Hadas Yoav, Etlin Alex, Falk Haya, Avraham Oshri, Kobiler Oren, Panet Amos, Lev-Tov Aharon, Klar Avihu
Department of Medical Neurobiology, IMRIC, Hebrew University Medical School, Jerusalem, Israel.
Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.
Nucleic Acids Res. 2014 Oct 29;42(19):e148. doi: 10.1093/nar/gku750. Epub 2014 Aug 21.
The genetic dissection of spinal circuits is an essential new means for understanding the neural basis of mammalian behavior. Molecular targeting of specific neuronal populations, a key instrument in the genetic dissection of neuronal circuits in the mouse model, is a complex and time-demanding process. Here we present a circuit-deciphering 'tool box' for fast, reliable and cheap genetic targeting of neuronal circuits in the developing spinal cord of the chick. We demonstrate targeting of motoneurons and spinal interneurons, mapping of axonal trajectories and synaptic targeting in both single and populations of spinal interneurons, and viral vector-mediated labeling of pre-motoneurons. We also demonstrate fluorescent imaging of the activity pattern of defined spinal neurons during rhythmic motor behavior, and assess the role of channel rhodopsin-targeted population of interneurons in rhythmic behavior using specific photoactivation.
脊髓回路的基因剖析是理解哺乳动物行为神经基础的一种重要新方法。对特定神经元群体进行分子靶向,这是小鼠模型中神经元回路基因剖析的关键手段,是一个复杂且耗时的过程。在此,我们展示了一个用于对鸡发育中的脊髓神经元回路进行快速、可靠且廉价的基因靶向的回路解析“工具箱”。我们展示了对运动神经元和脊髓中间神经元的靶向、轴突轨迹的映射以及单个和群体脊髓中间神经元的突触靶向,以及前运动神经元的病毒载体介导标记。我们还展示了在节律性运动行为期间特定脊髓神经元活动模式的荧光成像,并使用特定光激活评估靶向中间神经元群体的通道视紫红质在节律性行为中的作用。