Targeted two-photon chemical apoptotic ablation of defined cell types in vivo
Robert A Hill1,2, Eyiyemisi C Damisah1,3, Fuyi Chen1,2
1Department of Neurology, Yale School of Medicine, New Haven, Connecticut 06511, USA.
Abstract:
A major bottleneck limiting understanding of mechanisms and consequences of cell death in complex organisms is the inability to induce and visualize this process with spatial and temporal precision in living animals. Here we report a technique termed two-photon chemical apoptotic targeted ablation (2Phatal) that uses focal illumination with a femtosecond-pulsed laser to bleach a nucleic acid-binding dye causing dose-dependent apoptosis of individual cells without collateral damage. Using 2Phatal, we achieve precise ablation of distinct populations of neurons, glia and pericytes in the mouse brain and in zebrafish. When combined with organelle-targeted fluorescent proteins and biosensors, we uncover previously unrecognized cell-type differences in patterns of apoptosis and associated dynamics of ribosomal disassembly, calcium overload and mitochondrial fission. 2Phatal provides a powerful and rapidly adoptable platform to investigate in vivo functional consequences and neural plasticity following cell death as well as apoptosis, cell clearance and tissue remodelling in diverse organs and species.
Insights
Scientists developed a new method, two-photon chemical apoptotic targeted ablation (2Phatal), to precisely induce and visualize cell death in living animals. This technique allows for detailed study of apoptosis and its effects on tissue remodeling and neural plasticity.
Area of Science:
- Cell Biology
- Neuroscience
- Biotechnology
Background:
- Understanding cell death mechanisms in vivo is crucial for studying diseases and tissue repair.
- Current methods lack the spatial and temporal precision needed to study apoptosis in living organisms.
Purpose of the Study:
- To develop a novel technique for precise induction and visualization of apoptosis in vivo.
- To investigate cell-type-specific differences in apoptotic processes and their consequences.
Main Methods:
- Developed two-photon chemical apoptotic targeted ablation (2Phatal) using a femtosecond-pulsed laser to induce dose-dependent apoptosis.
- Applied 2Phatal to precisely ablate specific cell populations (neurons, glia, pericytes) in mouse brains and zebrafish.
- Combined 2Phatal with organelle-targeted fluorescent proteins and biosensors to visualize cellular dynamics during apoptosis.
Main Results:
- Achieved precise, dose-dependent apoptosis of individual cells without collateral damage.
- Uncovered previously unrecognized cell-type differences in apoptotic patterns.
- Observed distinct dynamics of ribosomal disassembly, calcium overload, and mitochondrial fission during apoptosis.
Conclusions:
- 2Phatal is a powerful and adaptable platform for in vivo apoptosis research.
- Enables investigation of functional consequences of cell death, neural plasticity, and tissue remodeling.
- Facilitates studies across diverse organs and species.
