scholarly journals Visualization of Whole-Mount Skeletal Expression Patterns of LacZ Reporters Using a Tissue Clearing Protocol

BioTechniques ◽  
2002 ◽  
Vol 32 (1) ◽  
pp. 66-73 ◽  
Author(s):  
Jitsutaro Kawaguchi ◽  
Valerie Wilson ◽  
Patrick J. Mee
Keyword(s):  
Expression Patterns ◽  
Tissue Clearing ◽  
Whole Mount

mRNA expression patterns of the IGF system during mouse limb bud development, determined by whole mount in situ hybridization

1998 ◽  
Vol 138 (1-2) ◽  
pp. 151-161 ◽  
Author(s):  
Marjolein van Kleffens ◽  
Cora Groffen ◽  
Roberto R. Rosato ◽  
Stefan M. van den Eijnde ◽  
Johan W. van Neck ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Mrna Expression ◽  
Expression Patterns ◽  
Limb Bud ◽  
Bud Development ◽  
Igf System ◽  
Whole Mount ◽  
Mouse Limb

scholarly journals Cytochrome P450 Expression and Chemical Metabolic Activity before Full Liver Development in Zebrafish

2020 ◽  
Vol 13 (12) ◽  
pp. 456
Author(s):  
Tasuku Nawaji ◽  
Natsumi Yamashita ◽  
Haruka Umeda ◽  
Shuangyi Zhang ◽  
Naohiro Mizoguchi ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
In Situ Hybridization ◽  
Metabolic Activity ◽  
Expression Patterns ◽  
Xenobiotic Metabolism ◽  
Liver Development ◽  
P450 Expression ◽  
Whole Mount ◽  
Metabolic Activities

Zebrafish are used widely in biomedical, toxicological, and developmental research, but information on their xenobiotic metabolism is limited. Here, we characterized the expression of 14 xenobiotic cytochrome P450 (CYP) subtypes in whole embryos and larvae of zebrafish (4 to 144 h post-fertilization (hpf)) and the metabolic activities of several representative human CYP substrates. The 14 CYPs showed various changes in expression patterns during development. Many CYP transcripts abruptly increased at about 96 hpf, when the hepatic outgrowth progresses; however, the expression of some cyp1s (1b1, 1c1, 1c2, 1d1) and cyp2r1 peaked at 48 or 72 hpf, before full liver development. Whole-mount in situ hybridization revealed cyp2y3, 2r1, and 3a65 transcripts in larvae at 55 hpf after exposure to rifampicin, phenobarbital, or 2,3,7,8-tetrachlorodibenzo-p-dioxin from 30 hpf onward. Marked conversions of diclofenac to 4′-hydroxydiclofenac and 5-hydroxydiclofenac, and of caffeine to 1,7-dimethylxanthine, were detected as early as 24 or 50 hpf. The rate of metabolism to 4’-hydroxydiclofenac was more marked at 48 and 72 hpf than at 120 hpf, after the liver had become almost fully developed. These findings reveal the expression of various CYPs involved in chemical metabolism in developing zebrafish, even before full liver development.

Expression Patterns of RNAs for Amelin and Amelogenin in Developing Rat Molars and Incisors

1996 ◽  
Vol 10 (2) ◽  
pp. 195-200 ◽  
Author(s):  
C.D. Fong ◽  
L. Hammarström ◽  
C. Lundmark ◽  
T. Wurtz ◽  
I. Slaby
Keyword(s):  
In Situ Hybridization ◽  
Expression Patterns ◽  
Paraffin Sections ◽  
Hybridization Procedure ◽  
Whole Mount ◽  
First Time ◽  
Developing Rat ◽  
Rat Molars ◽  
Oral Epithelia

We have recently identified a novel RNA sequence in ameloblasts, coding for amelin (Cerny et al., 1996). In the present paper, its expression has been compared with that of amelogenin in developing incisors and molars of rats, by means of in situ hybridization of paraffin sections. The RNAs for both amelin and amelogenin were highly expressed in secretory ameloblasts. The expression of RNA for amelogenin gradually decreased in the post-secretory ameloblasts. In contrast, the RNA expression for amelin remained high in post-secretory ameloblasts up to the stage of fusion between dental and oral epithelia at the time of tooth eruption. We suggest that amelin might be involved in the mineralization of enamel or in the attachment of ameloblasts to the enamel surface. The whole-mount in situ hybridization procedure is described for the first time in dental research. It proved to be a useful method and confirmed the results of the conventional in situ hybridization.

scholarly journals All-age whole mount in situ hybridization to reveal larval and juvenile expression patterns in zebrafish

PLoS ONE ◽  
2020 ◽  
Vol 15 (8) ◽  
pp. e0237167
Author(s):  
Franz Vauti ◽  
Luisa A. Stegemann ◽  
Viktoria Vögele ◽  
Reinhard W. Köster
Keyword(s):  
In Situ Hybridization ◽  
Expression Patterns ◽  
Whole Mount

scholarly journals Cellular maps of gastrointestinal organs: getting the most from tissue clearing

2020 ◽  
Vol 319 (1) ◽  
pp. G1-G10
Author(s):  
Cambrian Y. Liu ◽  
D. Brent Polk
Keyword(s):  
Large Scale ◽  
Alimentary Tract ◽  
Three Dimensional ◽  
Short Review ◽  
Biological Tissues ◽  
Tissue Clearing ◽  
Relative Paucity ◽  
Whole Mount ◽  
Chemical Concepts ◽  
The Impact

The development of modern methods to induce optical transparency (“clearing”) in biological tissues has enabled the three-dimensional (3D) reconstruction of intact organs at cellular resolution. New capabilities in visualization of rare cellular events, long-range interactions, and irregular structures will facilitate novel studies in the alimentary tract and gastrointestinal systems. The tubular geometry of the alimentary tract facilitates large-scale cellular reconstruction of cleared tissue without specialized microscopy setups. However, with the rapid pace of development of clearing agents and current relative paucity of research groups in the gastrointestinal field using these techniques, it can be daunting to incorporate tissue clearing into experimental workflows. Here, we give some advice and describe our own experience bringing tissue clearing and whole mount reconstruction into our laboratory’s investigations. We present a brief overview of the chemical concepts that underpin tissue clearing, what sorts of questions whole mount imaging can answer, how to choose a clearing agent, an example of how to clear and image alimentary tissue, and what to do after obtaining the image. This short review will encourage other gastrointestinal researchers to consider how utilizing tissue clearing and creating 3D “maps” of tissue might deepen the impact of their studies.

Heat-Induced Antigen Retrieval Applied in Zebrafish: Whole-Mount In Situ Immunofluorescence Microscopy

2012 ◽  
Vol 18 (3) ◽  
pp. 493-496 ◽  
Author(s):  
Chuang-yu Lin ◽  
Wen-ta Su ◽  
Li-tzu Li
Keyword(s):  
Fluorescent Protein ◽  
Immunofluorescence Microscopy ◽  
Expression Patterns ◽  
Biological Molecules ◽  
Antigen Retrieval ◽  
Zebrafish Embryos ◽  
Fluorescence Dye ◽  
Whole Mount ◽  
Green Fluorescent

AbstractWhole-mount immunofluorescence technique provides a way to reveal integrated expression patterns of biological molecules in individuals. Well-documented morphological preservation ability in biology makes aldehydes the fixative of choice. Cross-linking among biocomponents and aldehydes is the key for maintaining morphology but masks the biological molecules for immunodetection. This study performs an easily accessible method by applying heat-induced retrieval, which can rescue the antigenicity of the proteins and also enhance the labeling sensitivity of the fluorescence dye in overfixed zebrafish embryos. The results show that the immunoreactivities of antibodies to myosin in the muscles, green fluorescent protein in the blood vessels and the nuclei in the cells can be recovered significantly, and the morphology of the zebrafish embryos, even the fragile mutants, is at the same time well maintained. Therefore, we provide a choice for antigen retrieval, which is effective for whole-mount immunofluorescence microscopy.

scholarly journals High-Resolution Whole-MountIn SituHybridization Using Quantum Dot Nanocrystals

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Andriani Ioannou ◽  
Iro Eleftheriou ◽  
Andrea Lubatti ◽  
Anna Charalambous ◽  
Paris A. Skourides
Keyword(s):  
Expression Patterns ◽  
Intracellular Localization ◽  
Emission Spectra ◽  
Fluorescent Imaging ◽  
Proteinase K ◽  
Fluorescent Detection ◽  
Organic Fluorophores ◽  
Whole Mount ◽  
Increased Sensitivity

The photostability and narrow emission spectra of nanometer-scale semiconductor crystallites (QDs) make them desirable candidates for whole-mount fluorescentin situhybridization to detect mRNA transcripts in morphologically preserved intact embryos. We describe a method for direct QD labeling of modified oligonucleotide probes through streptavidin-biotin and antibody-mediated interactions (anti-FITC and anti-digoxigenin). To overcome permeability issues and allow QD conjugate penetration, embryos were treated with proteinase K. The use of QDs dramatically increased sensitivity of whole-mountin situhybridization (WISH) in comparison with organic fluorophores and enabled fluorescent detection of specific transcripts within cells without the use of enzymatic amplification. Therefore, this method offers significant advantages both in terms of sensitivity, as well as resolution. Specifically, the use of QDs alleviates issues of photostability and limited brightness plaguing organic fluorophores and allows fluorescent imaging of cleared embryos. It also offers new imaging possibilities, including intracellular localization of mRNAs, simultaneous multiple-transcript detection, and visualization of mRNA expression patterns in 3D.

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