PRESTO

2 ARTICLES PUBLISHED IN JoVE

Genetics

An Optogenetic Method to Control and Analyze Gene Expression Patterns in Cell-to-cell Interactions

Akihiro Isomura^1,2, Ryoichiro Kageyama^1,3,4,5

¹Institute for Frontier Life and Medical Sciences, Kyoto University, ²Japan Science and Technology Agency, PRESTO, ³Institute for Integrated Cell-Material Sciences, Kyoto University, ⁴Graduate School of Medicine, Kyoto University, ⁵Graduate School of Biostudies, Kyoto University

Here, we present a protocol to analyze cell-to-cell transfer of oscillatory information by optogenetic control and live monitoring of gene expression. This approach provides a unique platform to test a functional significance of dynamic gene expression programs in multicellular systems.

Neuroscience

Dissection of Local Ca²⁺ Signals in Cultured Cells by Membrane-targeted Ca²⁺ Indicators

Hiroko Bannai^1,2, Matsumi Hirose², Fumihiro Niwa^2,3, Katsuhiko Mikoshiba²

¹Japan Science and Technology Agency, PRESTO, ²Laboratory for Developmental Neurobiology, RIKEN Center for Brain Science, ³École Normale Supèrieure, Institut de Biologie de l'ENS (IBENS), Institut national de la santè et de la recherche mèdicale (INSERM), Centre national de la recherche scientifique (CNRS), École Normale Supèrieure, PSL Research University

Here we present a protocol for Ca²⁺ imaging in neurons and glial cells, which enables the dissection of Ca²⁺ signals at subcellular resolution. This process is applicable to all cell types that allow the expression of genetically encoded Ca²⁺ indicators.