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In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences
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Developmental Biology
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JoVE Journal Developmental Biology
In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

DOI:
10.3791/55490-v

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10:36 min

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March 22, 2017

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, , ,
1School of Biosciences,The University of Melbourne, 2Australian Regenerative Medicine Institute (ARMI),Monash University, 3The David J Apple Center for Vision Research, Department of Ophthalmology,Heidelberg University

Chapters

  • 00:05Title
  • 00:52Embryo Microinjection
  • 02:53Preparation for Transplantation
  • 04:59Chimera Generation via Blastula Transplantation
  • 06:37Mounting for Live Imaging
  • 08:27Results: Transgene Expression in Retinal Progenitors from Donor Cells
  • 09:34Conclusion

Summary

Automatic Translation

Automatic Translation

Live tracking of individual WT retinal progenitors in distinct genetic backgrounds allows for the assessment of the contribution of cell non-autonomous signaling during neurogenesis. Here, a combination of gene knockdown, chimera generation via embryo transplantation and in vivo time-lapse confocal imaging was utilized for this purpose.

Tags

In Vivo ImagingTransgenic Gene ExpressionRetinal Progenitor CellsChimeric Zebrafish EmbryosCell Non-autonomous InfluencesFate SpecificationMicroinjectionH2AGFPH2BRFPMRNAFluorescence Microscopy

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