Protein expression pattern and analysis of differentially expressed peptides during various stages of tail regeneration in Hemidactylus flaviviridis

Abstract

Epimorphic regeneration is a process allowing the animal to regain its lost structure which depends on the resident pluripotent stem cells as well as de-differentiation of existing cells to form multi-potent stem cells. Many studies have been done to understand the appendage regeneration mechanism. The animal model used since decades is an urodele amphibian the axolotl. However, this ability is also seen in some members of reptiles, mainly lizards which on autotomy of tail regain the same by forming a replica of its lost tail. Lizards being closer to mammals are of greater interest and cannot be neglected. Hence, a stage specific protein profiling was undertaken in order to find the peptides playing a major role in epimorphosis. 2-DGE being basic tool for creating a protein profile was used. With advent of newer modern technology, label-free analysis which uses MS/MS was also performed. The study reports the peptides involved in apoptosis, inflammation and ECM remodelling across the stages of lizard tail regeneration for the first time. Apart from these peptides, structural protein, enzymes involved in metabolism have also been highlighted in the current study to give a bigger picture of the processes and the specific peptides required for tail regeneration.

Salamander-like tail regeneration in the West African lungfish

“…RNA-seq analysis comparing tail versus limb blastema in the common wall lizard (Podaris muralis) revealed genes exclusively upregulated in the regenerating tail, which include those coding for growth factors (Wnts, Shh, Bmps, Fgfs) as well as genes within broad categories of ECM, inflammation and immunity, metabolism and cytoskeleton [23]. A similar molecular profile was observed in the regenerating tail proteome of the northern house gecko, Hemidactylus flaviviridis, which showed enrichment for peptides involved in immune response, ECM remodeling, Fgfs and Bmps [24]. In the green anole lizard (Anolis carolinensis), the distal tip of regenerating tail at 25 days post amputation (dpa) is enriched for genes that fall in gene ontology (GO) categories of wound response, immune response, hormonal regulation and embryonic morphogenesis [25].…”

Inherent variations in the cellular events at the site of amputation orchestrate scar‐free wound healing in the tail and scarred wound healing in the limb of lizardHemidactylus flaviviridis

“…Time points for tissue collection were decided based on the discrete events associated with wound healing in tail and in limb. These have been characterized for the tail through the course of various studies in our lab . It has been observed through histology in our lab that following autotomy, hemostasis is achieved rapidly and a scab is formed as early as 1dpa.…”

“…These have been characterized for the tail through the course of various studies in our lab. 13 It has been observed through histology in our lab that following autotomy, hemostasis is achieved rapidly and a scab is formed as early as 1dpa. Subsequently, a thin layer of epithelium covers the wound surface on 2dpa.…”

“…(B) Shows the scarred wound healing where reepithelialization starts along with fibroblast proliferation, deposition of collagen (shown by arrowheads) and ECM development to form the granulation tissue. Brown dotted line represents the hemostasis phase, red dotted line shows the inflammation phase, blue dotted line is for epidermis covering the wound and green dotted line depicts the proliferation of epidermis in scar‐free wound healing while in scarred wound healing it represents proliferation of fibroblast cells and scab formation (Adapted from Murawala et al and Diegelmann and Evans). [Color figure can be viewed at wileyonlinelibrary.com]…”

Transcriptome and Proteome analysis ofHemidactylus frenatusduring initial stages of tail regeneration

“…This also might pave in giving insights into spinal injuries and the fabrication of replacement therapies [11]. Research on the regeneration of tail and its governing molecular mechanism in other lizard species like Hemidactylus falviviridis [12]; Green Anole [13][14]; and other vertebrates axolotl [15][16][17] have been carried out related to histology, proteomics and genomics [12 -17].…”