Retinal Degeneration and Regeneration in Xenopus laevis Tadpoles

Xenopus laevis offers unique advantages for studying retinal regeneration due to its strong regenerative capacity and amenability to transgenesis and genome editing. We present a detailed protocol for inducing and monitoring rod photoreceptor degeneration and regeneration in Tg(rho:GFP-NTR) tadpoles, where nitroreductase expression under the rhodopsin promoter enables conditional and specific ablation of rods upon metronidazole exposure. In addition, we describe the generation of albino Tg(rho:GFP-NTR); tyr ^-/- tadpoles using CRISPR/Cas9-mediated knockout of the tyrosinase gene, providing an alternative to existing albino Tg(rho:GFP-NTR) lines and facilitating real-time fluorescence imaging by reducing pigmentation interference. Finally, we detail methods to assess Müller glial proliferation (BrdU/Sox9 colabeling) and rod neurogenesis (BrdU pulse-chase). This manuscript provides a robust experimental model that enables the investigation of the cellular and molecular mechanisms underlying retinal regeneration in X. laevis.