ResearchPad - limb-regeneration https://www.researchpad.co Default RSS Feed en-us © 2020 Newgen KnowledgeWorks <![CDATA[Active Notch signaling is required for arm regeneration in a brittle star]]> https://www.researchpad.co/article/elastic_article_7845 Cell signaling pathways play key roles in coordinating cellular events in development. The Notch signaling pathway is highly conserved across all multicellular animals and is known to coordinate a multitude of diverse cellular events, including proliferation, differentiation, fate specification, and cell death. Specific functions of the pathway are, however, highly context-dependent and are not well characterized in post-traumatic regeneration. Here, we use a small-molecule inhibitor of the pathway (DAPT) to demonstrate that Notch signaling is required for proper arm regeneration in the brittle star Ophioderma brevispina, a highly regenerative member of the phylum Echinodermata. We also employ a transcriptome-wide gene expression analysis (RNA-seq) to characterize the downstream genes controlled by the Notch pathway in the brittle star regeneration. We demonstrate that arm regeneration involves an extensive cross-talk between the Notch pathway and other cell signaling pathways. In the regrowing arm, Notch regulates the composition of the extracellular matrix, cell migration, proliferation, and apoptosis, as well as components of the innate immune response. We also show for the first time that Notch signaling regulates the activity of several transposable elements. Our data also suggests that one of the possible mechanisms through which Notch sustains its activity in the regenerating tissues is via suppression of Neuralized1.

]]>
<![CDATA[Limber Pine (Pinus flexilis James), a Flexible Generalist of Forest Communities in the Intermountain West]]> https://www.researchpad.co/article/5989dadaab0ee8fa60bb99a8

As forest communities continue to experience interactions between climate change and shifting disturbance regimes, there is an increased need to link ecological understanding to applied management. Limber pine (Pinus flexilis James.), an understudied species of western North America, has been documented to dominate harsh environments and thought to be competitively excluded from mesic environments. An observational study was conducted using the Forest Inventory and Analysis Database (FIAD) to test the competitive exclusion hypothesis across a broad elevational and geographic area within the Intermountain West, USA. We anticipated that competitive exclusion would result in limber pine’s absence from mid-elevation forest communities, creating a bi-modal distribution. Using the FIAD database, limber pine was observed to occur with 22 different overstory species, which represents a surprising number of the woody, overstory species commonly observed in the Intermountain West. There were no biologically significant relationships between measures of annual precipitation, annual temperature, or climatic indices (i.e. Ombrothermic Index) and limber pine dominance. Limber pine was observed to be a consistent component of forest communities across elevation classes. Of the plots that contained limber pine regeneration, nearly half did not have a live or dead limber pine in the overstory. However, limber pine regeneration was greater in plots with higher limber pine basal area and higher average annual precipitation. Our results suggest limber pine is an important habitat generalist, playing more than one functional role in forest communities. Generalists, like limber pine, may be increasingly important, as managers are challenged to build resistance and resilience to future conditions in western forests. Additional research is needed to understand how different silvicultural systems can be used to maintain multi-species forest communities.

]]>
<![CDATA[Mechanism of Action of Secreted Newt Anterior Gradient Protein]]> https://www.researchpad.co/article/5989db45ab0ee8fa60bd861b

Anterior gradient (AG) proteins have a thioredoxin fold and are targeted to the secretory pathway where they may act in the ER, as well as after secretion into the extracellular space. A newt member of the family (nAG) was previously identified as interacting with the GPI-anchored salamander-specific three-finger protein called Prod1. Expression of nAG has been implicated in the nerve dependence of limb regeneration in salamanders, and nAG acted as a growth factor for cultured newt limb blastemal (progenitor) cells, but the mechanism of action was not understood. Here we show that addition of a peptide antibody to Prod1 specifically inhibit the proliferation of blastema cells, suggesting that Prod1 acts as a cell surface receptor for secreted nAG, leading to S phase entry. Mutation of the single cysteine residue in the canonical active site of nAG to alanine or serine leads to protein degradation, but addition of residues at the C terminus stabilises the secreted protein. The mutation of the cysteine residue led to no detectable activity on S phase entry in cultured newt limb blastemal cells. In addition, our phylogenetic analyses have identified a new Caudata AG protein called AG4. A comparison of the AG proteins in a cell culture assay indicates that nAG secretion is significantly higher than AGR2 or AG4, suggesting that this property may vary in different members of the family.

]]>
<![CDATA[Diffusion tensor tractography reveals muscle reconnection during axolotl limb regeneration]]> https://www.researchpad.co/article/5989db52ab0ee8fa60bdc720

Axolotls have amazing ability to regenerate their lost limbs. Our previous works showed that after amputation the remnant muscle ends remained at their original location whilst sending satellite cells into the regenerating parts to develop into early muscle fibers in the late differentiation stage. The parental and the newly formed muscle fibers were not connected until very late stage. The present study used non-invasive diffusion tensor imaging (DTI) to monitor weekly axolotl upper arm muscles after amputation of their upper arms. DTI tractography showed that the regenerating muscle fibers became visible at 9-wpa (weeks post amputation), but a gap was observed between the regenerating and parental muscles. The gap was filled at 10-wpa, indicating reconnection of the fibers of both muscles. This was confirmed by histology. The DTI results indicate that 23% of the muscle fibers were reconnected at 10-wpa. In conclusion, DTI can be used to visualize axolotls’ skeletal muscles and the results of muscle reconnection were in accordance with our previous findings. This non-invasive technique will allow researchers to identify the timeframe in which muscle fiber reconnection takes place and thus enable the study of the mechanisms underlying this reconnection.

]]>
<![CDATA[A Tunable Silk Hydrogel Device for Studying Limb Regeneration in Adult Xenopus Laevis]]> https://www.researchpad.co/article/5989daedab0ee8fa60bc00db

In certain amphibian models limb regeneration can be promoted or inhibited by the local wound bed environment. This research introduces a device that can be utilized as an experimental tool to characterize the conditions that promotes limb regeneration in the adult frog (Xenopus laevis) model. In particular, this device was designed to manipulate the local wound environment via a hydrogel insert. Initial characterization of the hydrogel insert revealed that this interaction had a significant influence on mechanical forces to the animal, due to the contraction of the hydrogel. The material and mechanical properties of the hydrogel insert were a factor in the device design in relation to the comfort of the animal and the ability to effectively manipulate the amputation site. The tunable features of the hydrogel were important in determining the pro-regenerative effects in limb regeneration, which was measured by cartilage spike formation and quantified by micro-computed tomography. The hydrogel insert was a factor in the observed morphological outcomes following amputation. Future work will focus on characterizing and optimizing the device’s observed capability to manipulate biological pathways that are essential for limb regeneration. However, the present work provides a framework for the role of a hydrogel in the device and a path forward for more systematic studies.

]]>