Characterizing the spatiotemporal expression of rnas and proteins in the starlet sea anemone, nematostella vectensis

ABSTRACT In an effort to reconstruct the early evolution of animal genes and proteins, there is an increasing focus on basal animal lineages such as sponges, cnidarians, ctenophores and

placozoans. Among the basal animals, the starlet sea anemone _Nematostella vectensis_ (phylum Cnidaria) has emerged as a leading laboratory model organism partly because it is well suited to

experimental techniques for monitoring and manipulating gene expression. Here we describe protocols adapted for use in _Nematostella_ to characterize the expression of RNAs by _in situ_

hybridization using either chromogenic or fluorescence immunohistochemistry (∼1 week), as well as to characterize protein expression by whole-mount immunofluorescence (∼3 d). We also provide

a protocol for labeling cnidocytes (∼3 h), the phylum-specific sensory-effector cell type that performs a variety of functions in cnidarians, including the delivery of their venomous sting.

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ACKNOWLEDGEMENTS This research was supported by National Science Foundation grant no. MCB-0924749 to T.D.G. and J.R.F. and by the US National Institutes of Health (NIH) grant no.

1R21RR032121 to M.Q.M. F.S.W. was supported by a predoctoral grant from the Superfund Basic Research Program at Boston University (no. 5 P42 E507381) and by Warren-McLeod graduate

fellowships in Marine Biology. M.J.L. was supported by a Ruth L. Kirschstein National Research Service Award (no. FHD0550002) from the NIH. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS *

Department of Biology, Boston University, Boston, Massachusetts, USA Francis S Wolenski, Thomas D Gilmore & John R Finnerty * Pacific Biosciences Research Center, Kewalo Marine

Laboratory, University of Hawaii, Honolulu, Hawaii, USA Michael J Layden & Mark Q Martindale Authors * Francis S Wolenski View author publications You can also search for this author

inPubMed Google Scholar * Michael J Layden View author publications You can also search for this author inPubMed Google Scholar * Mark Q Martindale View author publications You can also

search for this author inPubMed Google Scholar * Thomas D Gilmore View author publications You can also search for this author inPubMed Google Scholar * John R Finnerty View author

publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS F.S.W. optimized immunofluorescence and cnidocyte-labeling protocols. M.J.L. optimized the _in situ_

hybridization protocols. J.R.F., T.D.G. and M.Q.M. provided technical advice on protocol development. All authors participated in writing the manuscript. CORRESPONDING AUTHOR Correspondence

to John R Finnerty. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS

ARTICLE Wolenski, F., Layden, M., Martindale, M. _et al._ Characterizing the spatiotemporal expression of RNAs and proteins in the starlet sea anemone, _Nematostella vectensis_. _Nat

Protoc_ 8, 900–915 (2013). https://doi.org/10.1038/nprot.2013.014 Download citation * Published: 11 April 2013 * Issue Date: May 2013 * DOI: https://doi.org/10.1038/nprot.2013.014 SHARE THIS

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