Funkční studium iniciace regenerace ocasu embryí Xenopus laevis

Netušil, Jiří

The recent introduction of high-throughput sequencing techniques rapidly changes our perception of vertebrate regeneration. One of the model organisms intensely studied for its regenerative potential is the embryonic tadpole stage of African clawed frog (Xenopus laevis). This thesis aims to review and functionally validate some of the latest findings gained by RNA sequencing of regenerating Xenopus laevis tail. We specifically focus on gene expression changes during the early phases of regeneration and how their absence affects the progression and phenotypic outcome of this process. Our analysis confirms that Regeneration initiating cells (RICs) identified by single-cell and spatial RNA sequencing represent a vital element in successful tail regeneration. The RIC marker genes mmp9 and pmepa1 affect the processes linked to extracellular matrix remodelling and the migration pattern of previously described Regeneration organizing cells (ROCs) while not interfering with the myeloid cell lineage. We further demonstrate a novel use of Vivo Morpholino oligonucleotides in a transient knockdown assay and offer an assessment of its benefits and limitations. This thesis highlights the importance of spatio-temporal regulation happening on both cellular and molecular levels during the initial 24 hours of...

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