Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of pigment cells in the tunicate Ciona intestinalis to study pigment cell specification within Central Nervous System

ABSTRACT: Its known that FGF signaling induces pigment cell precursors formation. We perturbed the endogenous FGF signaling cascade by using a dominant-negative form of the unique Ci-FGF receptor (FGFRDN) and a constitutively active form of Ci-Ets1/2 (Ets:Vp16), a transcriptional effector of FGF/MAP Kinase cascade. Targeted expression of dominant negative FGF receptor and Ets:VP16 fusion protein was achieved in PCPs using the Tyrosinase-related protein 1/2a (Tyrp1/2a) cis-regulatory region. The PCPs were isolated by Fluorescent Activated Cell Sorting (FACS) as GFP+ cells at two developmental stages: 8 hpf at ~16C (neurula stage) corresponding to early FGF-mediated induction and 12 hpf at ~18C (tailbud stage) where PCPs are already fate restricted as pigment cells. To avoid a contamination by mesenchyme cells where Ciona enhancers are often ectopically activated, we co-electroporated the MyoD905>YFP construct and counter-selected YFP+ cells. In summary the conditions used are: FGFRDN, Ets:VP16 and Control samples at 8 and 12 hpf (four samples for each condition).

ORGANISM(S): Ciona intestinalis

SUBMITTER: Claudia Racioppi

PROVIDER: E-MTAB-2759 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Eight different types of sample (three samples each) were used in this study of cardiac cell migration in the tunicate Ciona intestinalis. The B7.5 lineage cells were isolated by Fluorescent Activated Cell Sorting (FACS) at the tailbud stage, when the cardiac precursors (anterior B7.5 lineage cells) are migrating into the trunk, while their sister cells (posterior 7.5 lineage cells) remain in the tail, where they form muscle cells. Cells were sorted at the late gastrula stage, prior to migration induction and after manipulation of the FGF-MAPK-Ets signaling pathway (targeted expression of dominant negative FGF receptor and Ets:VP16 fusion protein, targeted expression was achieved using the Mesp cis-regulatory DNA), FoxF function (a direct target of FGF-MAPK-Ets signaling, required primarily for cell migration) by targeted expression of FoxF:VP16 (constitutive activator) or FoxF:WRPW (constitutive repressor, dominant negative), Mesp function (over-expression of Mesp:VP16 inhibits primarily cell migration). Finally, the "whole embryo" samples were collected from dissociated whole tailbud embryos, without FACS. in summary the eight conditions are: "wt (files LC-wt-1,-2,-3)": wild_type B7.5 lineage cells, expressing GFP, sorted at the tailbud stage "Ets:VP16 (files LC-EV-1,-2,-3)": B7.5 lineage cells, expressing GFP and Ets:VP16, sorted at the tailbud stage "DnFGFR (files LC-dnFGFR-1,-2,-3)": B7.5 lineage cells, expressing GFP and dominant-negative FGF receptor, sorted at the tailbud stage. "FoxF:VP16 (files LC-FV-1,-2,-3)": B7.5 lineage cells, expressing GFP and FoxF:VP16, sorted at the tailbud stage. "FoxF:WRPW (files LC-FW-1,-2,-3)": B7.5 lineage cells, expressing GFP and FoxF:WRPW, sorted at the tailbud stage". "Mesp:VP16 (files LC-MV-1,-2,-3)": B7.5 lineage cells, expressing GFP and Mesp:VP16, sorted at the tailbud stage. "Late Gastrula (files LC-LG-1,-2,-3)": wild_type B7.5 lineage cells, expressing GFP, sorted at the late gastrula stage "whole embryo (files LC-whole-1,-2,-3": whole embryos, dissociated at the tailbud stage, not sorted.

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Transcription profiling of pigment cells in the tunicate Ciona intestinalis to study pigment cell specification within Central Nervous System

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