Gustavo A. Cerda and Murray Hargrave contributed equally to this work.
RNA profiling of FAC-sorted neurons from the developing zebrafish spinal cord
Article first published online: 18 DEC 2008
Copyright © 2008 Wiley-Liss, Inc.
Volume 238, Issue 1, pages 150–161, January 2009
How to Cite
Cerda, G. A., Hargrave, M. and Lewis, K. E. (2009), RNA profiling of FAC-sorted neurons from the developing zebrafish spinal cord. Dev. Dyn., 238: 150–161. doi: 10.1002/dvdy.21818
- Issue published online: 18 DEC 2008
- Article first published online: 18 DEC 2008
- Manuscript Accepted: 27 OCT 2008
- Medical Research Council. Grant Number: G0600877
- Royal Society University Research Fellowship
In this report, we describe a successful protocol for isolating and expression-profiling live fluorescent-protein-labelled neurons from zebrafish embryos. As a proof-of-principle for this method, we FAC-sorted and RNA-profiled GFP-labelled spinal CiA interneurons and compared the expression profile of these cells to those of post-mitotic spinal neurons in general and to all trunk cells. We show that RNA of sufficient quality and quantity to uncover both expected and novel transcription profiles via Affymetrix microarray analysis can be extracted from 5,700 to 20,000 FAC-sorted cells. As part of this study, we also further confirm the genetic homology of mammalian and zebrafish V1 interneurons, by demonstrating that zebrafish V1 cells (CiAs) express genes that encode for the transcription factors Lhx1a and Lhx5. This protocol for dissociating, sorting and RNA-profiling neurons from organogenesis-stage zebrafish embryos should also be applicable to other developing organs and tissues and potentially other model organisms. Developmental Dynamics 238:150–161, 2009. © 2008 Wiley-Liss, Inc.