In Situ Hybridization: Fruit Fly Embryos and Tissues

Wilk, Ronit; Murthy, Sreenivasa; Yan, Haixu; Krause, Henry M.

doi:10.1002/9780470089941.et0903s04

Cited by 27 publications

(27 citation statements)

References 16 publications

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“…Oregon-R (wild type) embryos were collected, fixed, and hybridized as described in (Wilk et al 2010). All the probes used in this study were produced as described in the same paper.…”

Section: Methodsmentioning

confidence: 99%

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Spatial Profiling of Nuclear Receptor Transcription Patterns over the Course ofDrosophilaDevelopment

Wilk

Krause

2013

G3 Genes|Genomes|Genetics

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Previous work has shown that many of the 18 family members of Drosophila nuclear receptor transcription factors function in a temporal hierarchy to coordinate developmental progression and growth with the rate limiting process of metabolism. To gain further insight into these interactions and processes, we have undertaken a whole-family analysis of nuclear receptor mRNA spatial expression patterns over the entire process of embryogenesis, as well as the 3rd instar wandering larva stage, by using high-resolution fluorescence in situ hybridization. Overall, the patterns of expression are remarkably consistent with previously mapped spatial activity profiles documented during the same time points, with similar hot spots and temporal profiles in endocrine and metabolically important tissues. Among the more remarkable of the findings is that the majority of mRNA expression patterns observed show striking subcellular distributions, indicating potentially critical roles in the control of protein synthesis and subsequent subcellular distributions. These patterns will serve as a useful reference for future studies on the tissue-specific roles and interactions of nuclear receptor proteins, partners, cofactors and ligands.

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“…Oregon-R (wild type) embryos were collected, fixed, and hybridized as described in (Wilk et al 2010). All the probes used in this study were produced as described in the same paper.…”

Section: Methodsmentioning

confidence: 99%

“…Preparation, fixation, and in situ hybridization were performed as described (Wilk et al 2010) with one modification: quenching of endogenous horseradish peroxidase was performed for 30 min in 0.3% H 2 O 2 in PBS. Fresh solution was substituted at 15-min mark.…”

Section: Methodsmentioning

confidence: 99%

Spatial Profiling of Nuclear Receptor Transcription Patterns over the Course ofDrosophilaDevelopment

Wilk

Krause

2013

G3 Genes|Genomes|Genetics

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“…To detect labial mRNA in the eye disc, in situ hybridization was performed using antidigoxigenin-AP (Roche, 11093274910) and NBT-BCIP solution (Sigma, 72091) [94].…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Developmentally regulated autophagy is required for eye formation in Drosophila

et al. 2018

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“…Probe production was described in Lecuyer et al (2008) and Wilk et al (2010). Embryos were imaged as described above.…”

Section: Embryo Staining and Imagingmentioning

confidence: 99%

“…Fluorescent in situ hybridization y w embryos were fixed, and hybridized as described as in Wilk et al (2010) with the probe RE34385 for step (FlyFISH database, http://fly-fish.ccbr.utoronto.ca). Probe production was described in Lecuyer et al (2008) and Wilk et al (2010).…”

Section: Embryo Staining and Imagingmentioning

confidence: 99%

Germ Cell Segregation from the Drosophila Soma Is Controlled by an Inhibitory Threshold Set by the Arf-GEF Steppke

Lee

Wilk

et al. 2015

Genetics

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Germline cells segregate from the soma to maintain their totipotency, but the cellular mechanisms of this segregation are unclear. The Drosophila melanogaster embryo forms a posterior group of primordial germline cells (PGCs) by their division from the syncytial soma. Extended plasma membrane furrows enclose the PGCs in response to the germ plasm protein Germ cell-less (Gcl) and Rho1-actomyosin activity. Recently, we found that loss of the Arf-GEF Steppke (Step) leads to similar Rho1-dependent plasma membrane extensions but from pseudocleavage furrows of the soma. Here, we report that the loss of step also leads to premature formation of a large cell group at the anterior pole of the embryo . These anterior cells lacked germ plasm, but budded and formed at the same time as posterior PGCs, and then divided asynchronously as PGCs also do. With genetic analyses we found that Step normally activates Arf small G proteins and antagonizes Rho1-actomyosin pathways to inhibit anterior cell formation. A uniform distribution of step mRNA around the one-cell embryo cortex suggested that Step restricts cell formation through a global control mechanism. Thus, we examined the effect of Step on PGC formation at the posterior pole. Reducing Gcl or Rho1 levels decreased PGC numbers, but additional step RNAi restored their numbers. Reciprocally, GFP-Step overexpression induced dosage-and Arf-GEF-dependent loss of PGCs, an effect worsened by reducing Gcl or actomyosin pathway activity. We propose that a global distribution of Step normally sets an inhibitory threshold for Rho1 activity to restrict early cell formation to the posterior.

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In Situ Hybridization: Fruit Fly Embryos and Tissues

Cited by 27 publications

References 16 publications

Spatial Profiling of Nuclear Receptor Transcription Patterns over the Course ofDrosophilaDevelopment

Spatial Profiling of Nuclear Receptor Transcription Patterns over the Course ofDrosophilaDevelopment

Developmentally regulated autophagy is required for eye formation in Drosophila

Germ Cell Segregation from the Drosophila Soma Is Controlled by an Inhibitory Threshold Set by the Arf-GEF Steppke

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