Regulatory cocktail for dopaminergic neurons in a protovertebrate identified by whole-embryo single-cell transcriptomics
- Takeo Horie1,2,3,9,
- Ryoko Horie1,2,9,
- Kai Chen2,9,
- Chen Cao2,9,
- Masashi Nakagawa4,
- Takehiro G. Kusakabe5,6,
- Noriyuki Satoh7,
- Yasunori Sasakura1 and
- Michael Levine2,8
- 1Shimoda Marine Research Center, University of Tsukuba, Shimoda, Shizuoka 415-0025, Japan;
- 2Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey 08544, USA;
- 3Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology (PREST), Kawaguchi, Saitama 332-0012, Japan;
- 4Department of Life Science, Graduate School of Life Science, University of Hyogo, Kamigori, Ako-gun, Hyogo 678-1297, Japan;
- 5Department of Biology, Faculty of Science and Engineering, Konan University, Kobe, Hyogo 658-8501, Japan;
- 6Institute for Integrative Neurobiology, Konan University, Kobe, Hyogo 658-8501, Japan;
- 7Marine Genomics Unit, Okinawa Institute of Science and Technology Graduate University, Onna, Okinawa 904-0495, Japan;
- 8Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA
- Corresponding authors: msl2{at}princeton.edu, sasakura{at}shimoda.tsukuba.ac.jp, horie{at}shimoda.tsukuba.ac.jp
-
↵9 These author contributed equally to this work.
Abstract
The CNS of the protovertebrate Ciona intestinalis contains a single cluster of dopaminergic (DA) neurons, the coronet cells, which have been likened to the hypothalamus of vertebrates. Whole-embryo single-cell RNA sequencing (RNA-seq) assays identified Ptf1a as the most strongly expressed cell-specific transcription factor (TF) in DA/coronet cells. Knockdown of Ptf1a activity results in their loss, while misexpression results in the appearance of supernumerary DA/coronet cells. Photoreceptor cells and ependymal cells are the most susceptible to transformation, and both cell types express high levels of Meis. Coexpression of both Ptf1a and Meis caused the wholesale transformation of the entire CNS into DA/coronet cells. We therefore suggest that the reiterative use of functional manipulations and single-cell RNA-seq assays is an effective means for the identification of regulatory cocktails underlying the specification of specific cell identities.
Keywords
Footnotes
-
Supplemental material is available for this article.
-
Article published online ahead of print. Article and publication date are online at http://www.genesdev.org/cgi/doi/10.1101/gad.317669.118.
- Received June 9, 2018.
- Accepted August 2, 2018.
This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
