Progeny from crosses using the Hand -GAL4 driver appeared in roughly equal ratios Figure 2a , indicating there is no obvious effect of manipulating dTORC1 in cardiac cells. To distinguish between these hypotheses, the level of tTA expression mediated by DJ, DJ or both transgenes was measured across the life span with the tetO-lacZ reporter gene Fig. In this case half the progeny should be knockdown, so the expected ratio is 0. MyoD and the transcriptional control of myogenesis. The loss of repression can be addressed by testing different promoters to drive tTA expression and the timing of the induction may be different for a different GAL4 driver.

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At this age, uninduced expression is additionally seen in the male abdominal muscles. Furthermore we show evidence that the fly Pkd2 mediates muscle contractility via calcium homoeostasis through a functional coupling with the calcium-releasing ryanodine receptor channel on the sarcoplasmic reticulum. While the expression in the digestive system is similar for both transgenes in the pupal and adult stages, expression in the central nervous remains undetectable with DJ At this age, uninduced expression is additionally seen in the male abdominal muscles.

These data suggest that activation of mTORC1 signaling occurs during differentiation, consistent with previous reports You are now also subscribed to the subject areas of this publication and will receive updates in the daily or weekly email digests if turned on. Competing Interests The authors declare there are no competing interests.

The role of TORC1 in muscle development in Drosophila

The x -axis shows the type of Tet-off GAL80 transgene and copy number. Expressed in all embryonic and larval somatic muscles. The UAS-grim lethality test facilitates the determination of the developmental period during which GAL4 activity is not inhibited. PBttk p FLAG rescues Df 1 w67k Pkd2 is located on the second chromosome.


HA rescues Pink1 B9. All raw data, analyzed data and code used to analyze the data and generate figures is available at http: However this driver did not reduce the hatching rate in the UAS-grim assay, which indicates that the driver has little or no expression in the embryo.

To determine the order 24v which myogenic markers are induced during myogenesis, we performed a time course experiment in C2C12 cells.

FlyBase Allele Report: Scer\GAL4[howB]

Assays gal Muscle Functions —All larval assays were repeated in triplicate to obtain standard error bars. The rate of waste excretion depends on food passage along the entire digestive tract and thus is the best measurement for overall contractility of the visceral smooth muscle cells. Although the absolute number of dead pupae was variable among the shRNA- Raptor lines using c -GAL4, in all cases the percentage of dead flies in pupal cases was significantly greater than controls Figure 3g.

This study constructed and characterized Tet-off GAL80 transgenes designed to allow temporal control of GAL4 activity in aging adult muscles.

FlyBase Insertion Report: Dmel\P{GawB}how[24B]

Therefore, DJ was selected as the driverused to test the Tet-off GAL80 transgenes since it has the dual advantage of undetectable expression in muscles during the pre-adult stages 24g high, gwl4 expression in the adult muscles. First, we tested whether inhibition of the dTORC1 pathway affected the development of these flies. Block gao4, promoters; Pentagon, endogenous enhancers; Lozenges, P-element inverted repeats; Triangles, insulator sequences; Ovals, proteins; Rectangles, coding sequences or exons; Rounded rectangles, transcription termination sequences; Striped rectangle, pUC8 plasmid sequence.


Rapamycin blocks C2C12 differentiation. In this case half the progeny should be knockdown, so the expected ratio is 0. Morphologically, there was a complete lack of fused myotubes in the rapamycin treated cells at all time points observed Figure 1e. ADPKD is also a disease of the blood vessels 5.

To measure the rates of waste excretion, newly hatched larvae were fed with bromphenol blue-dyed yeast paste for 4 h, and those that contained the blue food in the hindgut ga,4 then transferred to dye-free food and scored for the complete loss of the blue food in the gut in 1 h. Effects of Pkd2 on skeletal muscle functions. This leaves mouth hook marks on the agar surface.

Knockdown flies are shown in red throughout. To test the genetic interaction between Pkd2 and Ryrwe used the weak Ryr allele. Statistical analyses were performed using the R statistical package version 3. UAS is a suppressor of indirect flight muscle phenotype of park 1.

Essential role of Duox in stabilization of Drosophila wing. In c -GAL4 driven Raptor knockdown flies we observed a critical period of about 20 days after eclosure during which the Raptor knockdown flies are still prone to early death.