Current Topics in Microbiology and Immunology, 2012
Many vertebrates have the amazing ability to regenerate all or portions of appendages including l... more Many vertebrates have the amazing ability to regenerate all or portions of appendages including limbs, tails, fins, and digits. Unfortunately, our understanding of the cellular and molecular basis of appendage regeneration is severely lacking. However, recent technological advances that facilitate the tracking of cell lineages in vivo through space and time are allowing us to address the unknowns of regeneration, such as characterizing the cells that contribute to regeneration and identifying the tissues these cells differentiate into during regeneration. Here, we describe the experiments and the surprisingly uniform results that have emerged across diverse vertebrate species when specific cell lineages have been tracked during vertebrate appendage regeneration. These investigations show that vertebrates, from zebrafish to salamanders to mammals, utilize a limited amount of cellular plasticity to regenerate missing appendages. The universal approach to appendage regeneration is not to generate pluripotent cells that then differentiate into the new organ, but instead to generate lineage-restricted cells that are propagated in a progenitor-like state. Lessons learned from these natural cases of complex tissue regeneration might inform regenerative medicine on the best approach for re-growing complex tissues.
Salamanders are capable of regenerating amputated limbs by generating a mass of lineage-restricte... more Salamanders are capable of regenerating amputated limbs by generating a mass of lineage-restricted cells called a blastema. Blastemas only generate structures distal to their origin unless treated with retinoic acid (RA), which results in proximodistal (PD) limb duplications. Little is known about the transcriptional network that regulates PD duplication. In this study, we target specific retinoic acid receptors (RARs) to either PD duplicate (RA treatment or RARγ agonist) or truncate (RARβ antagonist) regenerating limbs. RARE-EGFP reporter axolotls showed divergent reporter activity in limbs undergoing PD duplication versus truncation, suggesting differences in patterning and skeletal regeneration. Transcriptomics identified expression patterns that explain PD duplication, including upregulation of proximal homeobox gene expression and silencing of distal-associated genes, whereas limb truncation was associated with disrupted skeletal differentiation. RARβ antagonism in uninjured li...
The spiny mouse, Acomys cahirinus displays a unique wound healing ability with regeneration of al... more The spiny mouse, Acomys cahirinus displays a unique wound healing ability with regeneration of all skin components in a scar-free manner. To identify orchestrators of this regenerative response we have performed proteomic analyses of skin from Acomys and Mus musculus before and after wounding. Of the ~2000 proteins identified many are expressed at similar levels in Acomys and Mus, but there are significant differences. Following wounding in Mus the complement and coagulation cascades, PPAR signaling pathway and ECM-receptor interactions predominate. In Acomys, other pathways predominate including the Wnt, MAPK, the ribosome, proteasome, endocytosis and tight junction pathways. Notable among Acomys specific proteins are several ubiquitin-associated enzymes and kinases, whereas in Mus immuno-modulation proteins characteristic of inflammatory response are unique or more prominent. ECM proteins such as collagens are more highly expressed in Mus, but likely more important is the higher e...
The African Spiny Mouse (Acomys spp.) is a unique outbred mammal capable of full, scar-free skin ... more The African Spiny Mouse (Acomys spp.) is a unique outbred mammal capable of full, scar-free skin regeneration. In vivo, we have observed rapid reepithelialization and deposition of normal dermis in Acomys after wounding. Acomys skin also has a lower modulus and lower elastic energy storage than normal lab mice, Mus musculus. To see if the different in vivo mechanical microenvironments retained an effect on dermal cells and contributed to regenerative behavior, we examined isolated keratinocytes in response to physical wounding and fibroblasts in response to varying substrate stiffness. Classic mechanobiology paradigms suggest stiffer substrates will promote myofibroblast activation, but we do not see this in Acomys dermal fibroblasts (DFs). Though Mus DFs increase organization of α-smooth muscle actin (αSMA)-positive stress fibers as substrate stiffness increases, Acomys DFs assemble very few αSMA-positive stress fibers upon changes in substrate stiffness. Acomys DFs generate lower ...
Current Topics in Microbiology and Immunology, 2012
Many vertebrates have the amazing ability to regenerate all or portions of appendages including l... more Many vertebrates have the amazing ability to regenerate all or portions of appendages including limbs, tails, fins, and digits. Unfortunately, our understanding of the cellular and molecular basis of appendage regeneration is severely lacking. However, recent technological advances that facilitate the tracking of cell lineages in vivo through space and time are allowing us to address the unknowns of regeneration, such as characterizing the cells that contribute to regeneration and identifying the tissues these cells differentiate into during regeneration. Here, we describe the experiments and the surprisingly uniform results that have emerged across diverse vertebrate species when specific cell lineages have been tracked during vertebrate appendage regeneration. These investigations show that vertebrates, from zebrafish to salamanders to mammals, utilize a limited amount of cellular plasticity to regenerate missing appendages. The universal approach to appendage regeneration is not to generate pluripotent cells that then differentiate into the new organ, but instead to generate lineage-restricted cells that are propagated in a progenitor-like state. Lessons learned from these natural cases of complex tissue regeneration might inform regenerative medicine on the best approach for re-growing complex tissues.
Salamanders are capable of regenerating amputated limbs by generating a mass of lineage-restricte... more Salamanders are capable of regenerating amputated limbs by generating a mass of lineage-restricted cells called a blastema. Blastemas only generate structures distal to their origin unless treated with retinoic acid (RA), which results in proximodistal (PD) limb duplications. Little is known about the transcriptional network that regulates PD duplication. In this study, we target specific retinoic acid receptors (RARs) to either PD duplicate (RA treatment or RARγ agonist) or truncate (RARβ antagonist) regenerating limbs. RARE-EGFP reporter axolotls showed divergent reporter activity in limbs undergoing PD duplication versus truncation, suggesting differences in patterning and skeletal regeneration. Transcriptomics identified expression patterns that explain PD duplication, including upregulation of proximal homeobox gene expression and silencing of distal-associated genes, whereas limb truncation was associated with disrupted skeletal differentiation. RARβ antagonism in uninjured li...
The spiny mouse, Acomys cahirinus displays a unique wound healing ability with regeneration of al... more The spiny mouse, Acomys cahirinus displays a unique wound healing ability with regeneration of all skin components in a scar-free manner. To identify orchestrators of this regenerative response we have performed proteomic analyses of skin from Acomys and Mus musculus before and after wounding. Of the ~2000 proteins identified many are expressed at similar levels in Acomys and Mus, but there are significant differences. Following wounding in Mus the complement and coagulation cascades, PPAR signaling pathway and ECM-receptor interactions predominate. In Acomys, other pathways predominate including the Wnt, MAPK, the ribosome, proteasome, endocytosis and tight junction pathways. Notable among Acomys specific proteins are several ubiquitin-associated enzymes and kinases, whereas in Mus immuno-modulation proteins characteristic of inflammatory response are unique or more prominent. ECM proteins such as collagens are more highly expressed in Mus, but likely more important is the higher e...
The African Spiny Mouse (Acomys spp.) is a unique outbred mammal capable of full, scar-free skin ... more The African Spiny Mouse (Acomys spp.) is a unique outbred mammal capable of full, scar-free skin regeneration. In vivo, we have observed rapid reepithelialization and deposition of normal dermis in Acomys after wounding. Acomys skin also has a lower modulus and lower elastic energy storage than normal lab mice, Mus musculus. To see if the different in vivo mechanical microenvironments retained an effect on dermal cells and contributed to regenerative behavior, we examined isolated keratinocytes in response to physical wounding and fibroblasts in response to varying substrate stiffness. Classic mechanobiology paradigms suggest stiffer substrates will promote myofibroblast activation, but we do not see this in Acomys dermal fibroblasts (DFs). Though Mus DFs increase organization of α-smooth muscle actin (αSMA)-positive stress fibers as substrate stiffness increases, Acomys DFs assemble very few αSMA-positive stress fibers upon changes in substrate stiffness. Acomys DFs generate lower ...
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