Figure 2. olMG cells react preferentially to PRC injuries by apical migration.

(A–B’’’) In vivo imaging of hatchling rx2::H2B-eGFP medaka retinae which were either injured in the ONL or the ganglion cell layer (GCL) (asterisks) using a two-photon laser and imaged consecutively until 30 hpi (n > 10 fish each, data obtained from >10 independent experiments each). (A–A’’’) After PRC injuries olMG nuclei (arrowheads) start migrating apically towards the ONL layer from 17 hpi on. The migration is not coordinated among different migrating nuclei. (B–B’’’) After RGC injuries no migration of olMG nuclei can be detected until 30 hpi. Scale bars are 10 μm. (C–D’’) In vivo imaging of hatchling rx2::H2B-eGFP medaka retinae which were either injured in the ONL or the GCL (asterisks) using a two-photon laser and imaged every second day after injury (n > 10 fish each, data obtained from >10 independent experiments each). (C–C’’) PRC injuries result in an apical migration of olMG nuclei into the injury site. The following days until 6 dpi the nuclei do not migrate back toward the INL resulting in a gap of olMG nuclei in the INL. (D–D’’) After RGC injuries no migration of olMG nuclei can be detected until 6 dpi. Scale bars are 10 μm.

Figure 2—figure supplement 1. Two-photon mediated laser ablation enables targeted cell ablation in the retina resulting in specific cell death signatures.

(A–B) In vivo imaging of hatchling rx2::H2B-eGFP medaka retinae which were either injured in the ONL or the GCL (asterisks) using a two-photon laser. Targeted cell type ablation can be achieved; PRCs (A) as well as cells of the GCL (B) can be ablated. (C–D) Cryosections of hatchling medaka retinae which were either injured in the ONL or the GCL and fixed 16 hpi. TUNEL stainings (magenta) to detect programmed cell death show specific cell death of either ONL (C) or GCL (D). Scale bars are 10 μm.

Figure 2—figure supplement 2. Increased RGC injuries lead to swelling and secondary cell death in the PRC layer.

(A–B) In vivo imaging of hatchling rx2::H2B-eGFP medaka retinae which were either injured in the GCL (asterisks) using a two-photon laser and imaged 2 days later (n = 10 fish, data obtained from five independent experiments). Large RGC injuries induce swelling and cell death in the PRC layer (arrowheads). olMG nuclei are largely depleted from the INL. Scale bars are 10 µm.

Figure 2—video 1. In vivo imaging of olMG nuclei reactions to a PRC injury.

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DOI: 10.7554/eLife.32319.011

Figure 2—video 2. In vivo imaging of olMG nuclei reactions to a RGC injury.

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DOI: 10.7554/eLife.32319.012