Directing astroglia from the cerebral cortex into subtype specific functional neurons.
Directing astroglia from the cerebral cortex into subtype specific functional neurons.
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DOI：
10.1371/journal.pbio.1000373
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发表时间：
2010-05-18
期刊：
影响因子：
9.8
通讯作者：
Berninger B
中科院分区：
文献类型：
--
作者：
Heinrich C;Blum R;Gascón S;Masserdotti G;Tripathi P;Sánchez R;Tiedt S;Schroeder T;Götz M;Berninger B
单一转录因子的强制表达可以选择性和稳定地将培养的星形胶质细胞转化为突触形成的兴奋性和抑制性神经元。来自出生后大脑皮层的星形胶质细胞可以在体外重编程以在神经源性转录因子的强制表达后产生神经元，从而为内源性星形胶质细胞用于脑修复的潜在用途开辟了新的途径。然而，在以前的尝试中，星形胶质细胞衍生的神经元未能建立功能性突触，这严重限制了功能性神经发生。因此，它仍然是未知的，是否可以通过选择性表达不同的神经源性命运决定因素，对不同的神经元亚型的身份来自重编程星形胶质细胞。在这里，我们表明，由沉默抗性逆转录病毒载体驱动的神经源性命运决定因素的强烈和持续表达指示体外出生后皮层的星形胶质细胞成熟为功能齐全的突触形成神经元。重要的是，星形胶质细胞衍生的神经元的神经递质命运选择可以通过不同的神经源性转录因子的选择性表达来控制：背侧端脑命运决定因子neurogenin-2（Neurog 2）的强制表达指导皮质星形胶质细胞产生形成突触的神经元能神经元;相反，腹侧端脑命运决定子Dlx 2诱导GABA能特性，尽管与Neurog 2相比，Dlx 2介导的神经元重编程的总体效率低得多，这表明皮质星形胶质细胞具有更高的能力来响应背侧端脑命运决定子。然而，有趣的是，当在用Dlx 2转导之前首先将星形胶质细胞扩增为神经球细胞时，大大促进了星形胶质细胞朝向GABA能神经元的产生的重编程。重要的是，这种在神经球条件下的扩增和随后用不同的神经源性转录因子重编程的方法也可以扩展到从成人损伤的大脑皮层分离的反应性星形胶质细胞，从而允许选择性地产生谷氨酸能或GABA能神经元。这些数据提供的证据表明，皮质星形胶质细胞可以通过强迫表达单一的神经原性转录因子，稳定地产生完全分化的神经元，经历跨细胞谱系的转换。此外，星形胶质细胞的神经元重编程不限于出生后阶段，但也可以实现从终末分化的星形胶质细胞的成年大脑皮层损伤诱导的再激活。大脑由两种主要的细胞类型组成：神经元和神经胶质细胞，神经元传递信息，神经胶质细胞支持和保护神经元。有趣的是，有证据表明，一些神经胶质细胞，包括星形胶质细胞，可以通过特定的蛋白质直接转化为神经元，这种转化可能有助于受损脑组织的功能修复。然而，为了使修复的大脑区域正常运作，重要的是将星形胶质细胞定向为适当的神经元亚类。在这项研究中，我们表明，从大脑皮层的非神经源性星形胶质细胞可以在体外重新编程，只使用一个单一的转录因子，以产生功能齐全的兴奋性或抑制性神经元。我们通过强迫表达相同的转录因子来实现这一结果，这些转录因子在胚胎前脑发育过程中指导这些不同神经元亚型的发生。此外，我们证明，反应性星形胶质细胞从成人皮质局部损伤后，可以重新编程成突触形成兴奋性或抑制性神经元以下类似的策略。我们的研究结果提供的证据表明，内源性神经胶质细胞可能被证明是一个有前途的战略，取代神经元已退化，由于创伤或疾病。
Forced expression of single defined transcription factors can selectively and stably convert cultured astroglia into synapse-forming excitatory and inhibitory neurons. Astroglia from the postnatal cerebral cortex can be reprogrammed in vitro to generate neurons following forced expression of neurogenic transcription factors, thus opening new avenues towards a potential use of endogenous astroglia for brain repair. However, in previous attempts astroglia-derived neurons failed to establish functional synapses, a severe limitation towards functional neurogenesis. It remained therefore also unknown whether neurons derived from reprogrammed astroglia could be directed towards distinct neuronal subtype identities by selective expression of distinct neurogenic fate determinants. Here we show that strong and persistent expression of neurogenic fate determinants driven by silencing-resistant retroviral vectors instructs astroglia from the postnatal cortex in vitro to mature into fully functional, synapse-forming neurons. Importantly, the neurotransmitter fate choice of astroglia-derived neurons can be controlled by selective expression of distinct neurogenic transcription factors: forced expression of the dorsal telencephalic fate determinant neurogenin-2 (Neurog2) directs cortical astroglia to generate synapse-forming glutamatergic neurons; in contrast, the ventral telencephalic fate determinant Dlx2 induces a GABAergic identity, although the overall efficiency of Dlx2-mediated neuronal reprogramming is much lower compared to Neurog2, suggesting that cortical astroglia possess a higher competence to respond to the dorsal telencephalic fate determinant. Interestingly, however, reprogramming of astroglia towards the generation of GABAergic neurons was greatly facilitated when the astroglial cells were first expanded as neurosphere cells prior to transduction with Dlx2. Importantly, this approach of expansion under neurosphere conditions and subsequent reprogramming with distinct neurogenic transcription factors can also be extended to reactive astroglia isolated from the adult injured cerebral cortex, allowing for the selective generation of glutamatergic or GABAergic neurons. These data provide evidence that cortical astroglia can undergo a conversion across cell lineages by forced expression of a single neurogenic transcription factor, stably generating fully differentiated neurons. Moreover, neuronal reprogramming of astroglia is not restricted to postnatal stages but can also be achieved from terminally differentiated astroglia of the adult cerebral cortex following injury-induced reactivation. The brain consists of two major cell types: neurons, which transmit information, and glial cells, which support and protect neurons. Interestingly, evidence suggests that some glial cells, including astroglia, can be directly converted into neurons by specific proteins, a transformation that may aid in the functional repair of damaged brain tissue. However, in order for the repaired brain areas to function properly, it is important that astroglia be directed into appropriate neuronal subclasses. In this study, we show that non-neurogenic astroglia from the cerebral cortex can be reprogrammed in vitro using just a single transcription factor to yield fully functional excitatory or inhibitory neurons. We achieved this result through forced expression of the same transcription factors that instruct the genesis of these distinct neuronal subtypes during embryonic forebrain development. Moreover we demonstrate that reactive astroglia isolated from the adult cortex after local injury can be reprogrammed into synapse-forming excitatory or inhibitory neurons following a similar strategy. Our findings provide evidence that endogenous glial cells may prove a promising strategy for replacing neurons that have degenerated due to trauma or disease.
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DOI：
10.1073/pnas.0711961105
发表时间：
2008-04-22
影响因子：
11.1
作者：
Feng, Ru;Desbordes, Sabrina C.;Graf, Thomas
通讯作者：
Graf, Thomas
影响因子：
5.3
作者：
Brill, Monika S.;Snapyan, Marina;Goetz, Magdalena
通讯作者：
Goetz, Magdalena
影响因子：
64.8
作者：
Fitzsimonds, RM;Song, HJ;Poo, MM
通讯作者：
Poo, MM
影响因子：
3.5
作者：
Hack, MA;Sugimori, M;Götz, M
通讯作者：
Götz, M
DOI：
10.1523/jneurosci.4516-08.2009
发表时间：
2009-01-28
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Fagel DM;Ganat Y;Cheng E;Silbereis J;Ohkubo Y;Ment LR;Vaccarino FM
通讯作者：
Vaccarino FM