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Planarian myosin essential light chain is involved in the formation of brain lateral branches during regeneration

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Abstract

The myosin essential light chain (ELC) is a structure component of the actomyosin cross-bridge, however, the functions in the central nervous system (CNS) development and regeneration remain poorly understood. Planarian Dugesia japonica has revealed fundamental mechanisms and unique aspects of neuroscience and neuroregeneration. In this study, the cDNA DjElc, encoding a planarian essential light chain of myosin, was identified from the planarian Dugesia japonica cDNA library. It encodes a deduced protein with highly conserved functionally domains EF-Hand and Ca2+ binding sites that shares significant similarity with other members of ELC. Whole mount in situ hybridization studies show that DjElc expressed in CNS during embryonic development and regeneration of adult planarians. Loss of function of DjElc by RNA interference during planarian regeneration inhibits brain lateral branches regeneration completely. In conclusion, these results demonstrated that DjElc is required for maintenance of neurons and neurite outgrowth, particularly for involving the brain later branch regeneration.

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Acknowledgments

This work was supported by grants from Shandong Province Natural Science Foundation of China (ZR2013CM011) and State Key Laboratory of Genetic Resource and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences (GREKF09-04) and a Project of Shandong Province Higher Educational Science and Technology Program, China (J11LC09).

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    Authors and Affiliations

    1. Laboratory of Developmental and Evolutionary Biology, School of Life Sciences, Shandong University of Technology, 12 Zhangzhou Road, Zibo, 255049, People’s Republic of China

      Shuying Yu, Xuhui Chen, Zuoqing Yuan, Luming Zhou, Qiuxiang Pang & Bosheng Zhao

    2. State Key Laboratory of Genetic Resource and Evolution, Kunming Institute of Zoology, Kunming, 650223, People’s Republic of China

      Bingyu Mao

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    1. Shuying Yu
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    2. Xuhui Chen
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    3. Zuoqing Yuan
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    4. Luming Zhou
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    7. Bosheng Zhao
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    Corresponding author

    Correspondence to Bosheng Zhao.

    Additional information

    Communicated by S. Hohmann.

    S. Yu and X. Chen contributed equally to this work.

    GenBank Accession Number: HQ585080.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    The nucleotide and deduced amino acid sequences of DjElc cDNA. EF-Hand is boxed (TIFF 4407 kb)

    438_2015_990_MOESM2_ESM.tif

    Alignment of ELCs, including DjELC using the MegAlign program (DNASTAR) by the CLUSTAL W method. Shaded (with solid black) residues are the amino acids that match the consensus. Gaps introduced into sequences to optimize alignment are represented by (−). “#” means Ca2+ binding sites. ELC amino acid sequences were obtained from NCBI: Apis mellifera (XP_393544); Ascaris suum (ERG85616); Branchiostoma floridae (XP_002603233.1); Bos taurus (NP_001071124); Caenorhabditis brenneri (ACD86909); Caenorhabditis elegans (P53014); Dictyostelium discoideum (P09402); Danio rerio (NP_001005955); Dugesia japonica (HQ585080); Drosophila melanogaster (AAF56733); Echinococcus granulosus (CDJ16649); Gallus gallus (NP_001038097); Hymenolepis microstoma (CDJ14659); Homo sapiens (NP_524144); Marsupenaeus japonicas (ADD70028); Mus musculus (NP_067260); Pan troglodytes (XP_516064); Rattus norvegicus (NP_001071124); Setaria digitata (ACT15365); Schistosoma japonicum (CAX70946); Schistosoma mansoni (AAD41591); Xenopus tropicalis (NP_001121402) (TIFF 10759 kb)

    438_2015_990_MOESM3_ESM.tif

    Expression level of DjElc mRNA in regenerating trunk fragments of DjElc-RNAi-treated planarians by quantitative real-time PCR (TIFF 699 kb)

    438_2015_990_MOESM4_ESM.tif

    Ventral view of defects of DjElc-RNAi-treated trunk fragments to regenerate head by immunostaining with anti-synaptotagmin. (A) Normal regenerating head at 7 days as a control. (B) Loss of DjElc disrupted the brain lateral branches at 7 days. Scale bars: 50 μm (TIFF 1329 kb)

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    Yu, S., Chen, X., Yuan, Z. et al. Planarian myosin essential light chain is involved in the formation of brain lateral branches during regeneration. Mol Genet Genomics 290, 1277–1285 (2015). https://doi.org/10.1007/s00438-015-0990-1

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    • DOI: https://doi.org/10.1007/s00438-015-0990-1

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