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Imaging cell-type-specific dynamics of mRNAs in living mouse brain
Chiso Nwokafor1, Robert H Singer2, Hyungsik Lim3
1Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, United States; Department of Biology, Graduate Center of the City University of New York, New York, NY 10016, United States.
Abstract:
We describe a method for visualizing mRNAs in living mouse. Nascent transcripts and cytoplasmic mRNAs were labeled via lentiviral expression of MS2 coat protein (MCP) tagged with fluorescent protein (MCP-XFP) in knock-in mice whose β-actin mRNAs contained MCP binding stem loops (MBS). Then the mRNA molecules were imaged in the live cerebral cortex through an optical cranial window by intravital two-photon microscopy. By means of the controlled expression of MCP-XFP, single mRNA particles could be detected differentially in the nucleus and cytoplasm of a specific cell type. Consequently, this method is useful for investigating the cell-type-dependent dynamics of mRNAs underlying the structure and function of the brain.
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