The Zebrafish: Disease Models and Chemical Screens, Volume 105

Contributors

Chapter 1: Zebrafish Models of Germ Cell Tumor

I. Overview

II. Germline Development

III. Germ Cell Tumors

IV. Methods for Studying Zebrafish Germ Cells

Chapter 2: Dissecting Mechanisms of Myelinated Axon Formation Using Zebrafish

I. Introduction

II. Methods and Materials

III. Summary

Chapter 3: In vivo Analysis of White Adipose Tissue in Zebrafish

I. Introduction

II. Rationale

III. Materials

IV. Methods

V. Summary

Chapter 4: Study of Host–Microbe Interactions in Zebrafish

I. Introduction

II. Laboratory Protocols

Chapter 5: Hematopoietic Stem Cell Development: Using the Zebrafish to Identify the Signaling Networks and Physical Forces Regulating Hematopoiesis

I. Developmental Hematopoiesis

II. Review of the Literature

III. Use of Zebrafish to Investigate HSC Biology

IV. Zebrafish Tools and Protocols

Chapter 6: Zebrafish as a Model for Hemorrhagic Stroke

I. Introduction and Basic Concepts

II. Maintaining the Barrier Function of Blood Vessels

III. Vascular Integrity and Stroke

IV. Zebrafish as a Model for Studying Hemorrhage and Stroke

V. Studying Vascular Integrity in the Zebrafish

VI. Concluding Remarks

Chapter 7: A Zebrafish Model for VHL and Hypoxia Signaling

I. VHL Disease

II. Chuvash Polycythemia

III. VHL Regulates HIF Signaling

IV. HIF-Independent Functions of pVHL

V. VHL Animal Models

VI. Zebrafish as a New Model for VHL

VII. Loss of Vhl Leads to a Systemic Hypoxic Response in Zebrafish

VIII. vhl Mutants Develop Chuvash Polycythemia and Blood Cell Maturation Defects

IX. Zebrafish vhl Mutants Develop Angiogenesis Defects

X. Zebrafish vhl Mutants Develop Pronephros Abnormalities

XI. Translation from Fish to Humans

XII. Concluding Remarks

Chapter 8: Basement Membrane Diseases in Zebrafish

I. Introduction

II. Basement Membrane-Related Zebrafish Pathologies

III. Methodology for Zebrafish Studies of Basement Membrane Composition and Function

IV. Summary

Chapter 9: Zebrafish Provides a Novel Model for Lymphatic Vascular Research

I. Introduction

II. Embryonic Lymphangiogenesis in Zebrafish

III. Comparison Between Teleosts and Other Vertebrates

IV. The Added Value of Zebrafish

V. The Secondary Vessel System in Teleosts

VI. Concluding Remarks

VII. Methods

Chapter 10: Not All Bones are Created Equal – Using Zebrafish and Other Teleost Species in Osteogenesis Research

I. Case Studies – Using Zebrafish for Addressing Biomedical Questions

II. The Evolution of Skeletal Tissues

III. Cartilage and Bone in Teleost Fish

IV. Intermediate Skeletal Tissues

V. Osteocyte-Containing Bone and Acellular Bone

VI. Development of Teleost Vertebral Bodies, A Derived Process

VII. Remodeling of the Teleost Skeleton

VIII. Conclusions

Chapter 11: Zebrafish Assays of Ciliopathies

I. Introduction

II. Methods

III. Zebrafish Ciliary Mutant Lines

IV. General Considerations/Future Development

Chapter 12: Infectious Disease Modeling and Innate Immune Function in Zebrafish Embryos

I. Introduction

II. Observation and Isolation of Innate Immune Cells

III. Bacterial Infection Methods

IV. Analysis of the Innate Immune Response

V. Conclusions

Chapter 13: Zebrafish as a Model for the Study of Human Cancer

I. Introduction: Zebrafish as a Cancer Model

II. The Genetically Tractable Zebrafish

III. Transgenic Models of Oncogenesis

IV. Modeling the Loss of Tumor Suppression

V. Modeling Tumor Cell Intravasation and Metastasis

VI. Tumor-Initiating Cells

VII. In vivo Small-Molecule Screens and Drug Discovery

VIII. Conclusions

Chapter 14: Generating and Analyzing Fish Models of Melanoma

I. Introduction

II. Transgenic Melanoma Lines in Zebrafish and Medaka

III. Ultra-violet Irradiation Treatments

IV. Basic Melanoma Pathology

V. Histology

VI. Molecular Analysis

VII. RNA and Protein Isolation From Adult Fish Tumors

VIII. Conclusions

Chapter 15: Screening Pancreatic Oncogenes in Zebrafish Using the Gal4/UAS System

I. Introduction

II. Transgenic Zebrafish with Gal4/UAS-Mediated eGFP-KRASG12V Expression in the Exocrine Pancreas

III. Identification and Characterization of Pancreatic Tumors

IV. Conclusions

Chapter 16: Zebrafish Models of Rhabdomyosarcoma

I. Introduction

II. Rationale

III. Material and Methods

IV. Discussion

Chapter 17: Transplantation in Zebrafish

I. Introduction

II. Rationale

III. Methods

IV. Discussion

Chapter 18: Disease Modeling by Gene Targeting Using MicroRNAs

I. Introduction

II. Mechanisms of MicroRNA Silencing

III. Dicer and MicroRNAs in Zebrafish Development

IV. Development of Vector-Based RNA Interference

V. RNA Interference Work in Zebrafish

VI. Use of siRNAs in the Zebrafish

VII. Materials and Methods

VIII. Results

IX. Future Directions

Chapter 19: Fluorescent Imaging of Cancer in Zebrafish

I. Introduction

II. Fluorescent Proteins and Transgenic Models of Cancer

III. Macroscopic Observation of Tumor Growth

IV. Microscopic Observation in Tumorigenesis

V. Confirming Transformation of Fluorescent-Labeled Tumor Cells by Cell Transplantation into Irradiated Recipient Animals

VI. Identifying Tumor-Propagating Cell Subpopulations by Fluorescent Protein Expression and Cell Transplantation into Irradiated Recipient Animals

VII. Use of Syngeneic Zebrafish for Cell Transplantation of Fluorescent-Labeled Tumors and Drug Discovery

VIII. Cell Transplantation into Syngeneic Zebrafish to Accurately Assess Self-Renewal in Fluorescent-Labeled Cancer

IX. Use of Syngeneic Zebrafish for Cell Transplantation: Single Cell Transplants

X. Xenograft Transplantation of Fluorescently Labeled Cells into Zebrafish

XI. Conclusions

Chapter 20: The Role of Fanconi Anemia/BRCA Genes in Zebrafish Sex Determination

I. Introduction

II. Results and Discussion

III. Summary

Chapter 21: Chemical Screening in Zebrafish for Novel Biological and Therapeutic Discovery

I. Introduction

II. Rationale

III. Materials and Methods

IV. Discussion/Caveats

V. Summary

Chapter 22: Using the Zebrafish Photomotor Response for Psychotropic Drug Screening

I. Introduction

II. The PMR Behavior

III. Methods

IV. Discussion

Chapter 23: Designing Zebrafish Chemical Screens

I. Rationale for Conducting Small Molecule Screens With Zebrafish

II. Selection of Small Molecule Libraries

III. Assay Design: Major Screen Types

IV. Screening Methods

V. Mechanism of Action Studies

VI. Conclusions

Preface

Index

Volumes in Series

This volume of Methods in Cell Biology is the 3e, and provides comprehensive compendia of laboratory protocols and reviews covering all the new methods developed since 2004. This new volume on Disease Models and Chemical Screens, covers two rapidly emerging and compelling applications of the zebrafish.

• Details state-of-the art zebrafish protocols, delineating critical steps in the procedures as well as potential pitfalls
• This volume concentrates on Disease Models and Chemical Screens

Developmental biologists, neurobiologists, and cell biologists