLE. We induced SE in young Fischer 344 rats through graded intraperitoneal injections of kainic acid, which resulted in consistent spontaneous recurrent seizures at ~3 months post-SE. We isolated EVs from the entire forebrain of chronically epileptic rats and age-matched naïve control animals through an ultracentrifugation method and performed miRNA-sequencing studies to discern changes in the miRNA composition of forebrain-derived EVs in chronic epilepsy. EVs from both naïve and epileptic forebrains displayed spherical or cup-shaped morphology, a comparable size-range, and CD63 expression but lacked the expression of a deep cellular marker GM130. However, miRNA-sequencing studies suggested downregulation of 3 miRNAs (miR-187-5p, miR-346 and miR-331-3p) and upregulation of 4 miRNAs (miR-490-5p, miR-376b-3p, miR-493-5p and miR-124-5p) in EVs from epileptic forebrains with fold changes ranging from 1.5 to 2.4 (p<0.0006; FDR<0.05). By using geNorm and Normfinder software, we identified miR-487 and miR-221 as the best combination of reference genes for measurement of altered miRNAs found in the epileptic forebrain through qRT-PCR studies. The validation revealed that only miR-346 and miR-331-3p were significantly downregulated in EVs from the epileptic forebrain. The enrichment pathway analysis of these miRNAs showed an overrepresentation of signaling pathways that are linked to molecular mechanisms underlying chronic epilepsy, including GABA-ergic (miR-346 targets) and mTOR (miR-331-3p targets) systems. Thus, the packaging of two miRNAs into EVs in neural cells is considerably altered in chronic epilepsy. Functional studies on these two miRNAs may uncover their role in the pathophysiology and treatment of TLE.
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影响因子：
2.9
作者：
Goes Gitai, Daniel Leite;Fachin, Ana Lucia;Paco-Larson, Maria Luisa
通讯作者：
Paco-Larson, Maria Luisa
影响因子：
3.7
作者：
Fiorenza, Anna;Lopez-Atalaya, Jose P.;Barco, Angel
通讯作者：
Barco, Angel
神经外泌体货物作为急性脑损伤生物标志物的诊断潜力。
影响因子：
5.3
作者：
Goetzl L;Merabova N;Darbinian N;Martirosyan D;Poletto E;Fugarolas K;Menkiti O
通讯作者：
Menkiti O
影响因子：
3.7
作者：
de Araújo MA;Marques TE;Taniele-Silva J;Souza FM;de Andrade TG;Garcia-Cairasco N;Paçó-Larson ML;Gitaí DL
通讯作者：
Gitaí DL
影响因子：
3.4
作者：
Bertoglio D;Amhaoul H;Van Eetveldt A;Houbrechts R;Van De Vijver S;Ali I;Dedeurwaerdere S
通讯作者：
Dedeurwaerdere S