Laser-induced gene expression in specific cells of transgenic zebrafish

Mary C. Halloran, Mika Sato-Maeda, James T. Warren, Fengyun Su, Z. Lele, Patrick H. Krone, John Y. Kuwada, Wataru Shoji

Research output: Contribution to journalArticle

345 Citations (Scopus)

Abstract

Over the past few years, a number of studies have described the generation of transgenic lines of zebrafish in which expression of reporters was driven by a variety of promoters. These lines opened up the real possibility that transgenics could be used to complement the genetic analysis of zebrafish development. Transgenic lines in which the expression of genes can be regulated both in space and time would be especially useful. Therefore, we have cloned the zebrafish promoter for the inducible hsp70 gene and made stable transgenic lines of zebrafish that express the reporter green fluorescent protein gene under the control of a hsp70 promoter. At normal temperatures, green fluorescent protein is not detectable in transgenic embryos with the exception of the lens, but is robustly expressed throughout the embryo following an increase in ambient temperature. Furthermore, we have taken advantage of the accessibility and optical clarity of the embryos to express green fluorescent protein in individual cells by focussing a sublethal laser microbeam onto them. The targeted cells appear to develop normally: cells migrate normally, neurons project axons that follow normal pathways, and progenitor cells divide and give rise to normal progeny cells. By generating other transgenic lines in which the hsp70 promoter regulates genes of interest, it should be possible to examine the in vivo activity of the gene products by laser-inducing specific cells to express them in zebrafish embryos. As a first test, we laser-induced single muscle cells to make zebrafish Sema3A1, a semaphorin that is repulsive for specific growth cones, in a hsp70-sema3A1 transgenic line of zebrafish and found that extension by the motor axons was retarded by the induced muscle.

Original languageEnglish
Pages (from-to)1953-1960
Number of pages8
JournalDevelopment
Volume127
Issue number9
Publication statusPublished - May 2000

Fingerprint

Zebrafish
Lasers
Gene Expression
Embryonic Structures
Green Fluorescent Proteins
Genes
Axons
Semaphorins
Growth Cones
Temperature
Muscle Cells
Lenses
Stem Cells
Neurons
Muscles

Keywords

  • Green fluorescent protein
  • Heat-shock gene promoter
  • Semaphorin
  • Transgenic
  • Zebrafish

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Halloran, M. C., Sato-Maeda, M., Warren, J. T., Su, F., Lele, Z., Krone, P. H., ... Shoji, W. (2000). Laser-induced gene expression in specific cells of transgenic zebrafish. Development, 127(9), 1953-1960.

Laser-induced gene expression in specific cells of transgenic zebrafish. / Halloran, Mary C.; Sato-Maeda, Mika; Warren, James T.; Su, Fengyun; Lele, Z.; Krone, Patrick H.; Kuwada, John Y.; Shoji, Wataru.

In: Development, Vol. 127, No. 9, 05.2000, p. 1953-1960.

Research output: Contribution to journalArticle

Halloran, MC, Sato-Maeda, M, Warren, JT, Su, F, Lele, Z, Krone, PH, Kuwada, JY & Shoji, W 2000, 'Laser-induced gene expression in specific cells of transgenic zebrafish', Development, vol. 127, no. 9, pp. 1953-1960.
Halloran MC, Sato-Maeda M, Warren JT, Su F, Lele Z, Krone PH et al. Laser-induced gene expression in specific cells of transgenic zebrafish. Development. 2000 May;127(9):1953-1960.
Halloran, Mary C. ; Sato-Maeda, Mika ; Warren, James T. ; Su, Fengyun ; Lele, Z. ; Krone, Patrick H. ; Kuwada, John Y. ; Shoji, Wataru. / Laser-induced gene expression in specific cells of transgenic zebrafish. In: Development. 2000 ; Vol. 127, No. 9. pp. 1953-1960.
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