Open Access Highly Accessed Methodology article

Development of a microinjection system for RNA interference in the water flea Daphnia pulex

Chizue Hiruta1, Kenji Toyota12, Hitoshi Miyakawa1, Yukiko Ogino12, Shinichi Miyagawa12, Norihisa Tatarazako3, Joseph R Shaw4 and Taisen Iguchi12*

Author Affiliations

1 Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan

2 Department of Basic Biology, Faculty of Life Science, Graduate University for Advanced Studies (SOKENDAI), 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan

3 Environmental Quality Measurement Section, Research Center for Environmental Risk, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506, Japan

4 School of Public and Environmental Affairs, Indiana University, 1315 East Tenth Street, Bloomington IN 47405, USA

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BMC Biotechnology 2013, 13:96  doi:10.1186/1472-6750-13-96

Published: 5 November 2013

Additional files

Additional file 1:

Characteristics of egg in D. pulex (A, C) and D. magna (B, D). Samples were collected just after ovulation. (A, B) Light micrographs. (C, D) Hematoxylin and eosin (HE)-stained cross section. od, oil droplet; yg, yolk granule. Scale bar = 100 μm.

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Additional file 2:

Nucleotide sequences of Dll-dsRNA.

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Additional file 3:

Phenotypes of Dll-dsRNA injected juveniles. The left and right columns show phenotypes of individuals injected with dsRNA of malE and Dll, respectively. (A, B) First thoracic limb (T1). The exopodite and endopodite were shortened by Dll-dsRNA. (C, D) Third and fourth thoracic limbs (T3/4), having the same morphology. The exopodite was shrunk in Dll-dsRNA-injected juveniles. (E, F) Fifth thoracic limb (T5). The exopodite was shortened and twisted by Dll-dsRNA. (G, H) Lateral view of the rostrum and head. An arrowhead indicates an ocellus. (I, J) Lateral view of abdomens. An arrow and arrowhead show an abdominal claw and abdominal setae. Ep, epipodite; En, endopodite; Ex, exopodite; Fc, filter comb. Scale bars = 100 μm.

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Additional file 4:

Primer sequences for in situ hybridization, dsRNA and Q-PCR.

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