Generating chimeric zebrafish embryos by transplantation.

Kemp, Hilary A and Carmany-Rampey, Amanda and Moens, Cecilia (2009) Generating chimeric zebrafish embryos by transplantation. Journal of visualized experiments : JoVE (29). p. 1394. ISSN 1940-087X

Preview	Text (Complete article) JoVE_Protocol_1394.pdf Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (361kB) | Preview
	Video (Protocol Video) 1394_Moens_F3.mp4 Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (138MB)

Abstract

One of the most powerful tools used to gain insight into complex developmental processes is the analysis of chimeric embryos. A chimera is defined as an organism that contains cells from more than one animal; mosaics are one type of chimera in which cells from more than one genotype are mixed, usually wild-type and mutant. In the zebrafish, chimeras can be readily made by transplantation of cells from a donor embryo into a host embryo at the appropriate embryonic stage. Labeled donor cells are generated by injection of a lineage marker, such as a fluorescent dye, into the one-cell stage embryo. Labeled donor cells are removed from donor embryos and introduced into unlabeled host embryos using an oil-controlled glass pipette mounted on either a compound or dissecting microscope. Donor cells can in some cases be targeted to a specific region or tissue of the developing blastula or gastrula stage host embryo by choosing a transplantation site in the host embryo based on well-established fate maps.

Item Type:	Article or Abstract
DOI:	10.3791/1394
PubMed ID:	19617875
NIHMSID:	NIHMS137341
PMCID:	PMC2770904
Grant Numbers:	R01 HD037909
Keywords or MeSH Headings:	* Animals * Blastula/transplantation * Cell Transplantation/methods* * Embryo Transfer * Embryo, Nonmammalian/cytology * Female * Gastrula/transplantation * Transplantation Chimera/embryology* * Zebrafish/embryology* * Zebrafish/genetics
Subjects:	Therapeutics > Transplantation Organisms > Model organisms Cellular and Organismal Processes > Development
Depositing User:	Library Staff
Date Deposited:	03 Aug 2009 23:20
Last Modified:	14 Feb 2012 14:42
URI:	http://authors.fhcrc.org/id/eprint/322

Repository Administrators Only

View Item