Optogenetic Interrogation of ChR2-Expressing GABAergic Interneurons After Transplantation into the Mouse Brain
Optogenetic Interrogation of ChR2-Expressing GABAergic Interneurons After Transplantation into the Mouse Brain
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DOI：
10.1007/978-1-0716-0830-2_15
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发表时间：
2021-01-01
期刊：
影响因子：
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通讯作者：
Naegele, Janice R.
中科院分区：
文献类型：
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作者：
Arshad, Muhammad N.;Aaron, Gloster B.;Naegele, Janice R.
本文介绍了研究方法，以调查移植的GABA能中间神经元和内源性神经元之间的突触连接在成年小鼠海马的发展。我们的方案强调了逆转录病毒标记成人出生的GC的方法，这是成人大脑中为数不多的在整个生命中不断更新的细胞类型之一。通过逆转录病毒的精确靶向，成年出生的GC的标记可以与移植细胞的光遗传学刺激和脑切片中的电生理学相结合，以测试GABA能中间神经元是否与宿主脑神经元整合并建立抑制性突触连接。成人神经发生的改变是TLE和癫痫发作发展和严重程度的重要因素。当与逆转录病毒标记相结合时，我们在本章中描述的方法可以用来确定移植是否改变了成年神经发生的过程或海马的其他特性。这些方法有助于确定用于难治性癫痫患者的潜在细胞替代疗法的参数。
This paper describes research methods to investigate the development of synaptic connections between transplanted GABAergic interneurons and endogenous neurons in the adult mouse hippocampus. Our protocol highlights methods for retroviral labeling adult-born GCs, one of the few cell types in the adult brain to be continuously renewed throughout life. By precise targeting of the retrovirus, labeling of adult-born GCs can be combined with optogenetic stimulation of the transplanted cells and electrophysiology in brain slices, to test whether the GABAergic interneurons integrate and establish inhibitory synaptic connections with host brain neurons. Modifications to adult neurogenesis are an important contributing factor in the development and severity of TLE and seizures. When combined with retroviral labeling, the approaches we describe in this chapter can be used to determine whether transplantation modifies the process of adult neurogenesis or other properties of the hippocampus. These approaches are helping to define parameters for potential cell replacement therapies to be used in patients with intractable seizure disorders.