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whole mount in situ hybridization

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https://www.readbyqxmd.com/read/29672592/an-atlas-of-wnt-activity-during-embryogenesis-in-xenopus-tropicalis
#1
Caroline Borday, Karine Parain, Hong Thi Tran, Kris Vleminckx, Muriel Perron, Anne H Monsoro-Burq
Wnt proteins form a family of highly conserved secreted molecules that are critical mediators of cell-cell signaling during embryogenesis. Partial data on Wnt activity in different tissues and at different stages have been reported in frog embryos. Our objective here is to provide a coherent and detailed description of Wnt activity throughout embryo development. Using a transgenic Xenopus tropicalis line carrying a Wnt-responsive reporter sequence, we depict the spatial and temporal dynamics of canonical Wnt activity during embryogenesis...
2018: PloS One
https://www.readbyqxmd.com/read/29660521/an-investigation-of-oyster-tgf-%C3%AE-receptor-genes-and-their-potential-roles-in-early-molluscan-development
#2
Sujian Tan, Pin Huan, Baozhong Liu
Though the roles of BMP signaling development is studied extensively in insects and vertebrates, our knowledge of BMP signaling in molluscan development is limited. In the present study, we performed a genome-based analysis of TGF-β receptors in the Pacific oyster Crassostrea gigas and revealed that C. gigas possessed all five canonical members of the gene family, including three type I and two type II receptors. Whole mount in situ hybridization revealed that four receptor genes exhibited universal expression at the gastrula stage but cgi-bmprII mRNA was only expressed at the anterior lip of blastopore, indicating the regulation of BMP signaling through restricted expression of TGF-β receptors...
April 13, 2018: Gene
https://www.readbyqxmd.com/read/29658397/inflammation-coagulation-response-and-thrombotic-effects-induced-by-silica-nanoparticles-in-zebrafish-embryos
#3
Junchao Duan, Shuang Liang, Yang Yu, Yang Li, Lijing Wang, Zehao Wu, Yueyue Chen, Mark R Miller, Zhiwei Sun
Nowadays, nanotechnology environmental health and safety (nanoEHS) is gaining attention. We previously found that silica nanoparticles (SiNPs) could induce vascular endothelial damage. However, the subsequent toxicologic response to SiNPs-induced endothelial damage was still largely unknown. In this study, we explored the inflammation-coagulation response and thrombotic effects of SiNPs in endothelial cells and zebrafish embryos. For in vitro study, swollen mitochondria and autophagosome were observed in ultrastructural analysis...
April 14, 2018: Nanotoxicology
https://www.readbyqxmd.com/read/29567070/identification-of-duplicated-midkine-genes-and-their-functional-regulation-in-blunt-snout-bream-megalobrama-amblycephala
#4
Dan-Dan Guo, Guo-Dong Zheng, Shang-Ke Du, Bo Qin, Xia-Yun Jiang, Shu-Ming Zou
Midkine (Mdk) is a heparin-binding growth factor that is involved in regulating cell growth, differentiation and migration. Here, we report the isolation and characterization of duplicated mdk genes in blunt snout bream (Megalobrama amblycephala). The mdka and -b genes encode 146 aa and 147 aa peptides, respectively, sharing a sequence identity of 64%. During embryogenesis, mdka mRNA is detectable after 12 h post-fertilization (hpf) and mdkb mRNA can be detected after 8 hpf, about 4 h prior to mdka mRNA...
March 19, 2018: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
https://www.readbyqxmd.com/read/29564768/whole-mount-and-section-in-situ-hybridization-in-mouse-embryos-for-detecting-mrna-expression-and-localization
#5
Kazuko Koshiba-Takeuchi
In situ hybridization is defined as one of the most useful and powerful methods to know where genes (e.g., mRNA, ncRNA) of interest are expressed in tissues. Expression of mRNA can be detected as blue or dark purple signals though hybridization, immunoreaction and coloring steps. Genome-wide approaches in various model animals have been conducted thoroughly, and have led to new research areas aimed at uncovering novel gene functions in cell differentiation and development. To elucidate gene function, spatiotemporal gene expression analysis is very important...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29559927/divergence-of-genes-encoding-cited1-and-cited2-in-blunt-snout-bream-megalobrama-amblycephala-and-their-transcriptional-responses-to-hypoxia
#6
Yuan Sun, Hong-Hong Guo, Dan-Dan Guo, Xia-Yun Jiang, Shu-Ming Zou
The proteins CITED belong to a family of non-DNA-binding transcriptional co-regulators involved in the regulation of various transcriptional responses. Previous studies suggest that members of CITED family may function in response to hypoxia in mammals. however, the molecular and functional information on CITED genes in aquaculture fish is unclear. Here, we characterized and examined the transcriptional patterns of CITED1 and CITED2 genes in the hypoxia-sensitive blunt snout bream ( Megalobrama amblycephala )...
2018: Frontiers in Physiology
https://www.readbyqxmd.com/read/29551495/temporal-and-spatial-expression-of-fgfbp-genes-in-zebrafish
#7
Yana Li, Shuna Sun, Zhiwen Ding, Chunjie Yang, Guoping Zhang, Qiu Jiang, Yunzeng Zou
Fibroblast growth factor binding proteins (FGFBPs) are a class of secreted proteoglycans that function as an extracellular chaperone for locally stored FGFs and enhance FGF signaling. To date, all three human FGFBP genes have been identified and one orthologue fgfbp1a has been studied in zebrafish embryos. Here, we described the cloning and expression patterns of four novel FGFBP orthologues in zebrafish, fgfbp1b, fgfbp2a, fgfbp2b, and fgfbp3. Quantitative PCR and whole-mount in situ hybridization results showed that all transcripts except fgfbp2a are initially expressed in a maternal manner...
March 15, 2018: Gene
https://www.readbyqxmd.com/read/29546721/whole-mount-in-situ-hybridization-and-immunohistochemistry-for-zebrafish-larvae
#8
Rebecca L Cunningham, Kelly R Monk
In situ hybridization enables visualization of mRNA localization, and immunohistochemistry enables visualization of protein localization within a tissue or organism. Both techniques have been extensively utilized in zebrafish (Thisse et al., Development 119:1203-1215, 1993; Dutton et al., Development 128:4113-4125, 2001; Gilmour et al., Neuron 34:577-588, 2002; Lyons et al., Curr Biol 15:513-524, 2005) including for visualization of mRNA localization in Schwann cells (Lyons et al., Curr Biol 15:513-524, 2005; Monk et al...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29496817/whole-mount-in-situ-hybridization-of-xenopus-oocytes
#9
Diana Bauermeister, Tomas Pieler
Whole-mount in situ hybridization (WMISH) is a common approach that is used to visualize spatial and temporal gene expression in embryos. In this process, digoxygenin-labeled antisense RNA is hybridized to the complementary transcript of interest and RNA hybrids are immunohistochemically detected using an alkaline phosphatase-conjugated antibody against digoxigenin. During Xenopus laevis oogenesis, certain RNAs localize to the animal or vegetal pole laying the foundation for germ cell development and germ layer formation of the future embryo...
March 1, 2018: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/29490613/identification-and-characterization-of-micrornas-involved-in-ascidian-larval-metamorphosis
#10
Xiaoming Zhang, Xiaozhuo Liu, Chengzhang Liu, Jiankai Wei, Haiyan Yu, Bo Dong
BACKGROUND: Metamorphosis takes place within the life cycle of most marine invertebrates. The marine ascidian is a classical model to study complex cellular processes and underlying molecular mechanisms involved in its larval metamorphosis. The detailed molecular signaling pathways remain elusive, though extracellular signal-regulated kinases (ERKs) and c-Jun N-terminal kinase (JNK) have been revealed to regulate cell migration, differentiation, and apoptosis in ascidian larval organ regression and juvenile organ development...
March 1, 2018: BMC Genomics
https://www.readbyqxmd.com/read/29486070/expression-and-activity-profiling-of-the-steroidogenic-enzymes-of-glucocorticoid-biosynthesis-and-the-fdx1-co-factors-in-zebrafish
#11
Meltem Weger, Nicolas Diotel, Benjamin D Weger, Tanja Beil, Andreas Zaucker, Helen L Eachus, James A Oakes, Jean Luc do Rego, Karl-Heinz Storbeck, Philipp Gut, Uwe Straehle, Sepand Rastegar, Ferenc Mueller, Nils Krone
The spatial and temporal expression of steroidogenic genes in zebrafish has not been fully characterized. Since zebrafish are increasingly employed in endocrine and stress research, a better characterization of steroidogenic pathways is required to target specific steps in the biosynthetic pathways in the future. Here, we have systematically defined the temporal and spatial expression of steroidogenic enzymes involved in glucocorticoid biosynthesis (cyp21a2, cyp11c1, cyp11a1, cyp11a2, cyp17a1, cyp17a2, hsd3b1, hsd3b2), as well as the mitochondrial electron-providing co-factors ferredoxin (fdx1, fdx1b) during zebrafish development...
February 27, 2018: Journal of Neuroendocrinology
https://www.readbyqxmd.com/read/29462671/expression-of-the-adhesion-g-protein-coupled-receptor-a2-adgra2-during-xenopus-laevis-development
#12
Franziska A Seigfried, Petra Dietmann, Michael Kühl, Susanne J Kühl
The adhesion G protein-coupled receptor A2 (Adgra2) is a seven transmembrane receptor that has been described to be a regulator for angiogenesis in mice. Furthermore, the zebrafish ouchless mutant is unable to develop dorsal root ganglia through a disrupted trafficking of Adgra2. Besides RNA sequencing data, nothing is reported about Adgra2 in the south African crawled frog Xenopus laevis. In this study, we investigated for the first time the spatio-temporal expression of adgra2 during early Xenopus embryogenesis in detail...
February 17, 2018: Gene Expression Patterns: GEP
https://www.readbyqxmd.com/read/29425878/the-klotho-related-protein-klph-lctl-has-preferred-expression-in-lens-and-is-essential-for-expression-of-clic5-and-normal-lens-suture-formation
#13
Jianguo Fan, Joshua Lerner, M Keith Wyatt, Phillip Cai, Katherine Peterson, Lijin Dong, Graeme Wistow
KLPH/lctl belongs to the Klotho family of proteins. Expressed sequence tag analyses unexpectedly revealed that KLPH is highly expressed in the eye lens while northern blots showed that expression is much higher in the eye than in other tissues. In situ hybridization in mouse localized mRNA to the lens, particularly in the equatorial epithelium. Immunofluorescence detected KLPH in lens epithelial cells with highest levels in the germinative/differentiation zone. The gene for KLPH in mouse was deleted by homologous recombination...
February 6, 2018: Experimental Eye Research
https://www.readbyqxmd.com/read/29353190/characterization-of-acrylamide-induced-oxidative-stress-and-cardiovascular-toxicity-in-zebrafish-embryos
#14
Mengmeng Huang, Jingjing Jiao, Jun Wang, Zhidan Xia, Yu Zhang
Acrylamide (AA) is a high production volume chemical in industrial applications and widely found in baked or fried carbohydrate-rich foods. In this study, we unravelled that AA induced developmental toxicity associated with oxidative stress status and disordered lipid distribution in heart region of developing zebrafish. Treatment with AA caused a deficient cardiovascular system with significant heart malformation and dysfunction. We also found that AA could reduce the number of cardiomyocytes through the reduced capacity of cardiomyocyte proliferation rather than cell apoptosis...
April 5, 2018: Journal of Hazardous Materials
https://www.readbyqxmd.com/read/29346450/cloning-and-spatiotemporal-expression-of-xenopus-laevis-apolipoprotein-ci
#15
Jyotsna Sridharan, Tomomi Haremaki, Daniel C Weinstein
Apolipoprotein CI (ApoCI) belongs to the Apolipoprotein superfamily, members of which are involved in lipid transport, uptake and homeostasis. Excessive ApoCI has been implicated in atherosclerosis and Alzheimer's disease in humans. In this study we report the isolation of Xenopus laevis apoCI and describe the expression pattern of this gene during early development, using reverse transcription polymerase chain reaction and whole mount in situ hybridization. Xenopus apoCI is enriched in the dorsal ectoderm during gastrulation, and is subsequently expressed in sensory placodes, neural tube and cranial neural crest...
2018: PloS One
https://www.readbyqxmd.com/read/29322524/a-comprehensive-analysis-of-neurotrophins-and-neurotrophin-tyrosine-kinase-receptors-expression-during-development-of-zebrafish
#16
Valeria Nittoli, Rosa M Sepe, Ugo Coppola, Ylenia D'Agostino, Elena De Felice, Antonio Palladino, Quirino A Vassalli, Annamaria Locascio, Filomena Ristoratore, Antonietta Spagnuolo, Salvatore D'Aniello, Paolo Sordino
Neurotrophins (NTF) are a family of secreted nerve growth factors with affinity for tyrosine kinase (Ntrk) and p75 receptors. To fully understand the variety of developmental roles played by NTFs, it is critical to know when and where genes encoding individual ligands and receptors are transcribed. Identification of ntf and ntrk transcripts in zebrafish development remains to be fully characterized for further uncovering the potential function(s) of the NTF signal transduction pathway. Here, we conducted a systematic analysis of the expression profiles of four ntf and five ntrk genes during zebrafish development using whole-mount in situ hybridization...
April 15, 2018: Journal of Comparative Neurology
https://www.readbyqxmd.com/read/29311262/multidimensional-quantitative-analysis-of-mrna-expression-within-intact-vertebrate-embryos
#17
Vikas Trivedi, Harry M T Choi, Scott E Fraser, Niles A Pierce
For decades, in situ hybridization methods have been essential tools for studies of vertebrate development and disease, as they enable qualitative analyses of mRNA expression in an anatomical context. Quantitative mRNA analyses typically sacrifice the anatomy, relying on embryo microdissection, dissociation, cell sorting and/or homogenization. Here, we eliminate the trade-off between quantitation and anatomical context, using quantitative in situ hybridization chain reaction (qHCR) to perform accurate and precise relative quantitation of mRNA expression with subcellular resolution within whole-mount vertebrate embryos...
January 8, 2018: Development
https://www.readbyqxmd.com/read/29299324/impaired-caudal-fin-fold-regeneration-in-zebrafish-deficient-for-the-tumor-suppressor-pten
#18
Alexander James Hale, Ali Kiai, Jelte Sikkens, Jeroen den Hertog
Zebrafish are able to completely regrow their caudal fin-folds after amputation. Following injury, wound healing occurs, followed by the formation of a blastema, which produces cells to replace the lost tissue in the final phase of regenerative outgrowth. Here we show that, surprisingly, the phosphatase and tumor suppressor Pten, an antagonist of phosphoinositide-3-kinase (PI3K) signaling, is required for zebrafish caudal fin-fold regeneration. We found that homozygous knock-out mutant ( ptena-/- ptenb-/- ) zebrafish embryos, lacking functional Pten, did not regenerate their caudal fin-folds...
August 2017: Regeneration
https://www.readbyqxmd.com/read/29286368/visualizing-multiciliated-cells-in-the-zebrafish-through-a-combined-protocol-of-whole-mount-fluorescent-in-situ-hybridization-and-immunofluorescence
#19
Amanda N Marra, Marisa Ulrich, Audra White, Meghan Springer, Rebecca A Wingert
In recent years, the zebrafish embryo has emerged as a popular model to study developmental biology due to traits such as ex utero embryo development and optical transparency. In particular, the zebrafish embryo has become an important organism to study vertebrate kidney organogenesis as well as multiciliated cell (MCC) development. To visualize MCCs in the embryonic zebrafish kidney, we have developed a combined protocol of whole-mount fluorescent in situ hybridization (FISH) and whole mount immunofluorescence (IF) that enables high resolution imaging...
November 18, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29276941/rnf152-is-essential-for-neurod-expression-and-delta-notch-signaling-in-the-zebrafish-embryos
#20
Ajeet Kumar, Tae-Lin Huh, Joonho Choe, Myungchull Rhee
We report the biological functions of a zebrafish homologue of RING-finger protein 152 (rnf152) during embryogenesis. rnf152 was initially identified as a brain-enriched E3 ligase involved in early embryogenesis of zebrafish. Expression of rnf152 was ubiquitous in the brain at 24 hpf but restricted to the eyes, midbrain-hindbrain boundary (MHB), and rhombomeres at 48 hpf. Knockdown of rnf152 in zebrafish embryos caused defects in the eyes, MHB, and rhombomeres (r1-7) at 24 hpf. These defects in rnf152-deficient embryos were analyzed by whole-mount in situ hybridization (WISH) using neuroD, deltaD, notch1a, and notch3 probes...
December 31, 2017: Molecules and Cells
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