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Whited, Jessica

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Whited

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Jessica

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Whited, Jessica
Author's Publications: search.filters.isAuthorOfPublication.d06e19a7-5779-4d8a-a6d2-d498ab7dc632

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    Identification of regenerative roadblocks via repeat deployment of limb regeneration in axolotls
    (Nature Publishing Group UK, 2017) Bryant, Donald M.; Sousounis, Konstantinos; Payzin-Dogru, Duygu; Bryant, Sevara; Sandoval, Aaron Gabriel W.; Martinez Fernandez, Jose; Mariano, Rachelle; Oshiro, Rachel; Wong, Alan Y.; Leigh, Nicholas; Johnson, Kimberly; Whited, Jessica
    Axolotl salamanders are powerful models for understanding how regeneration of complex body parts can be achieved, whereas mammals are severely limited in this ability. Factors that promote normal axolotl regeneration can be examined in mammals to determine if they exhibit altered activity in this context. Furthermore, factors prohibiting axolotl regeneration can offer key insight into the mechanisms present in regeneration-incompetent species. We sought to determine if we could experimentally compromise the axolotl’s ability to regenerate limbs and, if so, discover the molecular changes that might underlie their inability to regenerate. We found that repeated limb amputation severely compromised axolotls’ ability to initiate limb regeneration. Using RNA-seq, we observed that a majority of differentially expressed transcripts were hyperactivated in limbs compromised by repeated amputation, suggesting that mis-regulation of these genes antagonizes regeneration. To confirm our findings, we additionally assayed the role of amphiregulin, an EGF-like ligand, which is aberrantly upregulated in compromised animals. During normal limb regeneration, amphiregulin is expressed by the early wound epidermis, and mis-expressing this factor lead to thickened wound epithelium, delayed initiation of regeneration, and severe regenerative defects. Collectively, our results suggest that repeatedly amputated limbs may undergo a persistent wound healing response, which interferes with their ability to initiate the regenerative program. These findings have important implications for human regenerative medicine.
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    TALEN‐mediated gene editing of the thrombospondin‐1 locus in axolotl
    (John Wiley and Sons Inc., 2015) Kuo, Tzu‐Hsing; Kowalko, Johanna E.; DiTommaso, Tia; Nyambi, Mandi; Montoro, Daniel; Essner, Jeffrey J.; Whited, Jessica
    Abstract Loss‐of‐function genetics provides strong evidence for a gene's function in a wild‐type context. In many model systems, this approach has been invaluable for discovering the function of genes in diverse biological processes. Axolotls are urodele amphibians (salamanders) with astonishing regenerative abilities, capable of regenerating entire limbs, portions of the tail (including spinal cord), heart, and brain into adulthood. With their relatively short generation time among salamanders, they offer an outstanding opportunity to interrogate natural mechanisms for appendage and organ regeneration provided that the tools are developed to address these long‐standing questions. Here we demonstrate targeted modification of the thrombospondin‐1 (tsp‐1) locus using transcription‐activator‐like effector nucleases (TALENs) and identify a role of tsp‐1 in recruitment of myeloid cells during limb regeneration. We find that while tsp‐1‐edited mosaic animals still regenerate limbs, they exhibit a reduced subepidermal collagen layer in limbs and an increased number of myeloid cells within blastemas. This work presents a protocol for generating and genotyping mosaic axolotls with TALEN‐mediated gene edits.
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    Regeneration Review Reprise
    (BioMed Central, 2010) Whited, Jessica; Tabin, Clifford
    There have been notable advances in the scientific understanding of regeneration within the past year alone, including two recently published in BMC Biology. Increasingly, progress in the regeneration field is being inspired by comparisons with stem cell biology and enabled by newly developed techniques that allow simultaneous examination of thousands of genes and proteins. See research articles http://www.biomedcentral.com/1741-7007/7/83 and http://www.biomedcentral.com/1741-7007/8/5.
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    Limb regeneration revisited
    (BioMed Central, 2009) Whited, Jessica; Tabin, Clifford
    The investigation of vertebrate limb regeneration, a favorite topic of early developmental biologists, is enjoying a renaissance thanks to recently developed molecular and genetic tools, as indicated in recent papers in BMC Biology and BMC Developmental Biology. Classical experiments provide a rich context for interpreting modern functional studies.