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Research Techniques Made Simple: Zebrafish as a Model System to Study Skin Biology and Pathology

Research Techniques Made Simple: Zebrafish as a Model System to Study Skin Biology and Pathology. Qiaoli Li and Jouni Uitto

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Research Techniques Made Simple: Zebrafish as a Model System to Study Skin Biology and Pathology

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  1. Research Techniques Made Simple:Zebrafish as a Model System to Study Skin Biology and Pathology Qiaoli Li and JouniUitto Department of Dermatology and Cutaneous Biology, Jefferson Medical College, and Jefferson Institute of Molecular Medicine, Thomas Jefferson University, Philadelphia, PA, USA

  2. Zebrafish and Embryos • Vertebrate • Has nearly the same complement of genes as mammals • Develop externally • Transparent • Easy breeding • Develop quickly

  3. Zebrafish Skin Development Reprinted with permission from Li et al. (2011) J Invest Dermatol 131:565-71

  4. Genetic Manipulation of Zebrafish Genome • Forward genetics Ethylnitrosourea (ENU) or random mutagenesis • Reverse genetics Morpholino knockdown of specific gene expression

  5. Modified from (Dahm, 2005) Encyclopedia of Molecular Cell Biology and Molecular Medicine (Meyers RA, ed) Vol. 15: Wiley-VCH, 593-626, with permission. ENU Mutagenesis

  6. Modulation of Gene Expression by Morpholino 25 nt oligo “Knockdown”

  7. Efficiency of Morpholino Knockdown ATG morpholino Splice morpholino ATG abcc6bMO2 MO1 5’UTR forward target gene EXON 19 EXON 18 reverse EGFP SP6 promoter pCS2/EGFP vector MO2 C 500bp 421bp col17a1b/EGFP RNA col17a1b/EGFP RNA + col17a1b MO1 299bp 300bp 8.5hpf 8.5hpf 8.5hpf Reprinted with permission from Li et al. (2011) J Invest Dermatol 131:565-71

  8. Application of Zebrafish for Skin Research • Wound healing and skin repair • Pigmentation • Epidermal adhesion • Skin diseases • Skin immunity • Drug screening

  9. Advantages • A freshwater vertebrate with a complement of genes very similar to those of mammals • Rapid development of transparent embryos, allowing visual inspection of internal organs • Easy breeding, with a large number of embryos and larvae • Easy manipulation of gene expression with phenotypic consequences • A facile and cost-efficient system for large-scale screening

  10. Limitations • Lack of functional stratum corneum and expression of genes necessary for terminal differentiation of epidermis • The zebrafish genome database is not complete and contains misjoins and misassemblies • Paucity of antibodies recognizing zebrafish protein sequences

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