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Imaginal Discs

Q: What are the different types or variations of Imaginal Discs?

Imaginal discs can be categorized into different types based on the adult structures they give rise to. The main types include wing discs, leg discs, eye-antehnna discs, and genital discs. Each type of imaginal disc is responsible for the formation of a specific set of adult features, and they undergo unique developmental processes to achieve this.

Imaginal Discs are specialized groups of cells found in the larvae of holometabolous insects, such as flies, beetles, and moths.
These discs are the precursors to the adult structures, including wings, legs, antennae, and other external features.
During metamorphosis, the imaginal discs undergo rapid growth and differentiation to form the adult form.
Understandinig the development and regulation of imaginal discs is crucial for studyinq insect morphology, physiology, and evolutionary biology.

Most cited protocols related to «Imaginal Discs»

Dissection and Immunofluorescence of Drosophila Tissues

Third instar wandering larvae were collected in ice-cold PBS and the salivary glands, wing imaginal discs, ovaries and fat body dissected. All were fixed in 4% formaldehyde in PBS. Glands, fat body and ovaries were washed in PBS containing 0.3% Triton X-100 (PBST) before blocking in PBST plus 20% FBS followed by primary antibodies in the same buffer. Discs were washed in PBS with 0.05% (v/v) Triton X-100 and blocked in PBX250 [PBS, 0.05% (v/v) Triton X-100, 1 mg/ml BSA, 250 mM NaCl], with antibodies added in PBX buffer [PBS, 0.05% (v/v) Triton X-100, 1 mg/ml BSA]. All were incubated overnight at 4°C with agitation before washing and detection using species-specific Alexa-labeled secondary antibodies (Molecular Probes), and mounting in VECTASHIELD (Vector Laboratories) for confocal microscopy.

Riedel F., Gillingham A.K., Rosa-Ferreira C., Galindo A, & Munro S. (2016). An antibody toolkit for the study of membrane traffic in Drosophila melanogaster. Biology Open, 5(7), 987-992.

Publication 2016

Antibodies Buffers Cloning Vectors Common Cold Fat Body Formaldehyde Imaginal Discs Larva Microscopy, Confocal Molecular Probes Ovary Salivary Glands Sodium Chloride Triton X-100

Genetic Dissection of miR-8 Regulation in Drosophila Neuromuscular Junction

All stocks were maintained and crossed at 25°C according to standard procedures. Stocks were obtained from the Bloomington Stock Center unless otherwise specified. The following loss-of-function alleles and transgenic lines were used: GMR-Yan^Act (ref. 15 (link), 26 (link)) was a gift from R.W. Carthew, UAS-FP₄-mitoEGFP (ref. 23 (link)) was a gift from M. Peifer, ena²¹⁰ and UAS-ena (ref. 27 (link)) were gifts from F. M. Hoffmann, miR-9a^J22 and miR-9a^E39 (ref. 14 (link)) was a gift from F.B. Gao; miR-8-GFP Sensor (ref. 21 (link)) was a gift from H. Ruohola-Baker, EP-atro, and P{EPgy2}GugEY14339. miR-8^Δ2 (ref. 16 (link)) was a gift from S. Cohen; an independent miR-8 null allele ΔmiR-8 was generated as part of a separate study in our laboratory (C.S.L., C.M.L. and D.VV. unpublished observations), tubulinEGFP nerfin-1 3'UTR reporter^{12 (link)} was a gift from J. Brennecke. For the analysis of miR-9a activity in wing imaginal discs, the tubulinEGFP nerfin-1 3'UTR reporter was recombined with ptc-Gal4 on the second autosomal chromosome. The following Gal4 drivers were used to drive ubiquitous, eye, leg, wing disc, pan-neural and mesodermal expression: tubulin-Gal4, GMR-Gal4 and eyeless-Gal4, Dll-Gal4, ptc-Gal4, elav-Gal4 and how^24B-Gal4 respectively. The miR-8-Gal4 line was obtained from the Drosophila Genetic Resource Center (DGRC) and used to drive expression of UAS-CD8GFP.
To generate the allelic combination of ena heterozygous/ΔmiR-8 homozygous genetic background, the ena²¹⁰ allele was recombined with ΔmiR-8 allele on the second autosomal chromosome and the miR-8 NMJ phenotypes were assessed as described above. To test the effect of postsynaptic Ena inhibition on ΔmiR-8 induced NMJ phenotype, UAS-FP₄-mito was expressed using the how^24B-Gal4 driver, in a ΔmiR-8 homozygous mutant background. The specificity of the UAS-FP₄-mito has been previously described^{23 (link)}.

Loya C.M., Lu C.S., Van Vactor D, & Fulga T.A. (2009). Transgenic microRNA inhibition with spatiotemporal specificity in intact organisms. Nature methods, 6(12), 897-903.

Publication 2009

3' Untranslated Regions Alleles Animals, Transgenic Chromosomes Drosophila Genetic Background Gifts Heterozygote Homozygote Imaginal Discs Mesoderm Mitomycin Nervousness Phenotype Psychological Inhibition Tubulin

Genetic Dissection of miR-8 Regulation in Drosophila Neuromuscular Junction

Loya C.M., Lu C.S., Van Vactor D, & Fulga T.A. (2009). Transgenic microRNA inhibition with spatiotemporal specificity in intact organisms. Nature methods, 6(12), 897-903.

Publication 2009

Drosophila Strain Characterization and Lifespan Assay

Drosophila strains.
All the target transgenes
for over-expression (Table 1) were obtained from Bloomington Drosophila Stock Center. The ubiquitous Geneswitch driver lines Act-GS-255B and
Act-GS-255A contain multiple copies of a P element construct in which
expression of the Geneswitch cDNA is under the control of the tissue-general Actin5C promoter [16 (link)]. The UAS-ultraGFP strain contain multiple copies of a UAS-eGFP
construct, and its construction and characterization have been recently
described [18 ]. The Geneswitch system drivers Elav-GS, MHC-GS, S₁-32
and S₁-106 were generously provided by T. Osterwalder and R. Davis
[19 (link),20 (link)].

Drosophila culture.
Drosophila culture and life span assays were performed as described previously [16 (link)].
GeneSwitch virgins were used in the crosses with males of other lines, with the
exception of strains in which the target transgene for over-expression was on
the X chromosome. Life span assays consisted of ~25 flies per vial, and a total
5 vials for each cohort. For survival assays performed at 25^oC,
flies were transferred to new vials ever other day. For survival assays
preformed at 29^oC flies were transferred to new vials every other
day during the first 30-40 days, and then every day for the remainder of the
life span. RU486 (Mifepristone, Sigma) was dissolved in ethanol (100%) to make
a stock solution of 3.2mg/ml. For adult feeding, 50ul RU486 stock solution was
added to the surface of each vial to produce a final concentration of
~160ug/ml; 50ul ethanol was added to the control vials. For larval feeding,
0.5ml of 3.2mg/ml RU486 stock solution (or the indicated diluted concentration)
was added to the surface of each bottle to produce a final concentration of
~160ug/ml (or indicated diluted concentration); 0.5ml ethanol was added to
control bottles.

GeneSwitch driver characterization
. Adult
flies were cultured in vials in the presence and absence of drug for two weeks
prior to dissection. Adult male and female flies, head in half, body in half,
midgut and hindgut, ovary and testes, were photographed. Larvae at 1^st instar, 2^nd instar and 3^rd instar, as well as 3^rd instar dissected tissues (brain, midgut and hindgut, salivary gland, imaginal
discs, and fat body) were also photographed. The Leica MZ FLIII fluorescence
stereomicroscope together with the SPOT software were used for photographs: The
GFP pictures were taken under the fluorescent light with exposure time 4 sec
and a gain of 2.

Statistical analysis.
Mean, standard deviation,
median, percent change in mean, percent change in median, and log rank p value were calculated using R 2.6.2 [58 ]. Analysis of mortality rate was
performed with the WinModest statistical package [59 ]. In the
Gompertz-Makeham model, the increaseof mortality (μ_x)
with age (x) is expressed as: μ_x= ae^bx+c,
where the constant a is the initial mortalityrate, b is the rate of exponential increase in mortality, and c is the
age-independent mortality. The age specific mortality rate (μ_x)
was calculated using WinModest by binning the days over which deaths
were counted (since fly deaths were recorded every other day) such that μ_x = (-ln(N_{x +}_δ_x / N_x )) / δ_x(or P_x = N_{x +}_δ_x / N_x and μ_x = -1/δ_x ln(P_x )), where N_xis the number of flies alive at day x and δ_x is the bin size [2 (link)].
Parameters (a, b, c) were also calculated based on a likelihood ratio
test. The full model (ae^bx+c) was plotted, and the
Gompertz-only component (ae^bx) was used to build the
decomposed survival curves, using μ_x: μ_x= ae^bx, P_x= e^-μx.
For the decomposed survival curves, any value below 0.5% survival was
considered to be the final data point.

Shen J., Curtis C., Tavaré S, & Tower J. (2009). A screen of apoptosis and senescence regulatory genes for life span effects when over-expressed in Drosophila. Aging, 1(2), 191-211.

Publication 2009

Adult Biological Assay Brain Diptera Dissection DNA, Complementary Drosophila Ethanol Fat Body Females Fluorescence Head Human Body Imaginal Discs Larva Light Males Mifepristone Ovary Pharmaceutical Preparations R-38486 Salivary Glands Strains Testis Tissues Transgenes X Chromosome

Visualizing Drosophila Eye Development

Adult lenses were visualized by SEM using anesthetized flies mounted on carbon tabs and directly analyzing them with a Hitachi S-3400N. For thin sections, eyes were fixed for 15 minutes in 4% paraformaldehyde/phosphate-buffered saline, washed twice with 0.1% Triton X/phosphate-buffered saline (PBT), serially dehydrated in ethanol, incubated in 1:1 ethanol/LR-White resin (Electron Microscopy Sciences, Hatfield, PA, USA) for 1 hour and 100% resin for 1 hour, then polymerized with one drop per milliliter of accelerator. Sections (2 μm) were dried for 15 minutes on a slide warmer and stained with 1% toluidine blue for 10 minutes and mounted in Entellan (EMS), or rehydrated in PBT overnight followed by antibody staining and mounting as previously described [74 (link)]. Imaginal discs and retinas dissected from pupa 45 hours after puparium formation (45% pupation) were immunostained as previously described for whole mount adult retinas [74 (link)]. Antibodies were from the Developmental Studies Hybridoma Bank unless indicated otherwise, and diluted as follows: Cut (mouse, 1:100), Pros (rabbit [50 (link)], 1:1,000), Pros (mouse, 1:10), Pros (guinea pig, this paper, 1:1,500), dPax2 (rabbit [28 (link)], 1:50; rabbit, this paper, 1:1500; guinea pig, this paper, 1;1,500), Elav (mouse or rat, 1:200), Eya (mouse, 1:50), BarH1 (rat, this paper, 1:200; rabbit [38 (link)], 1:50), Dlg (mouse, 1:50), E-cadherin (rat, 1:20; rabbit, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA, 1:50), GFP (chicken, Abcam, Cambridge, MA, USA, 1:500), β-gal (chicken, Sigma-Aldrich, St. Louis, MO, USA, 1:1,000), Otd (guinea pig [74 (link)], 1:750), Rh3 (chicken [32 (link)], 1:40), Rh4 (rabbit, gift from C Zuker/N Colley, 1:150), Delta (mouse, 1:50), E(spl) (mouse [44 (link)], 1:1), pERK (rabbit, Sigma-Aldrich, 1:20), m(Espl) (1:2 [44 (link)]) and N-Cad (rat, 1:20). Secondary antibodies were conjugated to Alexa Fluor 488, 555, 647 and 750 nm (goat, Invitrogen) or Cy2, 3 or 5 (donkey, Jackson Immunoresearch, West Grove, PA, USA), and diluted 1:500. Polymerized actin was detected using AlexFluor 488-conjugated phalloidin (Invitrogen), which was added to the secondary antibody dilutions at 1:20 according to the manufacturer's suggestion. Samples were imaged with a Zeiss Apotome, deconvolved with Axiovision 4.6 or Zeiss LSM 700 confocal and processed in Adobe Photoshop 7.0.

Charlton-Perkins M., Whitaker S.L., Fei Y., Xie B., Li-Kroeger D., Gebelein B, & Cook T. (2011). Prospero and Pax2 combinatorially control neural cell fate decisions by modulating Ras- and Notch-dependent signaling. Neural Development, 6, 20.

Publication 2011

Adult alexa fluor 488 Antibodies Carbon Cavia CDH1 protein, human Chickens Diptera Electron Microscopy entellan Equus asinus Ethanol Eye F-Actin Goat Hybridomas Imaginal Discs Immunoglobulins Lens, Crystalline LR white Mice, House Microtomy paraform Phalloidine Phosphates Proline Pupa Rabbits Resins, Plant Retina Saline Solution Technique, Dilution Tolonium Chloride

Most recents protocols related to «Imaginal Discs»

Quantifying Sexual Dimorphism in Larval Drosophila

We used a molecular method. βtubulin85D (FBgn0003889) is reported to be expressed specifically in testes [73 (link)] predicting a dimorphic pattern of βtub85D expression, high in males and low in female larvae. We first verified this by sexing a subset of larvae by eye on the basis of the larger testes versus ovary imaginal discs. We then quantified the level of βtub85D expression in the same larvae, normalized to reference genes elF1A (FBgn0026250) and αTub84B (FBgn0003884) (electronic supplementary material, table S2). As expected, larvae sexed as males had high βtub85D expression and larvae sexed as females had no βtub85D expression above background noise. Thus we used βtub85D expression assays to identify the sex of experimental larvae.

Bennett-Keki S., Fowler E.K., Folkes L., Moxon S, & Chapman T. (2023). Sex-biased gene expression in nutrient-sensing pathways. Proceedings of the Royal Society B: Biological Sciences, 290(1994), 20222086.

Publication 2023

Biological Assay Females Genes Imaginal Discs Larva Males Ovary Testis

Staged Embryo Collection and Imaging

For staged samples, embryos were collected for 2 h intervals as described before^{69 (link)} and allowed to develop at 25 °C until the desired developmental stage (MMP2 knockdown experiments in Fig. 6b–i were performed at 29 °C due to increased efficiency of knockdown). Wing discs were isolated at defined time intervals after egg laying (hAEL). For 72hAEL and older time points, only male larvae were included (positive selection by the transparent genitalia disc well visible in the posterior half of male larvae); 65hAEL data contains male and female larvae since at this time point the genitalia disc is not yet clearly visible.
All larvae of one experiment were dissected, processed and imaged in parallel, using identical solutions in order to reduce experimental variations. Immunostaining of imaginal discs was performed as described previously^{69 (link)}. Discs were mounted in Vectashield Plus (H-1900, Vector Laboratories) using double-sided tape as spacers (TESA 05338) to maintain tissue morphology and avoid squishing of the sample.
All fixed samples were imaged on a Leica SP8 confocal microscope (Leica Application Suite X 3.55.19976) using a ×40 or a ×63/1.4 NA oil-immersion objective. All image stacks of one experiment were acquired in the same session using identical imaging settings. Imaging conditions were chosen to be well within the dynamic range of the fluorescent signal obtained. For optical cross-sections of wing discs stacks with high resolution along the z axis were acquired (typically 0.33 µm of spacing between slices).

Harmansa S., Erlich A., Eloy C., Zurlo G, & Lecuit T. (2023). Growth anisotropy of the extracellular matrix shapes a developing organ. Nature Communications, 14, 1220.

Publication 2023

Cloning Vectors Embryo Epistropheus Females Genitalia Imaginal Discs Larva Males Microscopy, Confocal MMP2 protein, human Submersion Tissues Vision

Quantification of Imaginal Disc Clones

Wing imaginal discs were dissected from third instar wandering stage larvae in PBS. Wing discs were fixed at room temperature for 20 min with 4% formaldehyde in PBS. After washing three times for 5 min with PBS, discs were stained with Hoechst 33,258 (1 µg/ml) for 10 min. After an additional three washes with PBS for 5 min each, discs were mounted in Vectashield Antifade Mounting Medium (Vector Laboratories, #H-1000). An Olympus FluoView 1000 laser-scanning confocal microscope was used for imaging with a 40 × /1.3 oil objective. Ten optical sections with a spacing of 1 µm were acquired with a Kalman filter setting of three.
ImageJ was used for quantification of the clone areas. The z-slice that contained a maximal number of nuclei in the flat wing pouch region was identified. This and the two flanking slices above and below were used for the generation of a maximum intensity projection along the z-axis. The z-projection was blurred with a Gaussian filter (2 pixel diameter). The regions with very bright GFP signals (clones with cells homozygous for the chromosome 3 arm with the GFP marker gene) were manually selected. Similarly, the regions with only background signals in the green channel (clones with cells homozygous for the chromosome 3 arm with ball² and without the GFP marker gene) were also manually delineated, although using a blurred minimum intensity projection of the same three z sections. For each wing disc, we determined the sum of the selected areas for the two types of clones.

Mendaluk A., Caussinus E., Boutros M, & Lehner C.F. (2023). A genome-wide RNAi screen for genes important for proliferation of cultured Drosophila cells at low temperature identifies the Ball/VRK protein kinase. Chromosoma, 132(1), 31-53.

Publication 2023

Cell Nucleus Chromosomes, Human, Pair 3 Clone Cells Cloning Vectors Epistropheus Formaldehyde Genes Genetic Markers Homozygote Imaginal Discs Larva Laser Scanning Microscopy Vision

CUT&RUN protocol for Drosophila wing disc

We performed CUT&RUN in wing disc tissue, as described in (109 (link)), modified from (49 (link)). For each replicate, 20 WL3 imaginal wing discs were used. Two wing discs of Drosophila yakuba were also included with each replicate but were not used in downstream analyses. For the H3K27me3 experiment, αH3K27 (Cell Signaling Technology, no. 9733, 1:100) and protein AG-MNase (1:100; UNC core) were used. For the H3.3 experiment, αH3.3 (H3F33B, 1:100; Abnova, no. H00003021-M01) and protein A-MNase (gift of S. Henikoff) were used.
The ThruPLEX DNA sequencing kit was used for the library preparation. The manufacturer’s protocol was followed until the amplification step. After the addition of indexes, 16 to 21 cycles of 98˚C, 20 s and 67˚C, 10 s were run. DNA library purification was done using AMPure XP beads. Libraries were sequenced on an Illumina NextSeqP2.

Salzler H.R., Vandadi V., McMichael B.D., Brown J.C., Boerma S.A., Leatham-Jensen M.P., Adams K.M., Meers M.P., Simon J.M., Duronio R.J., McKay D.J, & Matera A.G. (2023). Distinct roles for canonical and variant histone H3 lysine-36 in Polycomb silencing. Science Advances, 9(9), eadf2451.

Publication 2023

DNA Library DNA Replication Drosophila Imaginal Discs Proteins Staphylococcal Protein A Tissues

Dissection and Analysis of Drosophila Larval Brains

All flies were reared at 25°C and kept on standard medium. The mutant Tet alleles are described in ^{5 (link),10 (link)}; the wild-type allele used in all experiments is w¹¹¹⁸. Stocks utilized to examine genetic interactions with Tet were sli²/CyO, and robo1⁴/CyO (Bloomington Drosophila Stock center), and robo2^×123/CyO^{26 (link)}. The material used for all whole-genome analysis was either hand dissected third instar larval brains, or, because some experiments necessitated a large input, dissected anterior parts of larvae including the 3 anterior abdominal segments that contain the brain besides other tissues such as imaginal discs, salivary glands, mouth parts and epidermis. Because Tet is highly expressed in the brain and the nerve cell in discs, but not in the other tissues, we call this the Larval Brain Fraction, LBF. Brains and larvae from wt and Tet-GFP third instar larvae were dissected in cold-PBS supplemented with protease inhibitor, snap frozen on dry ice, and stored at −80°C.

Singh B.N., Tran H., Kramer J., Kirishenko E., Changela N., Wang F., Feng Y., Kumar D., Tu M., Lan J., Bizet M., Fuks F, & Steward R. (2023). Tet-dependent 5-hydroxymethyl-Cytosine modification of mRNA regulates the axon guidance genes robo2 and slit in Drosophila. Research Square.

Publication Preprint 2023

Abdomen Alleles Brain Cold Temperature Diptera Drosophila Dry Ice Epidermis Freezing Genome Imaginal Discs Larva Neurons Oral Cavity Protease Inhibitors Salivary Glands Tissues

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What are Imaginal Discs and what is their role in insect development?

Imaginal discs are specialized groups of cells found in the larvae of holometabolous insects, such as flies, beetles, and moths. These discs are the precursors to the adult structures, including wings, legs, antennae, and other external features. During metamorphosis, the imaginal discs undergo rapid growth and differentiation to form the adult insect form. Understanding the development and regulation of imaginal discs is crucial for studying insect morphology, physiology, and evolutionary biology.

What are the different types or variations of Imaginal Discs?

What are some common applications of Imaginal Discs in research?

Imaginal discs are widely studied in various fields of biology, including: 1. Developmental biology: Investigating the mechanisms of growth, patterning, and differentiation of imaginal discs during metamorphosis. 2. Evolutionary biology: Analyzing the morphological changes and adaptations of adult structures derived from imaginal discs across different insect species. 3. Regeneration: Exploring the regenerative potential of imaginal discs and their use in tissue engineering and regenerative medicine.

How can PubCompare.ai assist in optimizing Imaginal Disc research?

PubCompare.ai is an AI-powered platform that can help optimize your imaginal disc research in several ways: 1. It allows you to screen protocol literture more efficiently by leveraging AI to pinpoint critical insights. 2. The platform's AI-driven analysis can highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy. 3. PubCompare.ai can help researchers idenfity the most effective protocols related to Imaginal Discs for their specific research goals, taking their studies to new heights.

More about "Imaginal Discs"

Imaginal discs are the precursory tissue found in the larvae of holometabolous insects, such as flies, beetles, and moths.
These specialized groups of cells are the developmental building blocks that will eventually form the adult structures, including wings, legs, antennae, and other external features.
During the process of metamorphosis, the imaginal discs undergo rapid growth and differentiation, transforming the larval form into the adult insect.
Understanding the development and regulation of imaginal discs is crucial for studying insect morphology, physiology, and evolutionary biology.
Researchers often utilize various tools and techniques to investigate these fascinating structures, including Vectashield mounting medium, Rabbit anti-cleaved caspase-3 antibody, Rabbit anti-GFP antibody, Alexa 488 fluorescent dye, TRIzol for RNA extraction, DAPI for nuclear staining, Cy3 fluorescent dye, and Hoechst 33342 for live-cell imaging.
The Mouse anti-V5 antibody can also be employed to detect and visualize specific proteins within the imaginal discs.
By leveraging the latest advancements in AI-powered platforms, such as PubCompare.ai, researchers can optimize their imaginal disc studies by easily locating the best protocols from literature, pre-prints, and patents, while also benefitting from AI-driven comparisons to enhance reproducibility and accuracy.
This holistic approach can take imaginal disc research to new heights, unlocking a deeper understanding of insect development, physiology, and evolution.