BackgroundSalamanders regenerate their limbs after amputation. However, the molecular mechanism of this unique regeneration remains unclear. In this study, isobaric tags for relative and absolute quantification (iTRAQ) coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS) was employed to quantitatively identify differentially expressed proteins in regenerating limbs 3, 7, 14, 30 and 42 days post amputation (dpa).ResultsOf 2636 proteins detected in total, 253 proteins were differentially expressed during different regeneration stages. Among these proteins, Asporin, Cadherin-13, Keratin, Collagen alpha-1(XI) and Titin were down-regulated. CAPG, Coronin-1A, AnnexinA1, Cathepsin B were up-regulated compared with the control. The identified proteins were further analyzed to obtain information about their expression patterns and functions in limb regeneration. Functional analysis indicated that the differentially expressed proteins were associated with wound healing, immune response, cellular process, metabolism and binding.ConclusionsThis work indicated that significant proteome alternations occurred during salamander limb regeneration. The results may provide fundamental knowledge to understand the mechanism of limb regeneration.Electronic supplementary materialThe online version of this article (10.1186/s12864-017-4125-4) contains supplementary material, which is available to authorized users.

Salamanders completely regenerate their limbs after amputation. Thus, these animals are unique models to investigate the mechanisms modulating regeneration in vertebrates. To investigate the influence of microRNAs (miRNAs) on newt limb regeneration, the miRNAs and mRNAs were simultaneously profiled using Illumina HiSeq 2500 System during limb regeneration of Cynops orientalis at 3, 7, 14, 30 and 42 days postamputation. A total of 203 miRNAs and 4230 mRNAs were identified to be differentially expressed. Together with the proteomic data obtained from our previous study, integrative analysis of multiple profiling data sets was performed to construct an interaction network of differentially expressed miRNAs, mRNAs and proteins. Results of GO and KEGG analyses showed that the differentially expressed miRNA targets were mainly directed to cytoskeletal remodeling and carbohydrate metabolism. The stage-specific regulation of miRNAs on their targets was analyzed by hierarchical clustering analysis and validated by qRT-PCR. The negative regulation of miR-223 and miR-133a on their targets was tested by performing dual luciferase reporter assay. The integration analysis will provide a powerful tool to identify the regulatory mechanisms of miRNAs and their targets. The results may have implications in understanding the complex mechanisms underlying newt limb regeneration.

ABSTRACT. In China, Moschus berezovskii (forest musk deer), a firstclass national protected animal, was earlier widely distributed. However, wild populations of the forest musk deer have declined because of human activity and habitat loss. In order to gather useful information for its conservation and management, we investigated the genetic diversity and population structure of this species by analyzing a 632-bp fragment of mitochondrial DNA (mtDNA) control region in three wild populations in Shaanxi Province, China. The average A+T content (64.1%) of mtDNA was higher than that of G+C (35.9%). A total of 178 variable sites (about 28.03% of the total nucleotides in the sequence) were detected in 71 individuals. The nucleotide diversity (P I ) in the 71 individuals was 0.04688, and the average nucleotide differences (K) were 21.238. The 71 individuals belonged to 33 haplotypes according to the determined sequences. The average genetic distance (P) among the haplotypes of the species was 0.169. The phylogenetic tree constructed using the neighbor-joining method revealed that these individuals were clustered into three groups, but the individual distribution in those groups was disordered. These data indicated the variation and rich genetic diversity in the three populations of