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Biology subjects

Oonk, K.

Publications and source records attributed to Oonk, K..

2 recordsLinked to original sources

Asymmetric regeneration dynamics in zebrafish pectoral fins

Appendage amputation in regeneration-capable systems results in the full re-formation of lost tissue and cell populations. Despite an appreciation of this phenomenon in a select number of organisms, there is still much to quantitatively understand about the tissue growth dynamics during limb regeneration. To address these gaps in knowledge, we developed novel methods that enable longitudinal, quantitative imaging of zebrafish pectoral fin regeneration and uncovered a persistent spatial asymmetry in growth dynamics between anterior and posterior regions of the regenerating fin. Spatiotemporally resolved transcriptomics suggests differential anteroposterior regulation of the Mitogen-activated protein kinase (MAPK) and Wnt signaling pathways in pectoral fins. This asymmetry was also detected using quantitative imaging of biosensors reporting canonical Wnt and Erk pathway activity. In addition to known static spatially asymmetric genes, bulk transcriptomic analysis uncovered a large subset of genes with temporally asymmetric peaks in expression levels between anterior and posterior regions of the fin across regeneration timepoints, suggesting a global differential regulation of the regenerative program. Taken together, our results establish pectoral fins as a platform for quantitative analyses of appendage regeneration, and they identify persistent spatial asymmetries in growth, gene expression, and signaling activity during regeneration.

cell biology↗

Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

Tissue regeneration enhancer elements (TREEs) direct expression of target genes in injured and regenerating tissues. Additionally, TREEs of zebrafish origin were shown to direct the expression of transgenes in border zone regions after cardiac injury when packaged into recombinant AAV vectors and introduced into mice. Future implementation of TREEs into AAV-based vectors as research tools and potential gene therapy modalities will require a more detailed understanding of expression dynamics and potential off-target effects. Here, we applied in vivo bioluminescent imaging to mice systemically injected with AAV vectors containing different combinations of capsids, enhancers, and timing of delivery. Longitudinal tracking of TREE-based expression over time revealed distinct amplitudes and durations of reporter gene expression in the injured heart directed by different TREEs. The liver-de-targeted AAV capsid, AAV.cc84, was able to deliver TREEs either pre- or post-cardiac injury to negate off-target expression in the liver while maintaining transduction in the heart. By screening AAV9-based capsid libraries dosed systemically in mice post-cardiac injury, we discovered a new capsid variant, AAV.IR41, with enhanced transduction at cardiac border zone regions, and with elevated transduction of TREE driven transgenes versus conventional AAV9 vectors. In vivo bioluminescence imaging offers insights into how enhancers and engineered capsids can be implemented to modulate spatiotemporal transgene expression for future targeted therapies.

developmental biology↗