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キーワード:
Astrocytes; Ca2+ signaling; Genetically encoded calcium indicator; GECI; GCaMP6; Glia; rAAV gene transduction; Two-photon microscopy
要旨:
Decades of research have unraveled the complex functioning of neurons in the central nervous system. Our knowledge of the second main player of the brain – the non-excitable glial cells – clearly lags behind that of neurons. Pioneering work in the 1990s provided evidence that star-shaped glial cells – astrocytes – sense and modulate neuronal activity by intracellular Ca2+ signals. However, the precise roles of astrocytic Ca2+ signaling in brain physiology and pathophysiology are still highly controversial, largely due to technical limitations of previous Ca2+ imaging tools. With recent innovations in laser microscopy and engineering of molecular probes, the field of glioscience is undergoing a revolution. This chapter describes the application of multiphoton microscopy and genetically encoded fluorescent Ca2+ indicators to reveal astrocytic Ca2+ signals in acute brain slices and in vivo, both in anesthetized and in awake behaving animals.
