Monitoring Population Membrane Potential Signals During Functional Development of Neuronal Circuits in Vertebrate Embryos
The functional organization of the vertebrate central nervous system (CNS) during the early phase of development has long been unclear because conventional electrophysiological means have several technical limitations. First, early embryonic neurons are small and fragile, and the application of microelectrodes is often difficult. Second, the simultaneous recording of electrical activity from multiple sites is limited, and as a consequence, response patterns of neural networks cannot be assessed. Optical recording techniques with voltage-sensitive dyes have overcome these obstacles and provided a new approach to the analysis of the functional development/organization of the CNS. In this chapter, we provide detailed information concerning the recording of optical signals in the embryonic nervous system. After outlining methodological considerations, we present examples of recent progress in optical studies on the embryonic nervous system wfith special emphasis on two topics. The first is the study of how synapse networks form in specific neuronal circuits. The second is the study of nonspecific correlated wave activity, which is considered to play a fundamental role in neural development. These studies clearly demonstrate the utility of fast voltage-sensitive dye imaging as a powerful tool for elucidating the functional organization of the vertebrate embryonic CNS.
KeywordsOptical Signal Correlate Activity Optical Recording Intrinsic Signal Transmitted Light Intensity
We thank Joel C. Glover, Akihiko Hirota, Tetsuro Sakai, Hitoshi Komuro, Yusuke Katoh, Yang Xue-Song, Yoshiyasu Arai, Hiraku Mochida, Itaru Yazawa, Shinichi Sasaki, Toshihisa Tanaka, Naohisa Miyakawa, and Masae Kinoshita for their contribution in the experiments. We express our gratitude to Drs. Lowrence B. Cohen and Brian M. Salzberg for discussions throughout the course of our work and critical reading of the manuscript. We also thank Dr. Shigeo Yasui and Hayashibara Biochemical Laboratories/Kankoh-Shikiso Kenkyusho for synthesizing many dyes including NK2761 at our request. The present study was supported by grants from the Ministry of Education-Science-Culture of Japan and the HFSP, and research funds from the Astellas Foundation for Research on Metabolic Disorders.
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