This suggests that some early expression of the Gal4-VP16 is due to non-specific ubiquitous activation of the heat-shock promoter during embryogenesis. Optimized Gal4 genetics for permanent gene expression mapping in zebrafish. At 48 hpf, the Gal80 expressing animals did not show any fluorescence, whereas control fish expressed Kaede in scattered neurons Fig. GAL4 is then only expressed in cells where the driver gene is usually active. However, we have found Draper to be dispensable for TotM induction in response to actin injection.

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Animals carrying the hsp: The authors could back off their argument that this study provides evidence for a conserved need for actin detection, especially in light of their lack of evidence for the importance of actin-dependent responses.

Delaying Gal4-VP16 activity with morpholinos There are currently hundreds of stable transgenic lines expressing Gal4-VP16 in various cellular populations in nearly every organ.

We then assessed the resulting fish for green fluorescence and compared them with those injected with the HuC: Temperature-sensitive control of protein activity by conditionally splicing inteins. The following day, the resulting embryos were analyzed for gross anatomical defects and for the expression of green-fluorescent Kaede Fig. Relative expression levels of TotM 24 hr after injection are depicted. Wolbachia- free flies were generated as previously described Chrostek et al.

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The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The improvement of the efficiency of transgenesis in the zebrafish has created the opportunity to generate a variety of transgenic lines bearing Gal4 activators or UAS-driven effectors [11] gwl4, [12][13].


Low levels of Kaede green may be due to incomplete photoconversion, or to Kaede protein translated from mRNA present at the time of photoconversion. Sequenced reads were mapped to Flybase gene set version 6. Detection of F- and total actin by dot blot was carried out as described Ahrens et al.

A tissue-scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. The reviewers have discussed the reviews with one another and the Reviewing Editor has drafted this decision to help you prepare a revised submission.

Red indicates higher expression and blue indicates lower expression relative to the mean expression of probes across all samples.

They also suggest that the gradual release of Gal4 inhibition likely due to the degradation axtin the Gal80 allows the expression of the UAS-driven genes at later stages of development with an average onset at 4 dpf.

Ideally, the authors should provide evidence for the importance of actin signaling in a physiological or pathological setting. Therefore, the simplest interpretation of our data is that Upd3 is synthesised by fat body cells that detect extracellular actin via a sensor s that couple s to a Nox-Src42A-Shark cascade Figure 8—figure supplement 1.

FlyBase Recombinant Construct Report: P{Act5C-GAL4}

To address this question, Srinivasan, Gordon et al. GalMyc] transgenic line is able to inhibit Gal4 activity upon heat shock. In order to quantify the repressive activity of Gal80, we looked at the level of Td-Tomato expression at different time points in cells from fish injected with increasing concentrations of Gal80 RNA Fig. We also validated a strategy to delay Gal4-driven gene expression using a morpholino targeted to Gal4.


The Gal4 protein from yeast is a transcription factor that activates the expression of genes required for growth on galactose. In contrast to Upd3, knockdown of Upd1 and Upd2 in fat body cells had no effect on the response to actin Figure 5econfirming Upd3 as the key cytokine.

Thus, we wanted to devise an alternative strategy to control the timing of Gal4-VP16 activity. The identity of the putative receptor that recognises extracellular actin in Drosophila remains unknown.

Figure 8—figure supplement 1. In conclusion, we have validated two strategies to delay Gal4 activity in the zebrafish.

Decision letter

Scott EK, Baier H. Cold Spring Harbor Perspectives in Biology. Data are representative of at least two independent experiments with duplicate samples. Biochemical and Biophysical Research Communications. Small-molecule regulation of zebrafish gene expression. B Red fluorescence quantification of tdTomato expressing cells.

B Anti-myc western blotting of 48 hpf fish resulting from a cross of Tg[hsp The pEntry vectors containing the TdTomato, the membrane targeting sequence and the HuC promoter have been described previously [32].