Transcriptomic and proteomic analysis of Hemidactylus frenatus during initial stages of tail regeneration

Abstract

Epimorphic regeneration of appendages is a complex and complete phenomenon found in selected animals. Hemidactylus frenatus, house gecko has the remarkable ability to regenerate the tail tissue upon autotomy involving epimorphic regeneration mechanism. This study has identified and evaluated the molecular changes at gene and protein level during the initial stages, i.e., during the wound healing and repair mechanism initiation stage of tail regeneration. Based on next generation transcriptomics and De novo analysis the transcriptome library of the gecko tail tissue was generated. A total of 254 genes and 128 proteins were found to be associated with the regeneration of gecko tail tissue upon amputation at 1, 2 and 5-day post amputation (dpa) against control, 0-dpa through differential transcriptomic and proteomic analysis. To authenticate the expression analysis, 50 genes were further validated involving RTPCR. 327 genes/proteins identified and mapped from the study showed association for Protein kinase A signaling, Telomerase BAG2 signaling, paxillin signaling, VEGF signaling network pathways based on network pathway analysis. This study empanelled list of transcriptome, proteome and the list of genes/proteins associated with the tail regeneration.

Figure 1

(a) Heap map expression of Hemidactylus frenatus transcript during tail tissue regeneration. (b) Heat map expression of differentially expressed proteins which are associated with tail regeneration; (c) Heat map expression of various genes differentially regulated during regeneration based on RTPCR analysis. (d) Venn diagram for number genes identified from Transcriptomics, proteomics and Real time PCR based gene expression analysis; e. Batches of genes/proteins which are differentially expressed at 1, 2 and 5-dpa. Genes/proteins which are in the red/organge arrow indicates the upregulation and in the green arrow indicates the downregulation.

Figure 2

RTPCR analysis of differentially expressed transcripts. (a) RTPCR Gel image of all the 50 genes along with housekeeping gene for 0-dpa (control), 1-dpa, 2-dpa and 5-dpa. (b) Bar diagram of expression visualising the up and downregulation of the 50 genes.

Figure 3

(a) Canonical Pathway associated with the differentially expressed genes/proteins for tail regeneration; (b) Cell morphology and Embryonic development network pathway; (c) Cellular assembly and organization network pathway; (d) Organ and organismal development network pathway; (e) Skeletal and Muscular development network pathway.