title: Manipulation of Interrenal Cell Function in Developing Zebrafish Using Genetically Targeted Ablation and an Optogenetic Tool.
abstract: Zebrafish offer an opportunity to study conserved mechanisms underlying the ontogeny and physiology of the hypothalamic-pituitary-adrenal/interrenal axis. As the final effector of the hypothalamic-pituitary-adrenal/interrenal axis, glucocorticoids exert both rapid and long-term regulatory functions. To elucidate their specific effects in zebrafish, transgenic approaches are necessary to complement pharmacological studies. Here, we report a robust approach to specifically manipulate endogenous concentrations of cortisol by targeting heterologous proteins to interrenal cells using a promoter element of the steroidogenic acute regulatory protein. To test this approach, we first used this regulatory region to generate a transgenic line expressing the bacterial nitroreductase protein, which allows conditional targeted ablation of interrenal cells. We demonstrate that this line can be used to specifically ablate interrenal cells, drastically reducing both basal and stress-induced cortisol concentrations. Next, we coupled this regulatory region to an optogenetic actuator, Beggiatoa photoactivated adenylyl cyclase, to increase endogenous cortisol concentrations in a blue light-dependent manner. Thus, our approach allows specific manipulations of steroidogenic interrenal cell activity for studying the effects of both hypo- and hypercortisolemia in zebrafish.
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Jose Arturo
authors.initials: JA
authors.lastname: Gutierrez-Triana
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Herget
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Luis A
authors.initials: LA
authors.lastname: Castillo-Ramirez
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Lutz
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Chen-Min
authors.initials: CM
authors.lastname: Yeh
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Rodrigo J
authors.initials: RJ
authors.lastname: De Marco
authors.affiliation: Developmental Genetics of the Nervous System (J.A.G.-T., U.H., L.A.C.-R., M.L., C.-M.Y., R.J.D.M., S.R.), Max Planck Institute for Medical Research, D-69120 Heidelberg, Germany; and The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (U.H., L.A.C.-R.), University of Heidelberg, D-69120 Germany.
authors.forename: Soojin
authors.initials: S
authors.lastname: Ryu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D006854:Hydrocortisone / Q000378:metabolism*; D007401:Interrenal Gland / Q000378:metabolism*; D062308:Optogenetics; D010750:Phosphoproteins / Q000235:genetics*; D050436:Regulatory Elements, Transcriptional; D015027:Zebrafish; D000098682:Steroidogenic Acute Regulatory Protein
chemical_list: D010750:Phosphoproteins; D000098682:Steroidogenic Acute Regulatory Protein; D006854:Hydrocortisone