ZFIN Image: Burgess et al., 2020, Fig. 4

Image

Figure Caption

Fig. 4 Lineage tracing using ubi:QFGal4-switch. (A) Schematic depicting the organization of a Cre-inducible ubi:QFGal4-switch construct. In the absence of Cre, the ubiquitin B promoter drives expression of mScarletNLS. In the presence of Cre, the loxP-mScarletNLS-loxP expression cassette is excised, resulting in QFGal4 expression. Tg(ubi:QFGal4-switch) transgenic fish can be combined with any existing QUAS reporter and Cre line to create specifically and permanently marked cells for lineage tracing experiments. (B) Fluorescence micrographs showing GFPNLS expression in her4:iCre expressing cells. Epifluorescence micrographs of Tg(ubi:QFGal4-switch)hsc150; Tg(QUAS:GFPNLS)hsc134 double transgenic larvae without Cre (top left panel), or crossed to a Tg(her4.1:iCre)hsc147 transgenic line (bottom left panel). Larvae positive for all three transgenes exhibit robust GFPNLS expression in glial cells of the brain and spinal cord. Right panels: Confocal micrographs showing GFPNLS expression in the brain (coronal section - middle panel) and spinal cord (lateral view - right panel). Note that non-switched cells are marked by mScarletNLS. 4 dpf larvae are shown. (C) Fluorescence micrographs showing GFPNLS expression in foxj1a:iCre expressing cells. Epifluorescence micrographs of Tg(ubi:QFGal4-switch)hsc150; Tg(QUAS:GFPNLS)hsc134 double transgenic larvae crossed to a Tg(foxj1a:iCre) transgenic line and imaged at 5 dpf (left panel). GFPNLS expression is visible along the spinal cord (arrow) and pronephros (arrowhead). Confocal micrographs showing GFPNLS expression in the brain ventricles (lateral view - middle panel) and spinal cord (lateral view - right panel) (Van Gennip et al., 2018).

Figure Data

Construct: Tg(QUASM1-E1B:mScarlet-NLS,myl7:mRFP)

Acknowledgments

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Reprinted from Developmental Biology, 465(2), Burgess, J., Burrows, J.T., Sadhak, R., Chiang, S., Weiss, A., D'Amata, C., Molinaro, A.M., Zhu, S., Long, M., Hu, C., Krause, H.M., Pearson, B.J., An optimized QF-binary expression system for use in zebrafish, 144-156, Copyright (2020) with permission from Elsevier. Full text @ Dev. Biol.