Transgenesis in the acoel worm Hofstenia miamia

The acoel worm Hofstenia miamia, which can replace tissue lost to injury via differentiation of a population of stem cells, has emerged as a new research organism for studying regeneration. To enhance the depth of mechanistic studies in this system, we devised a protocol for microinjection into embryonic cells that resulted in stable transgene integration into the genome and generated animals with tissue-specific fluorescent transgene expression in epidermis, gut, and muscle. We demonstrate that transgenic Hofstenia are amenable to the isolation of specific cell types, detailed investigations of regeneration, tracking of photoconverted molecules, and live imaging. Further, our stable transgenic lines revealed new insights into the biology of Hofstenia, unprecedented details of cell morphology and the organization of muscle as a cellular scaffold for other tissues. Our work positions Hofstenia as a powerful system with unparalleled tools for mechanistic investigations of development, whole-body regeneration, and stem cell biology.

[1]  A. De Tomaso,et al.  Integrin-alpha-6+ Candidate stem cells are responsible for whole body regeneration in the invertebrate chordate Botrylloides diegensis , 2020, Nature Communications.

[2]  Ryan E. Hulett,et al.  Neural architecture and regeneration in the acoel Hofstenia miamia , 2020, Proceedings of the Royal Society B.

[3]  P. Reddien,et al.  Muscle and neuronal guidepost-like cells facilitate planarian visual system regeneration , 2020, Science.

[4]  M. Srivastava,et al.  A regulatory program for initiation of Wnt signaling during posterior regeneration , 2020, bioRxiv.

[5]  U. Jondelius,et al.  Xenacoelomorpha, a Key Group to Understand Bilaterian Evolution: Morphological and Molecular Perspectives , 2019, Evolution, Origin of Life, Concepts and Methods.

[6]  P. Reddien,et al.  A small set of conserved genes, including sp5 and Hox, are activated by Wnt signaling in the posterior of planarians and acoels , 2019, PLoS genetics.

[7]  V. Hartenstein,et al.  The digestive system of xenacoelomorphs , 2019, Cell and Tissue Research.

[8]  P. Reddien,et al.  Muscle functions as a connective tissue and source of extracellular matrix in planarians , 2019, Nature Communications.

[9]  Ryan E. Hulett,et al.  Acoel genome reveals the regulatory landscape of whole-body regeneration , 2019, Science.

[10]  Jens H. Fritzenwanker,et al.  I-SceI Meganuclease-mediated transgenesis in the acorn worm, Saccoglossus kowalevskii. , 2019, Developmental biology.

[11]  B. Goldstein.  The Emergence of the Tardigrade Hypsibius exemplaris as a Model System. , 2018, Cold Spring Harbor protocols.

[12]  A. Brunet,et al.  The African turquoise killifish: A research organism to study vertebrate aging and diapause , 2018, Aging cell.

[13]  E. Inada,et al.  Timing of CRISPR/Cas9-related mRNA microinjection after activation as an important factor affecting genome editing efficiency in porcine oocytes. , 2018, Theriogenology.

[14]  Irving E. Wang,et al.  Clonal Analysis of Planarian Stem Cells by Subtotal Irradiation and Single-Cell Transplantation. , 2018, Methods in molecular biology.

[15]  Eugene Berezikov,et al.  Efficient transgenesis and annotated genome sequence of the regenerative flatworm model Macrostomum lignano , 2017, Nature Communications.

[16]  P. Reddien,et al.  Orthogonal muscle fibers have different instructive roles in planarian regeneration , 2017, Nature.

[17]  P. Reddien,et al.  Acoel regeneration mechanisms indicate an ancient role for muscle in regenerative patterning , 2017, Nature Communications.

[18]  James M. Gahan,et al.  The interstitial stem cells in Hydractinia and their role in regeneration. , 2016, Current opinion in genetics & development.

[19]  R. D. Reed,et al.  Genome editing in butterflies reveals that spalt promotes and Distal-less represses eyespot colour patterns , 2016, Nature Communications.

[20]  F. Ronquist,et al.  Xenacoelomorpha is the sister group to Nephrozoa , 2016, Nature.

[21]  S. McKinney,et al.  TALEN and CRISPR/Cas9-mediated genome editing in the early-branching metazoan Nematostella vectensis , 2014, Nature Communications.

[22]  P. Reddien,et al.  Whole-Body Acoel Regeneration Is Controlled by Wnt and Bmp-Admp Signaling , 2014, Current Biology.

[23]  P. Reddien,et al.  Muscle Cells Provide Instructions for Planarian Regeneration , 2013, Cell reports.

[24]  F. Raible,et al.  Stable transgenesis in the marine annelid Platynereis dumerilii sheds new light on photoreceptor evolution , 2012, Proceedings of the National Academy of Sciences.

[25]  Ulrich C. Klostermeier,et al.  Molecular signatures of the three stem cell lineages in hydra and the emergence of stem cell function at the base of multicellularity. , 2012, Molecular biology and evolution.

[26]  P. Martinez,et al.  The Acoela: on their kind and kinships, especially with nemertodermatids and xenoturbellids (Bilateria incertae sedis) , 2012, Organisms Diversity & Evolution.

[27]  Matthew S. Burriesci,et al.  Developing the anemone Aiptasia as a tractable model for cnidarian-dinoflagellate symbiosis: the transcriptome of aposymbiotic A. pallida , 2012, BMC Genomics.

[28]  Patrick R. H. Steinmetz,et al.  A muscle-specific transgenic reporter line of the sea anemone, Nematostella vectensis , 2009, Proceedings of the National Academy of Sciences.

[29]  C. Todt.  Structure and evolution of the pharynx simplex in acoel flatworms (Acoela) , 2009, Journal of morphology.

[30]  C. Todt,et al.  Ciliary receptors associated with the mouth and pharynx of Acoela (Acoelomorpha): a comparative ultrastructural study , 2006 .

[31]  H. Ogino,et al.  Highly efficient transgenesis in Xenopus tropicalis using I-SceI meganuclease , 2006, Mechanisms of Development.

[32]  Julian P. S. Smith.  Fine-structural observations on the central parenchyma in Convoluta sp. , 2004, Hydrobiologia.

[33]  William C. Smith,et al.  Generation of Ci‐Brachyury‐GFP stable transgenic lines in the ascidian Ciona savignyi , 2003, Genesis.

[34]  J. Joly,et al.  I-SceI meganuclease mediates highly efficient transgenesis in fish , 2002, Mechanisms of Development.

[35]  S. Tyler,et al.  Functional morphology of musculature in the acoelomate worm, Convoluta pulchra (Plathelminthes) , 1999, Zoomorphology.