Scanning Electron Microscopy of Drosophila Mutant and Wild Type Eyes

Oster, Irwin I.; Crang, Richard E.

doi:10.2307/3225491

Cited by 4 publications

(3 citation statements)

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“…5D). Their corneal lenses and pseudocones, which are secreted by the cone cells and primary pigment cells, are blurred and irregular, and many of them are fused (Rickenbacher 1954;Oster and Crang 1972;Stumm-Tegethoff and Dicke 1974). In addition, numerous necrotic pits are apparent between an irregular array of ommatidia of variable size.…”

Section: Disrupted Surface Structure and Underlying Cellular Pattern mentioning

confidence: 99%

The Pax2 homolog sparkling is required for development of cone and pigment cells in theDrosophila eye

Fu¹,

Noll²

1997

Genes Dev.

146

142

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A new Drosophila Pax gene, sparkling (spa), implicated in eye development, was isolated and shown to encode the homolog of the vertebrate Pax2, Pax5, and Pax8 proteins. It is expressed in the embryonic nervous system and in cone, primary pigment, and bristle cells of larval and pupal eye discs. In spa pol mutants, a deletion of an enhancer abolishes Spa expression in cone and primary pigment cells and results in a severely disturbed development of non-neuronal ommatidial cells. Spa expression is further required for activation of cut in cone cells and of the Bar locus in primary pigment cells. We suggest close functional analogies between Spa and Pax2 in the development of the insect and vertebrate eye.

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Section: Disrupted Surface Structure and Underlying Cellular Pattern mentioning

confidence: 99%

The Pax2 homolog sparkling is required for development of cone and pigment cells in theDrosophila eye

Fu¹,

Noll²

1997

Genes Dev.

146

142

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“…It is known that the cone cells and primary pigment cells secrete most components of the corneal lens, and secondary pig-ment cells also secrete corneal materials at the boundary between ommatidia (for review, see Charlton-Perkins and Cook, 2010). It has been reported that classical glossy eye fly mutants, such as glass (gl) (Stark and Carlson, 1991), lozenge (lz) (Stark and Carlson, 2000), and sparkling (spa) (Oster and Crang, 1972), possess aberrant "moth-eye" surfaces. Recently, atomic force microscopy (AFM) analysis of glossy eyes overexpressing Wingless showed a dramatic loss of corneal nano-structures (Kryuchkov et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

An RNAi Screen for Genes Involved in Nanoscale Protrusion Formation on Corneal Lens in Drosophila melanogaster

et al. 2016

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The "moth-eye" structure, which is observed on the surface of corneal lens in several insects, supports anti-reflective and self-cleaning functions due to nanoscale protrusions known as corneal nipples. Although the morphology and function of the "moth-eye" structure, are relatively well studied, the mechanism of protrusion formation from cell-secreted substances is unknown. In Drosophila melanogaster, a compound eye consists of approximately 800 facets, the surface of which is formed by the corneal lens with nanoscale protrusions. In the present study, we sought to identify genes involved in "moth-eye" structure, formation in order to elucidate the developmental mechanism of the protrusions in Drosophila. We re-examined the aberrant patterns in classical glossy-eye mutants by scanning electron microscope and classified the aberrant patterns into groups. Next, we screened genes encoding putative structural cuticular proteins and genes involved in cuticular formation using eye specific RNAi silencing methods combined with the Gal4/UAS expression system. We identified 12 of 100 candidate genes, such as cuticular proteins family genes (Cuticular protein 23B and Cuticular protein 49Ah), cuticle secretion-related genes (Syntaxin 1A and Sec61 ββ subunit), ecdysone signaling and biosynthesis-related genes (Ecdysone receptor, Blimp-1, and shroud), and genes involved in cell polarity/cell architecture (Actin 5C, shotgun, armadillo, discs large1, and coracle). Although some of the genes we identified may affect corneal protrusion formation indirectly through general patterning defects in eye formation, these initial findings have encouraged us to more systematically explore the precise mechanisms underlying the formation of nanoscale protrusions in Drosophila.

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“…The structures most frequently examined by SEM include dechorionated embryos and the heads, eyes and thoraces (scutum) of adults (e.g., Harris, 1972;Hodgkin and Bryant, 1978;Oster and Crang 1972;Waterman, 1980;Yun et al, 1994). Processing these stages of Drosophila are relatively straightforward as they have a hard and relatively stable chitinous integument and no significant changes in general SEM protocols are required (Gehring, 2004;Hartman and Hayes, 1971;Jacinto et al, 2000;Stark and Carlson, 2000).…”

Section: Introductionmentioning

confidence: 99%

Processing of soft pupae and uneclosed pharate adults of Drosophila for scanning electron microscopy

Beňo

Liszekova

Farkas

2007

Microscopy Res & Technique

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For over four decades, scanning electron microscopy (SEM) has been used in research involving Drosophila genetics and developmental biology. It allows for observation and documentation of the gross morphology of exoskeletal structures as well as their characterization at very high resolution. In most cases, SEM in Drosophila has been limited to imaging adult heads, thoraces, appendages, and embryos, as these structures are relatively hard and/or easy to process for SEM. In contrast, the structures of the pharate adult stages are difficult to prepare for SEM because their integument is quite soft, they are extremely dirty and they are resistant to the usual processing methods. Here, we present an innovative method to prepare these types of structures. Our protocol efficiently removes extraneous material originating from the exuvial fluid of pharate adults and uses a hydrophobic expansion step to keep the soft exoskeleton of the body inflated. In addition to using immersion fixation, it utilizes fixation within the body that occurs via a reaction between osmium tetroxide and alcohols that are infiltrated into the body during a hydrophobic expansion step. This novel approach results in a properly inflated integument that retains its shape in subsequent procedures. Our method provides a useful, general alternative for processing difficult samples, including soft, biological "whole-mount" specimens and samples that are extremely dirty or resistant to fixative penetration.

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Scanning Electron Microscopy of Drosophila Mutant and Wild Type Eyes

Cited by 4 publications

References 1 publication

The Pax2 homolog sparkling is required for development of cone and pigment cells in theDrosophila eye

The Pax2 homolog sparkling is required for development of cone and pigment cells in theDrosophila eye

An RNAi Screen for Genes Involved in Nanoscale Protrusion Formation on Corneal Lens in Drosophila melanogaster

Processing of soft pupae and uneclosed pharate adults of Drosophila for scanning electron microscopy

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