Supplementary MaterialsFigure 1source data 1: Hair cell survival post neomycin in crazy type and larvae. crazy type and hair cells. elife-47061-fig5-data3.xlsx (13K) DOI:?10.7554/eLife.47061.021 Number 5source data 4: Hair cell survival post Antimycin A in wild type and larvae. elife-47061-fig5-data4.xlsx (16K) DOI:?10.7554/eLife.47061.022 Number 5figure product 1source data 1: Mean F(mitotracker) in wild type and hair cells. elife-47061-fig5-figsupp1-data1.xlsx (11K) DOI:?10.7554/eLife.47061.017 Number 5figure product 1source data 2: Mean F(CellROX) and percentage of mean F(CellROX) to mean F(GFP) in wild type and pappaa mutant hair cells. elife-47061-fig5-figsupp1-data2.xlsx (12K) DOI:?10.7554/eLife.47061.018 Figure 6source data 1: Mean Aprotinin F(mitoGCaMP) in wild type and hair cells. elife-47061-fig6-data1.xlsx (12K) DOI:?10.7554/eLife.47061.024 Number 6source data 2: Mean F(TMRE) and percentage of mean F(TMRE) to F(GFP) in wild type and hair cells. elife-47061-fig6-data2.xlsx (14K) DOI:?10.7554/eLife.47061.025 Number 6source data 3: Hair cell survival post Cyclosporin A in wild Aprotinin type and larvae. elife-47061-fig6-data3.xlsx (18K) DOI:?10.7554/eLife.47061.026 Number 7source data 1: Quantification of antioxidant transcript expression in wild type and hair cells. elife-47061-fig7-data1.xlsx (17K) DOI:?10.7554/eLife.47061.028 Figure 7source data 2: Hair cell survival post co-treatment of mitoTEMPO and neomycin in larvae. elife-47061-fig7-data2.xlsx (17K) DOI:?10.7554/eLife.47061.029 Transparent reporting form. elife-47061-transrepform.docx (250K) DOI:?10.7554/eLife.47061.030 Data Availability StatementAll data generated or analysed during this study are included in the manuscript and assisting files. Source data files have been offered for Numbers 1,3,4,5,6,7 and all supplementary numbers. Abstract To support cell survival, mitochondria must balance energy production with oxidative stress. Inner hearing hair cells are particularly vulnerable to oxidative stress; therefore require limited mitochondrial rules. We recognized a novel molecular regulator of the hair cells mitochondria and survival: Pregnancy-associated plasma protein-aa (Pappaa). Hair Rabbit Polyclonal to NT cells in zebrafish mutants show mitochondrial defects, including elevated mitochondrial calcium, transmembrane potential, and reactive oxygen species (ROS) production and reduced antioxidant manifestation. In mutants, hair cell death is definitely enhanced by activation of mitochondrial calcium or ROS production and suppressed by a mitochondrial ROS scavenger. Like a secreted metalloprotease, Pappaa stimulates extracellular insulin-like growth element 1 (IGF1) bioavailability. We found that the mutants enhanced hair cell loss can be suppressed by activation of IGF1 availability and that Pappaa-IGF1 signaling functions post-developmentally to support hair cell survival. These results reveal Pappaa as an extracellular regulator of hair cell survival and essential mitochondrial function. mutants, we reveal a novel part for Pappaa in regulating mitochondrial function to support hair cell survival. Results IGF1R signaling affects hair cell survival and mitochondrial Aprotinin function in zebrafish Hair cells of the zebrafish lateral collection are found in superficial neuromasts and form a rosette-like structure that is surrounded by support cells (Raible and Kruse, 2000)?(Number 1A). These hair cells share practical, morphological, and molecular similarities with mammalian inner ear hair cells (Ghysen and Dambly-Chaudire, 2007). Acute exposure of larval zebrafish to the aminoglycoside neomycin causes hair cell death and mitochondrial dysfunction (Harris et al., 2003; Esterberg et al., 2014; Esterberg et al., 2016). This experimental platform has been used to dissect the molecular and cellular mechanisms that support hair cell survival (Owens et al., 2008). A role for IGF1R signaling in the survival of zebrafish lateral collection hair cells and their mitochondria offers yet to be shown. We hypothesized that if IGF1R signaling helps hair cell survival, then attenuating IGF1R signaling would further reduce hair cell survival following neomycin exposure. To test this, we used a transgenic collection in which an inducible warmth shock promoter drives ubiquitous manifestation of a dominating bad IGF1Ra [(manifestation was induced from 24 hr post fertilization (hpf) to 5 days post fertilization (dpf). At five dpf, larvae were exposed to neomycin for 1 hr and evaluated for hair cell survival 4 hr later on. Larvae expressing showed a greater reduction in.
