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J Vis Exp. 2016 Jun 11;(112). doi: 10.3791/54168.

Generation of Parabiotic Zebrafish Embryos by Surgical Fusion of Developing Blastulae.

Author information

1
Division of Hematology/Oncology, Boston Children's Hospital; Harvard Medical School.
2
Division of Hematology, Department of Medicine, Brigham and Women's Hospital.
3
Division of Hematology/Oncology, Boston Children's Hospital; Harvard Medical School; Division of Hematologic Malignancies, Dana-Farber Cancer Institute.
4
Division of Hematology/Oncology, Boston Children's Hospital; Harvard Medical School; Harvard Stem Cell Institute; Broad Institute of Massachusetts Institute of Technology; Howard Hughes Medical Institute.
5
Division of Hematology/Oncology, Boston Children's Hospital; Harvard Medical School; Division of Hematology, Department of Medicine, Brigham and Women's Hospital; Harvard Stem Cell Institute; Broad Institute of Massachusetts Institute of Technology; DSHAH@research.bwh.harvard.edu.

Abstract

Surgical parabiosis of two animals of different genetic backgrounds creates a unique scenario to study cell-intrinsic versus cell-extrinsic roles for candidate genes of interest, migratory behaviors of cells, and secreted signals in distinct genetic settings. Because parabiotic animals share a common circulation, any blood or blood-borne factor from one animal will be exchanged with its partner and vice versa. Thus, cells and molecular factors derived from one genetic background can be studied in the context of a second genetic background. Parabiosis of adult mice has been  used extensively to research aging, cancer, diabetes, obesity, and brain development. More recently, parabiosis of zebrafish embryos has been used to study the developmental biology of hematopoiesis. In contrast to mice, the transparent nature of zebrafish embryos permits the direct visualization of cells in the parabiotic context, making it a uniquely powerful method for investigating fundamental cellular and molecular mechanisms. The utility of this technique, however, is limited by a steep learning curve for generating the parabiotic zebrafish embryos. This protocol provides a step-by-step method on how to surgically fuse the blastulae of two zebrafish embryos of different genetic backgrounds to investigate the role of candidate genes of interest. In addition, the parabiotic zebrafish embryos are tolerant to heat shock, making temporal control of gene expression possible. This method does not require a sophisticated set-up and has broad applications for studying cell migration, fate specification, and differentiation in vivo during embryonic development.

PMID:
27341538
PMCID:
PMC4927786
DOI:
10.3791/54168
[Indexed for MEDLINE]
Free PMC Article

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