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Stable knock-down of vomeronasal receptor genes in transgenic Xenopus tadpoles
Authors:Kashiwagi Akihiko  Kashiwagi Keiko  Saito Shouichiro  Date-Ito Atsuko  Ichikawa Masumi  Mori Yuji  Hagino-Yamagishi Kimiko
Institution:Institute for Amphibian Biology, Graduate School of Science, Hiroshima University, 1-3-1, Kagamiyama, Higashihiroshima 739-8526, Japan. akashiwa@hiroshima-u.ac.jp
Abstract:Xenopus V2R (xV2R), a family of G-protein-coupled receptors with seven transmembrane domains, is expressed in the Xenopus vomeronasal organ (VNO). There are six subgroups of xV2R, one of which, xV2RE, is predominantly expressed in the VNO. To understand the function of xV2R during VNO development, we developed a new method to achieve stable siRNA-suppression of the V2RE genes by introducing siRNA expression transgenes into the genomes of unfertilized eggs. We found that some of the derived transgenic tadpoles lacked VNOs and that their olfactory epithelium was fused. With the exception of one tadpole, expression of xV2RE was not detected in morphologically abnormal mutant tadpoles, although the olfactory marker protein and the olfactory receptors were expressed. These results suggest that we successfully produced transgenic tadpoles in which xV2RE expression was stably suppressed by siRNA, and that xV2RE plays a role in the morphogenesis of olfactory organs.
Keywords:siRNA  V2R  VNO  Olfactory epithelium  Xenopus laevis  Transgenic
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