Xenbase Image ID: 138197

Fig. 4. A. Intracellular Ca2+ signaling in the kidney field requires TRPP2. (A) Schematic illustrating the procedure for Ca2+ measurements in the kidney field. CMO or Pkd2-MO morpholinos were injected in the V2 blastomere of eight-cell stage embryos along with GFP–aequorin mRNA. Kidney field (KF) explants were dissected at the late gastrula stage (NF stage 12.5) and Ca2+ transients were measured with a PMT for 4 h, until sibling developing embryos reach NF stage 18 (late neurula). (B) PMT traces from single pronephric territories dissected from control morpholino-injected embryo (CMO) and TRPP2 morpholino-injected embryo (Pkd2-MO). Data is representative of pairs of territories measured simultaneously in four independent experiments. (C) Histogram plot displaying the mean6s.e.m. number of Ca2+ transients during 4 h. The two left bars correspond to the mean6s.e.m. number of transients calculated for pairs of CMO and Pkd2-MO explants respectively (n54 pairs). In this condition, Pkd2 knockdown significantly reduces the number of Ca2+ transients (P50.014, paired Student’s ttest). The two right bar plots correspond to pairs of Pkd2-MO and rescue (Pkd2-MO + pkd2 mRNA) explants, respectively (n53 pairs). Although the number of Ca2+ transients increases in the rescue condition, it is not significant (see results for details). (D) Rescue of Ca2+ transients by pkd2 expression in Pkd2-MO explants. Pkd2-MO and GFP–aequorin mRNA were injected alone or mixed with pkd2 mRNA. PMT traces from single pronephric territories dissected from Pkd2- MO-injected embryo (Pkd2-MO) and embryo co-injected with Pkd2-MO and a Pkd2-MO resistant form of pkd2 mRNA are shown. Data is representative of pairs of territories measured simultaneously from three independent experiments. Image published in: Futel M et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, The Company of Biologists Ltd. Larger Image Printer Friendly View

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