Figure 4

Nucleic acid metabolism and cell division dynamics in live rat hippocampal slices with ethynyl uridine (EU) and ethynyl deoxyuridine (EdU).

(a) Newly synthesized RNA is visualized in the nucleoli of CA1 pyramidal neurons. Rat hippocampal slices are incubated with 2 mM EU (alkyne 2123 cm⁻¹) for 7 hours before SRS imaging. (b) Newly synthesized DNA is imaged inside the nucleus of neuron at the subgranular zone of dentate gyrus, indicating neurogenesis. Rat hippocampal slices are incubated with 300 μM EdU (alkyne 2123 cm⁻¹) for 3 days before SRS imaging. (a,b) Scale bar: 10 μm. (c) Different DNA incorporation patterns are observed with two-color pulse-chase labeling of EdU, likely from two cell division cycles. Rat hippocampal slices are pulsed with 400 μM ^12-12EdU (alkyne 2123 cm⁻¹) for 12 hours then chased with 400 μM ^13-13EdU (alkyne 2048 cm⁻¹) for 14 hours before SRS imaging. (d) Symmetric neural progenitor cell division is captured in the dentate gyrus of hippocampal tissue. Inset: zoom-in area showing the distribution of newly synthesized DNA. Rat hippocampal slices are incubated with 200 μM EdU for 3 days before SRS imaging. (c,d) Scale bar: 20 μm.