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MicroRNA profiling of antler stem cells in potentiated and dormant states and their potential roles in antler regeneration

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Abstract

MicroRNAs (miRNAs) can effectively regulate gene expression at the post-transcriptional level and play a critical role in tissue growth, development and regeneration. Our previous studies showed that antler regeneration is a stem cell-based process and antler stem cells reside in the periosteum of a pedicle, the permanent bony protuberance, from which antler regeneration takes place. Antlers are the only mammalian organ that can fully regenerate and hence provide a unique opportunity to identify miRNAs that are involved in organ regeneration. In the present study, we used next generation sequencing technology sequenced miRNAs of the stem cells derived from either the potentiated or the dormant pedicle periosteum. A population of both conserved and 20 deer-specific miRNAs was identified. These conserved miRNAs were derived from 453 homologous hairpin precursors across 88 animal species, and were further grouped into 167 miRNA families. Among them, the miR-296 is embryonic stem cell-specific. The potentiation process resulted in the significant regulation (>±2 Fold, q value <0.05) of conserved miRNAs; 8 miRNA transcripts were down- and 6 up-regulated. Several GO biology processes and the Wnt, MAPK and TGF-beta signaling pathways were found to be up-regulated as part of antlerogenic stem cell potentiation process. This research has identified miRNAs that are associated either with the dormant or the potentiated antler stem cells and identified some target miRNAs for further research into their role played in mammalian organ regeneration.

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Acknowledgments

The authors would like to thank staff from Antler Research Group of Institute of Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, for the help of pedicle periosteum collection. We wish to thank Dr Dawn Coates for reading through the paper and giving valuable comments. This work was partially funded by Chinese National 863 program (2011AA100603), partially funded by Chinese National Science Foundation (31070878, 31402035) and partially Jilin Merit Aid Study Abroad Programs (2015-23). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences, Jilin, 130112, People’s Republic of China

    Hengxing Ba, Datao Wang & Chunyi Li

  2. State Key Laboratory for Molecular Biology of Special Economic Animals, Jilin, 130112, People’s Republic of China

    Hengxing Ba, Datao Wang & Chunyi Li

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  1. Hengxing Ba
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  2. Datao Wang
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Correspondence to Chunyi Li.

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The authors declare that they have no competing interests.

Ethical approval

The deer were euthanized and slaughtered in strict accordance with the local slaughtering procedure approved by the Animal Ethics Committee of Institute of Special Wild Economic Animals and Plants, Chinese Academy of Agricultural Sciences.

Additional information

Communicated by S. Hohmann.

Electronic supplementary material

Below is the link to the electronic supplementary material.

438_2015_1158_MOESM1_ESM.pdf

Figure S1. Status of antler regeneration. Pedicle stumps just prior to the hard antlers (button) to cast, which is evidenced by the shiny appearance at the distal end of the pedicle skin (arrow) (PDF 370 kb)

438_2015_1158_MOESM2_ESM.pdf

Figure S2. Distribution of the annotation of assigned small RNAs in the PPP and the DPP libraries. A) Unique small RNAs. B) Total small RNAs (PDF 165 kb)

438_2015_1158_MOESM3_ESM.pdf

Figure S3. Validation of 20 novel miRNA expression in the PP by stem-loop RT-PCR. Length of the stem-loop RT-PCR primers are around 40 bp (PDF 1108 kb)

Table S1. Primer sequences used for six selected differentially expressed miRNAs (XLSX 9 kb)

Table S2. Primer sequences used for 20 novel deer-specific miRNAs (XLSX 10 kb)

Table S3. miRNAs differentially expressed in the PPP and the DPP (XLSX 167 kb)

438_2015_1158_MOESM7_ESM.txt

Table S4. Predicted target gene results (PPP-T) from the down-regulated miRNAs in the PPP through TargetScan (TXT 192 kb)

Table S5. Predicted target gene results (DPP-T) from the up-regulated miRNAs in the PPP through TargetScan (TXT 372 kb)

Table S6. Results of GO biological process analysis to the target genes in the PPP-T (TXT 19 kb)

Table S7. Results of GO biological process analysis to the target genes in the DPP-T (TXT 5 kb)

Supplementary material 11 (TXT 96 kb)

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Ba, H., Wang, D. & Li, C. MicroRNA profiling of antler stem cells in potentiated and dormant states and their potential roles in antler regeneration. Mol Genet Genomics 291, 943–955 (2016). https://doi.org/10.1007/s00438-015-1158-8

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