A Gal4/UAS system for conditional transgene expression in rhombomere 4 of the zebrafish hindbrain
- Authors
- Choe, S.K., Nakamura, M., Ladam, F., Etheridge, L., and Sagerström, C.G.
- ID
- ZDB-PUB-120416-10
- Date
- 2012
- Source
- Developmental Dynamics : an official publication of the American Association of Anatomists 241(6): 1125-1132 (Journal)
- Registered Authors
- Choe, Seong-Kyu, Etheridge, Letitiah, Nakamura, Mako, Sagerström, Charles
- Keywords
- none
- MeSH Terms
-
- Microinjections
- Animals, Genetically Modified/embryology*
- Animals, Genetically Modified/metabolism
- Actins/genetics
- Actins/metabolism*
- Animals
- Trans-Activators/genetics
- Trans-Activators/metabolism*
- DNA Primers/genetics
- Rhombencephalon/embryology*
- Rhombencephalon/metabolism
- Gene Expression Regulation, Developmental/genetics*
- Zebrafish
- Transgenes/genetics*
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism*
- Immunohistochemistry
- Crosses, Genetic
- In Situ Hybridization
- Oligonucleotides/genetics
- PubMed
- 22499412 Full text @ Dev. Dyn.
We first generated the hoxb1a(β-globin):eGFPum8 line that expresses eGFP in hindbrain rhombomere 4 (r4), as well as in facial motor neurons migrating caudally from r4. Second, we generated the hoxb1a(β-globin):Gal4VP16um60 line to express the exogenous Gal4VP16 transcription factor in r4. Lastly, we prepared the UAS(β-actin):hoxa3aum61 line where the hoxa3a gene, which is normally expressed in r5 and r6, is under control of Gal4-regulated UAS elements. Crossing the hoxb1a(β-globin):Gal4VP16um60 line to the UAS(β-actin):hoxa3aum61 line drives robust hoxa3a expression in r4. We find that transgenic expression of hoxa3a in r4 does not affect hoxb1a expression, but has variable effects on migration of facial motorneurons and formation of Mauthner neurons. While cases of somatic transgene silencing have been reported in zebrafish, we have not observed such silencing to date - possibly because of our efforts to minimize repetitive sequences in the transgenic constructs.