M. W. El-Saadi, L. Rivers, Xinli Tian, Hong Sun, Xiao-Hong Lu
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Using two-photon imaging and 3-D reconstruction, we have illustrated the robust neurodegeneration of cortex pyramidal neuron, striatal medium spiny neuron and caught in action the engulfment of neurons and blood vessels by microglia. Moreover, we've documented dramatic axon degradation consistent with Wallerian pathology while the cell bodies remain intact (\"dying back\"), opening the possibility of therapeutic intervention. We also observed unexpected Tyrosine Hydroxylase (TH) positive cell bodies in the striatum, suggesting the brain responds to neurodegeneration by expressing these TH cells in an attempt to compensate for dopaminergic denervation. Most notably, we've developed a novel genetic method to visualize neuron morphology and gain a more accurate understanding of the cellular mechanisms underlying neurodegeneration. MACT represents a powerful genetics method to explore therapeutic interventions, including opto-and chemo-genetics in neurodegenerative diseases in vivo.","PeriodicalId":196856,"journal":{"name":"2016 32nd Southern Biomedical Engineering Conference (SBEC)","volume":"31 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2016-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"MACT (Mosaicism with AAV Mediated Conditional Transgenesis) for Single Neuron Analysis of Neurodegeneration in Vivo, a Proof of Principle in Focal Cerebral Ischemia\",\"authors\":\"M. W. El-Saadi, L. Rivers, Xinli Tian, Hong Sun, Xiao-Hong Lu\",\"doi\":\"10.1109/SBEC.2016.65\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The World Health Organization (WHO) warns neurodegenerative diseases (ND) in our aging population will sharply increase over the coming decades. Developing biological tools is paramount in the search for novel therapies to stop the onset or halt the progression of ND. We have developed a novel genetic method MACT (Mosaicism with AAV mediated Conditional Transgenesis) for single neuron analysis. MACT integrates the retrograde labelling capability of AAV and a conditional genetic reporter mouse model for sparse genetic labelling to reveal detailed morphology of different cellular types. As proof of principle, we subsequently subjected MACT mice to a middle cerebral artery occlusion (MCAO) leading to extensive neurodegeneration. Using two-photon imaging and 3-D reconstruction, we have illustrated the robust neurodegeneration of cortex pyramidal neuron, striatal medium spiny neuron and caught in action the engulfment of neurons and blood vessels by microglia. Moreover, we've documented dramatic axon degradation consistent with Wallerian pathology while the cell bodies remain intact (\\\"dying back\\\"), opening the possibility of therapeutic intervention. We also observed unexpected Tyrosine Hydroxylase (TH) positive cell bodies in the striatum, suggesting the brain responds to neurodegeneration by expressing these TH cells in an attempt to compensate for dopaminergic denervation. Most notably, we've developed a novel genetic method to visualize neuron morphology and gain a more accurate understanding of the cellular mechanisms underlying neurodegeneration. 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引用次数: 0
摘要
世界卫生组织(WHO)警告说,在未来几十年里,老龄人口中神经退行性疾病(ND)的发病率将急剧上升。开发生物工具对于寻找新的治疗方法来阻止ND的发生或进展至关重要。我们开发了一种新的遗传方法MACT (Mosaicism with AAV介导的条件转化),用于单个神经元的分析。MACT结合AAV逆行标记能力和条件遗传报告小鼠模型进行稀疏遗传标记,揭示不同细胞类型的详细形态。作为原理证明,我们随后对MACT小鼠进行大脑中动脉闭塞(MCAO)导致广泛的神经变性。利用双光子成像和三维重建技术,我们展示了皮层锥体神经元、纹状体中棘神经元的强大神经变性,以及小胶质细胞对神经元和血管的吞噬。此外,我们已经记录了与沃勒氏病理一致的显著轴突退化,而细胞体保持完整(“死亡”),开启了治疗干预的可能性。我们还在纹状体中观察到意想不到的酪氨酸羟化酶(TH)阳性细胞体,这表明大脑通过表达这些TH细胞来补偿多巴胺能失神经支配,从而对神经变性做出反应。最值得注意的是,我们开发了一种新的遗传方法来可视化神经元形态,并更准确地了解神经变性的细胞机制。MACT代表了一种强大的遗传学方法来探索治疗干预,包括体内神经退行性疾病的光遗传学和化学遗传学。
MACT (Mosaicism with AAV Mediated Conditional Transgenesis) for Single Neuron Analysis of Neurodegeneration in Vivo, a Proof of Principle in Focal Cerebral Ischemia
The World Health Organization (WHO) warns neurodegenerative diseases (ND) in our aging population will sharply increase over the coming decades. Developing biological tools is paramount in the search for novel therapies to stop the onset or halt the progression of ND. We have developed a novel genetic method MACT (Mosaicism with AAV mediated Conditional Transgenesis) for single neuron analysis. MACT integrates the retrograde labelling capability of AAV and a conditional genetic reporter mouse model for sparse genetic labelling to reveal detailed morphology of different cellular types. As proof of principle, we subsequently subjected MACT mice to a middle cerebral artery occlusion (MCAO) leading to extensive neurodegeneration. Using two-photon imaging and 3-D reconstruction, we have illustrated the robust neurodegeneration of cortex pyramidal neuron, striatal medium spiny neuron and caught in action the engulfment of neurons and blood vessels by microglia. Moreover, we've documented dramatic axon degradation consistent with Wallerian pathology while the cell bodies remain intact ("dying back"), opening the possibility of therapeutic intervention. We also observed unexpected Tyrosine Hydroxylase (TH) positive cell bodies in the striatum, suggesting the brain responds to neurodegeneration by expressing these TH cells in an attempt to compensate for dopaminergic denervation. Most notably, we've developed a novel genetic method to visualize neuron morphology and gain a more accurate understanding of the cellular mechanisms underlying neurodegeneration. MACT represents a powerful genetics method to explore therapeutic interventions, including opto-and chemo-genetics in neurodegenerative diseases in vivo.