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Protein expression profiling in head fragments during planarian regeneration after amputation

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Abstract

Following amputation, a planarian tail fragment can regrow into a complete organism including a well-organized brain within about 2–3 weeks, thus restoring the structure and function to presurgical levels. Despite the enormous potential of these animals for regenerative medicine, our understanding of the exact mechanism of planarian regeneration is incomplete. To better understand the molecular nature of planarian head regeneration, we applied two-dimensional electrophoresis (2-DE)/matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF)/time-of-flight mass spectrometry (TOF MS) technique to analyze the dynamic proteomic expression profiles over the course of 6 to 168 h post-decapitation. This approach identified a total of 141 differentially expressed proteins, 47 of which exhibited exceptionally high fold changes (≥3-fold change). Of these, Rx protein, an important regulator of head and brain development, was considered to be closely related to planarian head regeneration because of its exceptional high expression almost throughout the time course of regeneration process. Functional annotation analysis classified the 141 proteins into eight categories: (1) signaling, (2) Ca2+ binding and translocation, (3) transcription and translation, (4) cytoskeleton, (5) metabolism, (6) cell protection, (7) tissue differentiation, and (8) cell cycle. Signaling pathway analysis indicated that Wnt1/Ca2+ signaling pathway was activated during head regeneration. Integrating the analyses of proteome expression profiling, functional annotation, and signaling pathway, amputation-induced head reformation requires some mechanisms to promote cell proliferation and differentiation, including differential regulation of proapoptotic and antiapoptotic proteins, and the regulation of proliferation and differentiation-related proteins. Importantly, Wnt1/Ca2+ signaling pathway upregulates Rx expression, finally facilitating the differentiation of neoblasts into various cell types. Taken together, our study demonstrated that proteomic analysis approach used by us is a powerful tool in understanding molecular process related to head regeneration of planarian.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 31401209) and National Basic Research 973 Pre-research Program of China (No. 2010CB534905).

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Authors and Affiliations

  1. Animal Science and Technology School, Henan University of Science and Technology, Luoyang, 471003, China

    Xiaoguang Chen

  2. Key Laboratory for Cell Differentiation Regulation, Henan Normal University, 46# East of Construction Road, Xinxiang, 453007, China

    Cunshuan Xu

  3. College of Life Science, Henan Normal University, 46# East of Construction Road, Xinxiang, 453007, China

    Cunshuan Xu

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  1. Xiaoguang Chen
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  2. Cunshuan Xu
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Correspondence to Cunshuan Xu.

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Communicated by David A. Weisblat

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Supplementary Table 1

The numbers of upregulated, downregulated and up/downregulated proteins involved in different functional categories. Note: “—” indicates the absence of up/downregulated proteins (DOC 90 kb)

Supplementary Table 2

Highly regulated proteins that are identified by this study in each functional categories. Note: “NC” represents that the expression abundance of protein is not detected. Negative value presents the expression abundance lower than the control. The asterisk represents that the expression change of protein is more than 4-fold when compared to the control. (DOC 119 kb)

Supplementary Table 3

Head regeneration-involved proteins that have been identified by other studies. (DOC 45 kb)

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Chen, X., Xu, C. Protein expression profiling in head fragments during planarian regeneration after amputation. Dev Genes Evol 225, 79–93 (2015). https://doi.org/10.1007/s00427-015-0494-3

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