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. 2016 Jun 1;525(1):65–78. doi: 10.1002/cne.24042

Figure 4.

Figure 4

Birthdating Analysis by photoconverted fluorescent Protein Tracing In vivo, with Subpopulation Markers (BAPTISM) method (Caron et al., 2008) for identifying the time of terminal differentiation (birthdate) of motoneurons in Tg(huC:Kaede; isl1:GFP) zebrafish, which express photolabile fluorescent protein Kaede pan‐neuronally. (A) Neurons born by the time of initial photoconversion (one of five timepoints shown) contain converted Kaede in initial image, taken at 5 dpf. Second photoconversion converts remaining Kaede, leaving GFP as the only green signal in Final image. Images are compared to determine birthdate of GFP+ motoneurons. (B) Dorsal plane (z12, dashed line) from a 5 dpf Tg(huC:Kaede; isl1:GFP) larva initially photoconverted at 36 hpf. Yellow arrowheads indicate an nIII motoneuron (isl:GFP+, final image) born by 36 hpf (converted huC:Kaede+, initial image). White arrowheads indicate a neuron born by 36 hpf not belonging to nIII/nIV. Orange arrowheads indicate an nIV motoneuron not born by 36 hpf. Scale bars = 20 μm.