Visualization of Gene Expression Patterns by In Situ Hybridization on Early Stages of Development of Xenopus laevis

2018 ◽  
pp. 325-335
Author(s):  
Heithem M. El-Hodiri ◽  
Lisa E. Kelly
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Stages Of Development ◽  
Early Stages

scholarly journals RNA In Situ Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of Drosophila Species

2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Drosophila Species ◽  
Gene Expression Patterns ◽  
Probe Design ◽  
Tissue Preparation ◽  
Design And Synthesis ◽  
Rna In Situ Hybridization

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

Determination of Gene Expression Patterns by Whole-Mount In Situ Hybridization

2012 ◽  
pp. 15-22 ◽  
Author(s):  
Sergiy Kyryachenko ◽  
Kateryna Kyrylkova ◽  
Mark Leid ◽  
Chrissa Kioussi
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Whole Mount

ZebraFISH: Fluorescent In Situ Hybridization Protocol and Three-Dimensional Imaging of Gene Expression Patterns

Zebrafish ◽  
2006 ◽  
Vol 3 (4) ◽  
pp. 465-476 ◽  
Author(s):  
Monique C.M. Welten ◽  
Simon B. de Haan ◽  
Niels van den Boogert ◽  
Jasprien N. Noordermeer ◽  
Gerda E.M. Lamers ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Expression Patterns ◽  
Three Dimensional ◽  
Gene Expression Patterns ◽  
Three Dimensional Imaging ◽  
Hybridization Protocol

Determination of gene expression patterns using in situ hybridization to Drosophila testes

2009 ◽  
Vol 4 (12) ◽  
pp. 1807-1819 ◽  
Author(s):  
Ceri A Morris ◽  
Elizabeth Benson ◽  
Helen White-Cooper
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Gene Expression Patterns

scholarly journals Extraction and comparison of gene expression patterns from 2D RNA in situ hybridization images

2009 ◽  
Vol 26 (6) ◽  
pp. 761-769 ◽  
Author(s):  
Daniel L. Mace ◽  
Nicole Varnado ◽  
Weiping Zhang ◽  
Erwin Frise ◽  
Uwe Ohler
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Rna In Situ Hybridization

scholarly journals Robustness of the Dorsal morphogen gradient with respect to morphogen dosage

2019 ◽  
Author(s):  
Hadel Al Asafen ◽  
Prasad U. Bandodkar ◽  
Sophia Carrell-Noel ◽  
Gregory T. Reeves
Keyword(s):  
Gene Expression ◽  
Expression Patterns ◽  
Positional Information ◽  
The Body ◽  
Morphogen Gradient ◽  
Gene Expression Patterns ◽  
Facilitated Diffusion ◽  
Uniform Field ◽  
Stages Of Development ◽  
Early Stages

AbstractIn multicellular organisms, the timing and placement of gene expression in a developing tissue assigns the fate of each cell in the embryo in order for a uniform field of cells to differentiate into a reproducible pattern of organs and tissues. This positional information is often achieved through the action of spatial gradients of morphogens. Spatial patterns of gene expression are paradoxically robust to variations in morphogen dosage, given that, by definition, gene expression must be sensitive to morphogen concentration. In this work we investigate the robustness of the Dorsal/NF-κB signaling module with respect to perturbations to the dosage of maternally-expressed dorsal mRNA. The Dorsal morphogen gradient patterns the dorsal-ventral axis of the early Drosophila embryo, and we found that an empirical description of the Dorsal gradient is highly sensitive to maternal dorsal dosage. In contrast, we found experimentally that gene expression patterns are highly robust. Although the components of this signaling module have been characterized in detail, how their function is integrated to produce robust gene expression patterns to variations in the dorsal maternal dosage is still unclear. Therefore, we analyzed a mechanistic model of the Dorsal signaling module and found that Cactus, a cytoplasmic inhibitor for Dorsal, must be present in the nucleus for the system to be robust. Furthermore, active Toll, the receptor that dissociates Cactus from Dorsal, must be saturated. Finally, the vast majority of robust descriptions of the system require facilitated diffusion of Dorsal by Cactus. Each of these three recently-discovered mechanisms of the Dorsal module are critical for robustness. Our work highlights the need for quantitative understanding of biophysical mechanisms of morphogen gradients in order to understand emergent phenotypes, such as robustness.Author SummaryThe early stages of development of an embryo are crucial for laying the foundation of the body plan. The blueprint of this plan is encoded in long-range spatial protein gradients called morphogens. This positional information is then interpreted by nuclei that begin to differentiate by expressing different genes. In fruit fly embryos, the Dorsal morphogen forms a gradient along the dorsal-ventral axis, with a maximum at the ventral midline. This gradient, and the resulting gene expression patterns are extraordinarily robust to variations in developmental conditions, even during early stages of development. Since positional information is interpreted in terms of concentration of the morphogen, one would expect that doubling or halving dosage would result in disastrous consequences for the embryo. However, we observed that development remains robust. We quantified the effect of dosage by experimentally measuring the boundaries of 2 genes, - sna and sog, expressed along the DV axis and found that variation in the boundaries of these genes was minimal, across embryos with different dosages of Dl. We then used a mathematical model to discern components of the Dl system responsible for buffering the effects of dosage and found three specific mechanisms – deconvolution, Toll saturation and shuttling

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