Live imaging of innate immune and preneoplastic cell interactions using an inducible Gal4/UAS expression system in larval zebrafish skin.
Here we describe a method to conditionally induce epithelial cell transformation by the use of the 4-Hydroxytamoxifen (4-OHT) inducible KalTA4-ERT2/UAS expression system(1) in zebrafish larvae, and the subsequent live imaging of innate immune cell interaction with HRASG12V expressing skin cells. The KalTA4-ERT2/UAS system is both inducible and reversible which allows us to induce cell transformation with precise temporal/spatial resolution in vivo. This provides us with a unique opportunity to live image how individual preneoplastic cells interact with host tissues as soon as they emerge, then follow their progression as well as regression. Recent studies in zebrafish larvae have shown a trophic function of innate immunity in the earliest stages of tumorigenesis(2,3). Our inducible system would allow us to live image the onset of cellular transformation and the subsequent host response, which may lead to important insights on the underlying mechanisms for the regulation of oncogenic trophic inflammatory responses. We also discuss how one might adapt our protocol to achieve temporal and spatial control of ectopic gene expression in any tissue of interest.
MRC Centre for Inflammation Research, Queen's Medical Research Institute, The University of Edinburgh.
Thomas
T
Ramezani
MRC Centre for Inflammation Research, Queen's Medical Research Institute, The University of Edinburgh.
Derek W
DW
Laux
MRC Centre for Inflammation Research, Queen's Medical Research Institute, The University of Edinburgh.
Isabel R
IR
Bravo
Department of Cell & Developmental Biology, University College London.
Masazumi
M
Tada
MRC Centre for Inflammation Research, Queen's Medical Research Institute, The University of Edinburgh; yi.feng@ed.ac.uk.
Yi
Y
Feng
D000818:Animals; D030801:Animals, Genetically Modified; D002450:Cell Communication / Q000502:physiology; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000276:immunology* / Q000473:pathology*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D007113:Immunity, Innate; D007814:Larva / Q000502:physiology; D008845:Microinjections; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D011230:Precancerous Conditions / Q000235:genetics / Q000276:immunology* / Q000473:pathology*; D012867:Skin / Q000276:immunology* / Q000473:pathology*; D012879:Skin Physiological Phenomena / Q000235:genetics / Q000276:immunology; D019076:Transgenes; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics
D029961:Zebrafish Proteins; D004247:DNA