Long-term in vivo imaging of Drosophila larvae

Kakanj, Parisa; Eming, Sabine A.; Partridge, Linda; Leptin, Maria

doi:10.1038/s41596-019-0282-z

Cited by 31 publications

(31 citation statements)

References 127 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Imaging of live larva was done on a Nikon Eclipse Ti wide-field microscope using 10x or 20x air objectives. Larva were chosen immediately after shedding their first instar cuticle and were anesthetized using diethyl ether (Kakanj et al, 2020) before being mounted in Vectashield (Vector Labs, Burlingame, CA) with a 0.17mm spacer. The red and green channels in Fig.…”

Section: Softwarementioning

confidence: 99%

Non-canonical Eclosion Hormone-Expressing Cells Regulate Drosophila Ecdysis

et al. 2020

View full text Add to dashboard Cite

Eclosion hormone (EH) was originally identified as a brain-derived hormone capable of inducing the behavioral sequences required for molting across insect species. However, its role in this process (called ecdysis) has since been confounded by discrepancies in the effects of genetic and cellular manipulations of EH function in Drosophila. Although knock-out of the Eh gene results in severe ecdysis-associated deficits accompanied by nearly complete larval lethality, ablation of the only neurons known to express EH (i.e. V m neurons) is only partially lethal and surviving adults emerge, albeit abnormally. Using new tools for sensitively detecting Eh gene expression, we show that EH is more widely expressed than previously thought, both within the nervous system and in somatic tissues, including trachea. Ablating all Eh-expressing cells has effects that closely match those of Eh gene knock-out; developmentally suppressing them severely disrupts eclosion. Our results thus clarify and extend the scope of EH action.

show abstract

Section: Softwarementioning

confidence: 99%

Non-canonical Eclosion Hormone-Expressing Cells Regulate Drosophila Ecdysis

et al. 2020

View full text Add to dashboard Cite

show abstract

“…This reporter is recruited to the plasma membrane by PIP3. As a result, GFP fluorescence is observed on the membrane if PI3K is active in cells expressing this reporter (Britton et al., 2002; Colombani et al., 2003; Kakanj et al., 2020). As an example, we selected cells with Hsc70‐4 silencing based on the simpler method of construction of the fly stocks used in this experiment.…”

Section: Resultsmentioning

confidence: 99%

Heat shock cognate 70 genes contribute to Drosophila spermatocyte growth progression possibly through the insulin signaling pathway

et al. 2021

View full text Add to dashboard Cite

Drosophila spermatocytes grow up to 25 times their original volume before the onset of male meiosis. Several insulin‐like peptides and their cognate receptors (InR) are essential for the cell growth process in Drosophila. Here, we aimed to identify additional signaling pathways and other regulatory factors required for germline cell growth in Drosophila males. Spermatocyte‐specific expression of the dominant‐negative form of InR inhibits cell growth. Conversely, constitutively active forms of signaling factors downstream of InR suppress growth inhibition. Furthermore, hypomorphic mutations in the target of rapamycin (Tor) inhibit spermatocyte growth. These data indicate that the insulin/TOR pathway is essential for the growth of premeiotic spermatocytes. RNA interference (RNAi) screening for the identification of other novel genes associated with cell growth showed that the silencing of each of the five members of heat shock cognate 70 (Hsc70) genes significantly inhibited the process. Hsc70‐silenced spermatocytes showed Akt inhibition downstream of the insulin signaling pathway. Our pleckstrin homology domain‐green fluorescent protein (PH‐GFP) reporter studies indicated that PI3K remained activated in Hsc70‐4‐silenced cells, suggesting that the Hsc70‐4 protein possibly targets Akt or Pdk1 acting downstream of PI3K. Moreover, each of the Hsc70 proteins showed different subcellular localizations. Hsc70‐2 exhibited cytoplasmic colocalization with Akt in spermatocytes before nuclear entry of the kinase during the growth phase. These results indicated the involvement of Hsc70 proteins in the activation of various steps in the insulin signaling pathway, which is essential for spermatocyte growth. Our findings provide insights into the mechanism(s) that enhance signal transduction to stimulate the growth of Drosophila spermatocytes.

show abstract

“…Each larva was placed into an enclosed petri dish filled with evaporated diethyl ether. The larvae remain immobile for several hours after being exposed to diethyl ether for around four minutes (Kakanj et al, 2020). When the larvae were immobile, insect pins were inserted into the head and tail.…”

Section: Methodsmentioning

confidence: 99%

A neuromechanical model for Drosophila larval crawling based on physical measurements

Sun

Liu

et al. 2020

Preprint

View full text Add to dashboard Cite

Animal locomotion requires dynamic interactions between neural circuits, muscles, and surrounding environments. In contrast to intensive studies on neural circuits, the neuromechanical basis for animal behaviour remains unclear due to the lack of information on the physical properties of animals. Here, taking Drosophila larvae as a model system, we proposed an integrated neuromechanical model based on physical measurements. The physical parameters were obtained by a stress-relaxation assay, and the neural circuit motif was extracted from a chain of excitatory and inhibitory interneurons, which was identified previously by connectomics. Based on the model, we systematically performed perturbation analyses on the parameters in the model to study their kinematic effects on locomotion performance. We found that modification of most of the parameters in the simulation could increase the speed of locomotion. Our physical measurement and modelling would provide a new framework for neural circuit studies and soft robot engineering.

show abstract

Long-term in vivo imaging of Drosophila larvae

Cited by 31 publications

References 127 publications

Non-canonical Eclosion Hormone-Expressing Cells Regulate Drosophila Ecdysis

Non-canonical Eclosion Hormone-Expressing Cells Regulate Drosophila Ecdysis

Heat shock cognate 70 genes contribute to Drosophila spermatocyte growth progression possibly through the insulin signaling pathway

A neuromechanical model for Drosophila larval crawling based on physical measurements

Contact Info

Product

Resources

About