Differentiation and functional incorporation of embryonic stem cell-derived GABAergic interneurons in the dentate gyrus of mice with temporal lobe epilepsy.
Differentiation and functional incorporation of embryonic stem cell-derived GABAergic interneurons in the dentate gyrus of mice with temporal lobe epilepsy.
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DOI：
10.1523/jneurosci.2683-11.2012
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发表时间：
2012-01-04
期刊：
影响因子：
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通讯作者：
Naegele JR
中科院分区：
文献类型：
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作者：
Maisano X;Litvina E;Tagliatela S;Aaron GB;Grabel LB;Naegele JR
神经系统疾病的细胞治疗需要对疾病相关的神经病理学和产生用于移植的神经元的程序有广泛的了解。在许多严重的获得性颞叶癫痫（TLE）患者中，齿状回表现为硬化和gaba能中间神经元变性。越来越多的证据表明，通过将胎儿gaba能祖细胞移植到患有TLE的啮齿动物的齿状回中可以获得治疗效果，但人类胎儿细胞的稀缺性限制了其在患者群体中的适用性。相比之下，几乎无限数量的神经祖细胞可以从胚胎干细胞（ES）中获得。基于ES细胞的TLE神经修复疗法需要移植神经元功能整合并取代退化的细胞类型的证据。为了解决这些问题，我们将具有腹侧前脑特征的小鼠胚胎干细胞来源的神经祖细胞（ESNPs）移植到TLE小鼠齿状回门部，并评估移植神经元的分化、苔藓纤维发芽、细胞形态和电生理特性。此外，我们比较了移植神经元与内源性肝门间神经元的电生理特性。大多数移植ESNPs分化为表达钙结合蛋白parvalbumin、calbindin或calretinin的gaba能神经元间亚型。没有观察到苔藓纤维发芽的整体抑制，然而，esnp衍生的神经元在内分子层和整个门叶形成密集的轴突树突。移植神经元的全细胞海马切片电生理记录和形态学分析确定了5种基本类型；大多数具有强烈的后超极化和光滑或稀疏的棘状树突形态，类似内源性海马中间神经元。此外，自发兴奋性突触后电流的细胞内记录表明，新细胞在功能上整合到癫痫海马回路中。
Cell therapies for neurological disorders require an extensive knowledge of disease-associated neuropathology and procedures for generating neurons for transplantation. In many patients with severe acquired temporal lobe epilepsy (TLE), the dentate gyrus exhibits sclerosis and GABAergic interneuron degeneration. Mounting evidence suggests that therapeutic benefits can be obtained by transplanting fetal GABAergic progenitors into the dentate gyrus in rodents with TLE, but the scarcity of human fetal cells limits applicability in patient populations. By contrast, virtually limitless quantities of neural progenitors can be obtained from embryonic stem (ES) cells. ES cell-based therapies for neurological repair in TLE require evidence that the transplanted neurons integrate functionally and replace cell types that degenerate. To address these issues, we transplanted mouse ES cell-derived neural progenitors (ESNPs) with ventral forebrain identities into the hilus of the dentate gyrus of mice with TLE and evaluated graft differentiation, mossy fiber sprouting, cellular morphology and electrophysiological properties of the transplanted neurons. Additionally we compared electrophysiological properties of the transplanted neurons to endogenous hilar interneurons in mice without TLE. The majority of transplanted ESNPs differentiated into GABAergic interneuron subtypes expressing calcium-binding proteins parvalbumin, calbindin or calretinin. Global suppression of mossy fiber sprouting was not observed, however, ESNP-derived neurons formed dense axonal arborizations in the inner molecular layer and throughout the hilus. Whole-cell hippocampal slice electrophysiological recordings and morphological analyses of the transplanted neurons identified five basic types; most with strong after-hyperpolarizations and smooth or sparsely spiny dendritic morphologies resembling endogenous hippocampal interneurons. Moreover, intracellular recordings of spontaneous excitatory postsynaptic currents indicated that the new cells functionally integrate into epileptic hippocampal circuitry.