Abstract
We describe here simple methods for producing transgenic zebrafish reporter lines using BAC clones. The use of BAC clones facilitates creation of useful transgenics as the large amounts of genomic DNA they contain increase the likelihood that reporter gene expression will be properly regulated. Combined with recent advances in live embryo image analysis, this strategy has the potential to greatly advance the investigation of neural cell behavior during development.
Similar content being viewed by others
References
Kimmel CB, Warga RM, Kane DA (1994). Cell cycles and clonal strings during formation of the zebrafish central nervous system. Development 120: 265–276.
Papan C, Campos-Ortega JA (1997). A clonal analysis of spinal cord development in the zebrafish. Dev Genes Evol 207: 71–81.
Papan C, Campos-Ortega JA (1999). Region-specific cell clones in the developing spinal cord of the zebrafish. Dev Genes Evol 209: 135–144.
Eisen JS, Pike SH, Romancier B (1990). An identified motoneuron with variable fates in embryonic zebrafish. J Neurosci 10: 34–43.
Eisen JS (1992). The role of interactions in determining cell fate of two identified motoneurons in the embryonic zebrafish. Neuron 8: 231–240.
Eisen JS (1991). Determination of primary motoneuron identity in developing zebrafish embryos. Science 252: 569–572.
Appel B, Korzh V, Glasgow E, Thor S, Edlund T, Dawid IB, Eisen JS (1995). Motoneuron fate specification revealed by patterned LIM homeobox gene expression in embryonic zebrafish. Development 121: 4117–4125.
Appel B, Givan LA, Eisen JS (2001). Delta-Notch signaling and lateral inhibition in zebrafish spinal cord development. BMC Dev Biol 1: 13.
Bernhardt RR, Patel CK, Wilson SW, Kuwada JY (1992). Axonal trajectories and distribution of GABAergic spinal neurons in wildtype and mutant zebrafish lacking floor plate cells. J Comp Neurol 326: 263–272.
Bernhardt RR, Chitnis AB, Lindamer L, Kuwada JY (1990). Identification of spinal neurons in the embryonic and larval zebrafish. J Comp Neurol 302: 603–616.
Hale ME, Ritter DA, Fetcho JR (2001). A confocal study of spinal interneurons in living larval zebrafish. J Comp Neurol 437: 1–16.
Brand M, Heisenberg CP, Jiang YJ, Beuchle D, Lun K, Furutani-Seiki M, Granato M, Haffter P, Hammerschmidt M, Kane DA, Kelsh RN, Mullins MC, Odenthal J, van Eeden FJ, Nusslein-Volhard C (1996). Mutations in zebrafish genes affecting the formation of the boundary between midbrain and hindbrain. Development 123: 179–190.
Schier AF, Neuhauss SC, Harvey M, Malicki J, Solnica-Krezel L, Stainier DY, Zwartkruis F, Abdelilah S, Stemple DL, Rangini Z, Yang H, Driever W (1996). Mutations affecting the development of the embryonic zebrafish brain. Development 123: 165–178.
Jiang YJ, Brand M, Heisenberg CP, Beuchle D, Furutani-Seiki M, Kelsh RN, Warga RM, Granato M, Haffter P, Hammerschmidt M, Kane DA, Mullins MC, Odenthal J, van Eeden FJ, Nusslein-Volhard C (1996). Mutations affecting neurogenesis and brain morphology in the zebrafish, Danio rerio. Development 123: 205–216.
Udvadia AJ, Linney E (2003). Windows into development: historic, current, and future perspectives on transgenic zebrafish. Dev Biol 256: 1–17.
Koster RW, Fraser SE (2001). Direct imaging of in vivo neuronal migration in the developing cerebellum. Curr Biol 11: 1858–1863.
Gilmour DT, Maischein HM, Nusslein-Volhard C (2002). Migration and function of a glial subtype in the vertebrate peripheral nervous system. Neuron 34: 577–588.
Neumann CJ, Nuesslein-Volhard C (2000). Patterning of the zebrafish retina by a wave of sonic hedgehog activity. Science 289: 2137–2139.
Picker A, Scholpp S, Bohli H, Takeda H, Brand M (2002). A novel positive transcriptional feedback loop in midbrain-hindbrain boundary development is revealed through analysis of the zebrafish pax2.1 promoter in transgenic lines. Development 129: 3227–3239.
Jessen JR, Meng A, McFarlane RJ, Paw BH, Zon LI, Smith GR, Lin S (1998). Modification of bacterial artificial chromosomes through chistimulated homologous recombination and its application in zebrafish transgenesis. Proc Natl Acad Sci USA 95: 5121–5126.
Lee EC, Yu D, Martinez de Velasco J, Tessarollo L, Swing DA, Court DL, Jenkins NA, Copeland NG (2001). A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA. Genomics 73: 56–65.
Westerfield M (2000). The Zebrafish Book. Eugene, Oregon, USA: University of Oregon Press.
Hauptmann G, Gerster T (2000). Multicolor whole-mount in situ hybridization. Methods Mol Biol 137: 139–148.
Yu D, Ellis HM, Lee EC, Jenkins NA, Copeland NG, Court DL (2000). An efficient recombination system for chromosome engineering in Escherichia coli. Proc Natl Acad Sci USA 97: 5978–5983.
Langenberg T, Brand M, Cooper MS (2003). Imaging brain development and organogenesis in zebrafish using immobilized embryonic explants. Developmental Dynamics. In press.
Lu QR, Sun T, Zhu Z, Ma N, Garcia M, Stiles CD, Rowitch DH (2002). Common developmental requirement for Olig function indicates a motor neuron/oligodendrocyte connection. Cell 109: 75–86.
Lu QR, Yuk D, Alberta JA, Zhu Z, Pawlitzky I, Chan J, McMahon AP, Stiles CD, Rowitch DH (2000). Sonic hedgehog - regulated oligodendrocyte lineage genes encoding bHLH proteins in the mammalian central nervous system. Neuron 25: 317–329.
Mizuguchi R, Sugimori M, Takebayashi H, Kosako H, Nagao M, Yoshida S, Nabeshima Y, Shimamura K, Nakafuku M (2001). Combinatorial roles of olig2 and neurogenin2 in the coordinated induction of pan-neuronal and subtype-specific properties of motoneurons. Neuron 31: 757–771.
Novitch BG, Chen AI, Jessell TM (2001). Coordinate regulation of motor neuron subtype identity and pan-neuronal properties by the bHLH repressor Olig2. Neuron 31: 773–789.
Park H, Mehta A, Richardson JS, Appel B (2002). olig2 is required for zebrafish primary motor neuron and oligodendrocyte development. Developmental Biology 248: 356–368.
Sun T, Echelard Y, Lu R, Yuk D, Kaing S, Stiles CD, Rowitch DH (2001). Olig bHLH proteins interact with homeodomain proteins to regulate cell fate acquisition in progenitors of the ventral neural tube. Curr Biol 11: 1413–1420.
Takebayashi H, Nabeshima Y, Yoshida S, Chisaka O, Ikenaka K (2002). The basic helix-loop-helix factor olig2 is essential for the development of motoneuron and oligodendrocyte lineages. Curr Biol 12: 1157–1163.
Takebayashi H, Yoshida S, Sugimori M, Kosako H, Kominami R, Nakafuku M, Nabeshima Y (2000). Dynamic expression of basic helix-loop-helix Olig family members: implication of Olig2 in neuron and oligodendrocyte differentiation and identification of a new member, Olig3. Mech Dev 99: 143–148.
Zhou Q, Wang S, Anderson DJ (2000). Identification of a novel family of oligodendrocyte lineage-specific basic helix-loop-helix transcription factors. Neuron 25: 331–343.
Zhou Q, Choi G, Anderson DJ (2001). The bHLH transcription factor Olig2 promotes oligodendrocyte differentiation in collaboration with Nkx2.2. Neuron 31: 791–807.
Zhou Q, Anderson DJ (2002). The bHLH transcription factors OLIG2 and OLIG1 couple neuronal and glial subtype specification. Cell 109: 61–73.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shin, J., Park, HC., Topczewska, J.M. et al. Neural cell fate analysis in zebrafish using olig2 BAC transgenics. Methods Cell Sci 25, 7–14 (2003). https://doi.org/10.1023/B:MICS.0000006847.09037.3a
Issue Date:
DOI: https://doi.org/10.1023/B:MICS.0000006847.09037.3a