A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

Nagarkar-Jaiswal, Sonal; Lee, Pei Tseng; Campbell, Megan E.; Chen, Kuchuan; Anguiano-Zarate, Stephanie S.; Gutierrez, Manuel Cantu; Busby, Theodore; Lin, Wen; He, Yuchun; Schulze, Karen L.; Booth, Benjamin W.; Evans-Holm, Martha; Venken, Koen J. T.; Levis, Robert; Spradling, Allan C.; Hoskins, Roger A.; Bellen, Hugo J.

doi:10.7554/elife.05338

Cited by 363 publications

(398 citation statements)

References 65 publications

Supporting

Mentioning

371

Contrasting

Order By: Relevance

“…S3A). It is interesting to note that deGradFP expression produces a stronger defect than GFP TRiP; a similar result has been observed by Nagarkar-Jaiswal et al (2015).…”

Section: Rab8 Exhibits Dynamic Behaviors In the Early Drosophila Embryosupporting

confidence: 64%

Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster

Mavor

Miao

Zuo

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

One of the most fundamental changes in cell morphology is the ingression of a plasma membrane furrow. The Drosophila embryo undergoes several cycles of rapid furrow ingression during early development that culminate in the formation of an epithelial sheet. Previous studies have demonstrated the requirement for intracellular trafficking pathways in furrow ingression; however, the pathways that link compartmental behaviors with cortical furrow ingression events are unclear. Here, we show that Rab8 has striking dynamic behaviors in vivo. As furrows ingress, cytoplasmic Rab8 puncta are depleted and Rab8 accumulates at the plasma membrane in a location that coincides with known regions of directed membrane addition. We additionally use CRISPR/Cas9 technology to N-terminally tag Rab8, which is then used to address endogenous localization and function. Endogenous Rab8 displays partial coincidence with Rab11 and the Golgi, and this colocalization is enriched during the fast phase of cellularization. When Rab8 function is disrupted, furrow formation in the early embryo is completely abolished. We also demonstrate that Rab8 behaviors require the function of the exocyst complex subunit Sec5 as well as the recycling endosome protein Rab11. Active, GTPlocked Rab8 is primarily associated with dynamic membrane compartments and the plasma membrane, whereas GDP-locked Rab8 forms large cytoplasmic aggregates. These studies suggest a model in which active Rab8 populations direct furrow ingression by guiding the targeted delivery of cytoplasmic membrane stores to the cell surface through interactions with the exocyst tethering complex.

show abstract

“…S3A). It is interesting to note that deGradFP expression produces a stronger defect than GFP TRiP; a similar result has been observed by Nagarkar-Jaiswal et al (2015).…”

Section: Rab8 Exhibits Dynamic Behaviors In the Early Drosophila Embryosupporting

confidence: 64%

Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster

Mavor

Miao

Zuo

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…We used the following fly stocks: Oregon R; Tor A ; Tj-Gal4; UAS-Akt1 RNAi (TRiP HMS00007); UAS-Tor RNAi (TRiP GL00156); UAS-Tsc1 RNAi (TRiP HMJ21477); UAS-Raptor RNAi (TRiP HMS00124); UAS-Pten RNAi (TRiP JF01859); UAS-Dp110; UAS-P35; UAS-ImpL2 (gift of E. Hafen); UAS-ImpL2-RNAi (VDRC #30931); UAS-Dilp6 RNAi (TRiP JF01348); Dilp6-Gal4 (NP1079-GAL4, DGRC Stock Center, Kyoto, Japan); ImpL2-GFP (ImpL2 MI01638-GFSTF.0 ) (Nagarkar-Jaiswal et al, 2015); and tGPH (PH-GFP) (Britton et al, 2002). Tj-Gal4 is expressed in CySCs and in early differentiating cyst cells.…”

Section: Fly Stocks and Husbandrymentioning

confidence: 99%

Somatic stem cell differentiation is regulated by PI3K/Tor signaling in response to local cues

et al. 2016

View full text Add to dashboard Cite

Stem cells reside in niches that provide signals to maintain selfrenewal, and differentiation is viewed as a passive process that depends on loss of access to these signals. Here, we demonstrate that the differentiation of somatic cyst stem cells (CySCs) in the Drosophila testis is actively promoted by PI3K/Tor signaling, as CySCs lacking PI3K/Tor activity cannot differentiate properly. We find that an insulin peptide produced by somatic cells immediately outside of the stem cell niche acts locally to promote somatic differentiation through Insulin-like receptor (InR) activation. These results indicate that there is a local 'differentiation' niche that upregulates PI3K/Tor signaling in the early daughters of CySCs. Finally, we demonstrate that CySCs secrete the Dilp-binding protein ImpL2, the Drosophila homolog of IGFBP7, into the stem cell niche, which blocks InR activation in CySCs. Thus, we show that somatic cell differentiation is controlled by PI3K/Tor signaling downstream of InR and that the local production of positive and negative InR signals regulates the differentiation niche. These results support a model in which leaving the stem cell niche and initiating differentiation are actively induced by signaling.

show abstract

“…w;CCAP-Gal80;+ was described in Luan et al (2006a); w;CCAP-Gal4DBD;+ was described in Luan et al (2006b), and the Trojan exon lines ChaT MI04508 -Gal4DBD, VGlut MI04979 -Gal4DBD, amon MI00899 -p65AD, and Gad1 MI09277 -p65AD were described in Diao et al (2015) All other lines were obtained from the Bloomington Stock Center at Indiana University, including those containing the MI00949 and MI06847 MiMIC insertions in the ETHR gene (Bloomington Stocks 32718 and 41110, respectively), which were generated by the Drosophila Gene Disruption Project, http://flypush.imgen.bcm.tmc.edu/pscreen/mimic.html (Nagarkar-Jaiswal et al 2015). Lk-Gal4 was Bloomington Stock 51993.…”

Section: Fly Linesmentioning

confidence: 99%

The Splice Isoforms of the Drosophila Ecdysis Triggering Hormone Receptor Have Developmentally Distinct Roles

et al. 2015

View full text Add to dashboard Cite

To grow, insects must periodically shed their exoskeletons. This process, called ecdysis, is initiated by the endocrine release of Ecdysis Trigger Hormone (ETH) and has been extensively studied as a model for understanding the hormonal control of behavior. Understanding how ETH regulates ecdysis behavior, however, has been impeded by limited knowledge of the hormone's neuronal targets. An alternatively spliced gene encoding a G-protein-coupled receptor (ETHR) that is activated by ETH has been identified, and several lines of evidence support a role in ecdysis for its A-isoform. The function of a second ETHR isoform (ETHRB) remains unknown. Here we use the recently introduced "Trojan exon" technique to simultaneously mutate the ETHR gene and gain genetic access to the neurons that express its two isoforms. We show that ETHRA and ETHRB are expressed in largely distinct subsets of neurons and that ETHRA-but not ETHRB-expressing neurons are required for ecdysis at all developmental stages. However, both genetic and neuronal manipulations indicate an essential role for ETHRB at pupal and adult, but not larval, ecdysis. We also identify several functionally important subsets of ETHR-expressing neurons including one that coexpresses the peptide Leucokinin and regulates fluid balance to facilitate ecdysis at the pupal stage. The general strategy presented here of using a receptor gene as an entry point for genetic and neuronal manipulations should be useful in establishing patterns of functional connectivity in other hormonally regulated networks.

show abstract

A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

Cited by 363 publications

References 65 publications

Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster

Rab8 directs furrow ingression and membrane addition during epithelial formation in Drosophila melanogaster

Somatic stem cell differentiation is regulated by PI3K/Tor signaling in response to local cues

The Splice Isoforms of the Drosophila Ecdysis Triggering Hormone Receptor Have Developmentally Distinct Roles

Contact Info

Product

Resources

About