Read by QxMD icon Read

whole mount in situ hybridization

Iliana Serifi, Eleni Tzima, Katerina Soupsana, Zoe Karetsou, Dimitris Beis, Thomais Papamarcaki
The oncoprotein SET/I2PP2A participates in various cellular mechanisms such as transcription, cell cycle regulation and cell migration. SET is also an inhibitor of the serine/threonine phosphatase PP2A, which is involved in the regulation of cell homeostasis. In zebrafish there are two paralogous set genes that encode Seta (269 aa) and Setb (275 aa) proteins which share 94% identity. We show here that seta and set b are similarly expressed in the eye, the otic vesicle, the brain and the lateral line system, as indicated by in situ hybridization labeling...
October 17, 2016: Biochemical Journal
Tomoichiro Yamaai, Masaharu Takigawa
A procedure for whole-mount in situ hybridization developed for detecting gene expression of Ccn2 in developing calcified tissues of mouse embryos is presented. In this method, embryos are hybridized with Dig-labeled riboprobes, and the riboprobes are detected by use of the alkaline-phosphatase reaction in the presence of a 4-nitro-blue tetrazolium chloride and 5-bromo-4-chloro-3-indolyl-phosphate (NBT + BCIP) mixture. Obvious detection of positive signals for Ccn2 in the cartilage of developing phalanges indicates that this method can be applied to gene expression analysis of other Ccn genes in developing calcified tissues...
2017: Methods in Molecular Biology
Bruce W Draper
Both male and female zebrafish have a population of germ-line stem cells that produce gametes throughout the life of the fish. These cells localize to specific regions in the gonads and can be identified because they uniquely express the nanos2 gene, which encodes a conserved regulator of translation. A method is presented here for identifying germ-line stem cells in the ovary and testis using a combined protocol for whole-mount fluorescent RNA in situ hybridization to detect nanos2 mRNA and immunofluorescence to detect the pan-germ cell marker Vasa...
2017: Methods in Molecular Biology
Julie N R Collins, James J Collins
Schistosomes are flatworm parasites that claim the lives of more than 200,000 people in poverty-stricken regions every year. Much of the pathology due to infection is the direct result of injury spurred by the parasite's eggs becoming lodged in host tissues. Thus, asking basic questions about germ cell biology may not only identify novel therapeutic approaches, but could also uncover conserved mechanisms that regulate the germline in diverse metazoa. Here, we detail useful methods for studying the schistosome germline including EdU labeling, whole-mount in situ hybridization, and RNA interference...
2017: Methods in Molecular Biology
Wim Jonckheere, Wannes Dermauw, Vladimir Zhurov, Nicky Wybouw, Jan Van den Bulcke, Carlos A Villarroel, Robert Greenhalgh, Mike Grbić, Rob C Schuurink, Luc Tirry, Geert Baggerman, Richard M Clark, Merijn R Kant, Bartel Vanholme, Gerben Menschaert, Thomas Van Leeuwen
The two-spotted spider mite Tetranychus urticae is an extremely polyphagous crop pest. Alongside an unparalleled detoxification potential for plant secondary metabolites, it has recently been shown that spider mites can attenuate or even suppress plant defenses. Salivary constituents, notably effectors, have been proposed to play an important role in manipulating plant defenses and might determine the outcome of plant-mite interactions. Here, the proteomic composition of saliva from T. urticae lines adapted to various host plants - bean, maize, soy, and tomato - was analyzed using a custom-developed feeding assay coupled with nano-LC tandem mass spectrometry...
October 4, 2016: Molecular & Cellular Proteomics: MCP
Harry M T Choi, Colby R Calvert, Naeem Husain, David Huss, Julius C Barsi, Benjamin E Deverman, Ryan C Hunter, Mihoko Kato, S Melanie Lee, Anna C T Abelin, Adam Z Rosenthal, Omar S Akbari, Yuwei Li, Bruce A Hay, Paul W Sternberg, Paul H Patterson, Eric H Davidson, Sarkis K Mazmanian, David A Prober, Matt van de Rijn, Jared R Leadbetter, Dianne K Newman, Carol Readhead, Marianne E Bronner, Barbara Wold, Rusty Lansford, Tatjana Sauka-Spengler, Scott E Fraser, Niles A Pierce
In situ hybridization methods are used across the biological sciences to map mRNA expression within intact specimens. Multiplexed experiments, in which multiple target mRNAs are mapped in a single sample, are essential for studying regulatory interactions, but remain cumbersome in most model organisms. Programmable in situ amplifiers based on the mechanism of hybridization chain reaction (HCR) overcome this longstanding challenge by operating independently within a sample, enabling multiplexed experiments to be performed with an experimental timeline independent of the number of target mRNAs...
October 1, 2016: Development
Shuang Jiao, Zhihao Wu, Xungang Tan, Yulei Sui, Lijuan Wang, Feng You
Although chromosome set manipulation techniques including polyploidy induction and gynogentic induction in flatfish are becoming increasingly mature, there exists a poor understanding of their effects on embryonic development. PAX3 plays crucial roles during embryonic myogenesis and neurogenesis. In olive flounder (Paralichthys olivaceus), there are two duplicated pax3 genes (pax3a, pax3b), and both of them are expressed in the brain and muscle regions with some subtle regional differences. We utilized pax3a and pax3b as indicators to preliminarily investigate whether chromosome set manipulation affects embryonic neurogenesis and myogenesis using whole-mount in situ hybridization...
September 27, 2016: Fish Physiology and Biochemistry
Li Yao, Shi-Xiao Peng, Yi-Da Xu, Stanley Li Lin, Yu-Hong Li, Chun-Jie Liu, Hou-De Zhao, Lin-Fang Wang, Yan-Qin Shen
1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP), which induces the pathological characteristics of Parkinson's disease in rodents, also specifically targets dopaminergic neurons in zebrafish embryos and larvae. Loganin, a traditional Chinese drug, was reported to regulate immune function and possess anti-inflammatory and anti-shock effects. Here, we investigate the role of loganin in MPTP-induced Parkinson-like abnormalities in zebrafish. MPTP treatment-induced abnormal development, in larvae, such as pericardium edema, increased yolk color, yolk sac edema, and retarded yolk sac resorption, as well as defects in brain development...
September 23, 2016: Journal of Cellular Biochemistry
Li-Chuan Tseng, Chih-Hao Tang, Yun-Jin Jiang
High-throughput screening with a loss-of-function strategy is a logical and efficient way to identify novel genes involved in biological processes of interest. In zebrafish, morpholinos have been developed as a convenient tool to knock down gene expression. Here, we describe procedures for systematic screening using morpholinos in zebrafish to identify novel deubiquitylases involved in convergent extension during gastrulation. In this example, we examine candidates based on embryonic morphology and molecular signals of whole mount in situ hybridization assay...
2016: Methods in Molecular Biology
Yuansi Zheng, Jiebo Lin, Jieyi Li, Haifeng Zhang, Weiming Ai, Xuedong Wang, Randy A Dahlgren, Huili Wang
The toxicity of β-diketone antibiotics (DKAs), a class of ''pseudo-persistent'' environmental pollutants, to F0-zebrafish (Danio rerio) was investigated using 7-dpf F1-zebrafish miRNA sequencing and bioinformatics analyses. Based on relative expression, 47, 134 and 118 of 193 mature miRNAs were differentially expressed between control vs 6.25 mg/L, control vs 12.5 mg/L and 6.25 vs 12.5 mg/L treatments, respectively. Utilizing three databases, 2523 potential target genes were predicted, and they were assigned to 19 high-abundance KEGG pathways and 20 functional categories by COG analysis...
December 2016: Chemosphere
Troy Camarata, Aleksandr Vasilyev, Michael Hadjiargyrou
Mustn1 is a small nuclear protein that is involved in the development and regeneration of the musculoskeletal system. Previous work established a role for Mustn1 in myogenic and chondrogenic differentiation. In addition, recent evidence suggests a potential role for Mustn1 in cilia function in zebrafish. A detailed study of Mustn1 expression has yet to be conducted in zebrafish. As such, we report herein the cloning of the zebrafish Mustn1 orthologs, mustn1a and mustn1b, and their expression during zebrafish embryonic and larval development...
November 15, 2016: Gene
Joanne B Cole, Mange Manyama, Emmanuel Kimwaga, Joshua Mathayo, Jacinda R Larson, Denise K Liberton, Ken Lukowiak, Tracey M Ferrara, Sheri L Riccardi, Mao Li, Washington Mio, Michaela Prochazkova, Trevor Williams, Hong Li, Kenneth L Jones, Ophir D Klein, Stephanie A Santorico, Benedikt Hallgrimsson, Richard A Spritz
The human face is a complex assemblage of highly variable yet clearly heritable anatomic structures that together make each of us unique, distinguishable, and recognizable. Relatively little is known about the genetic underpinnings of normal human facial variation. To address this, we carried out a large genomewide association study and two independent replication studies of Bantu African children and adolescents from Mwanza, Tanzania, a region that is both genetically and environmentally relatively homogeneous...
August 2016: PLoS Genetics
Pezhman Salehi, Anthony Myint, Young J Kim, Marshall X Ge, Joel Lavinsky, Maria K Ho, Amanda L Crow, Charlene Cruz, Maya Monges-Hernadez, Juemei Wang, Jaana Hartiala, Li I Zhang, Hooman Allayee, Aldons J Lusis, Takahiro Ohyama, Rick A Friedman
This study aimed to investigate the genetic causes of vestibular dysfunction. We used vestibular sensory-evoked potentials (VsEPs) to characterize the vestibular function of 35 inbred mouse strains selected from the Hybrid Mouse Diversity Panel and demonstrated strain-dependent phenotypic variation in vestibular function. Using these phenotypic data, we performed the first genome-wide association study controlling for population structure that has revealed two highly suggestive loci, one of which lies within a haplotype block containing five genes (Stard6, 4930503L19Rik, Poli, Mbd2, Dcc) on Chr...
October 2016: Journal of the Association for Research in Otolaryngology: JARO
Louise S Goupil, Sam L Ivry, Ivy Hsieh, Brian M Suzuki, Charles S Craik, Anthony J O'Donoghue, James H McKerrow
Proteases perform numerous vital functions in flatworms, many of which are likely to be conserved throughout the phylum Platyhelminthes. Within this phylum are several parasitic worms that are often poorly characterized due to their complex life-cycles and lack of responsiveness to genetic manipulation. The flatworm Schmidtea mediterranea, or planaria, is an ideal model organism to study the complex role of protein digestion due to its simple life cycle and amenability to techniques like RNA interference (RNAi)...
August 2016: PLoS Neglected Tropical Diseases
Yongan Wang, Qing Yang, Wei Liu, Mingxi Yu, Zhou Zhang, Xiaoyu Cui
Human's ubiquitous exposure to di (2-ethylhexyl) phthalate (DEHP) is thought to be associated with female reproductive toxicity. Previous studies found that DEHP inhibited follicle growth and decreased estradiol levels in adult female mice. However, limited information is available on the link between in utero DEHP exposure and ovarian development in female mouse offspring. The present study evaluates the disturbances in regulatory genes involved in female sex determination and the ovarian outcomes in fetal and postnatal female mice treated with in utero DEHP exposure...
September 15, 2016: Toxicology and Applied Pharmacology
Giovanni Anthony, Ju-Ahng Lee
Recent development in 3D printing technology has opened an exciting possibility for manufacturing 3D devices on one's desktop. We used 3D modeling programs to design 3D models of a tissue-handling system and these models were "printed" in a stereolithography (SLA) 3D printer to create precision histology devices that are particularly useful to handle multiple samples with small dimensions in parallel. Our system has been successfully tested for in situ hybridization of zebrafish embryos. Some of the notable features include: (1) A conveniently transferrable chamber with 6 mesh-bottomed wells, each of which can hold dozens of zebrafish embryos...
2016: PloS One
Shi-Zhu Li, Wei Liu, Zhi Li, Yang Wang, Li Zhou, Mei-Sheng Yi, Jian-Fang Gui
As a germ cell marker gene, Dead end (dnd) has been identified and characterized in many vertebrates. Recently, we created a complete germ cell-depleted gonad model by the dnd-specific morpholino-mediated knockdown approach, and revealed sex-biased gene expression alteration through utilizing unisexual gynogenetic superiority in polyploid gibel carp. However, dnd and its expression pattern are still unclear in the gibel carp. In this study, we further analyzed molecular characterization of gibel carp dnd and its dynamic expression pattern during gametogenesis and embryogenesis...
October 10, 2016: Gene
Mouli Ghosh Dastidar, Magdalena Mosiolek, Andrea Bleckmann, Thomas Dresselhaus, Michael D Nodine, Alexis Maizel
Small RNAs such as microRNAs (miRNAs), regulate gene expression and play important roles in many plant processes. Although our knowledge of their biogenesis and mode of action has significantly progressed, we comparatively still know little about their biological functions. In particular, knowledge about their spatiotemporal expression patterns rely on either indirect detection by use of reporter constructs or labor-intensive direct detection by in situ hybridization on sectioned material. None of the current approaches allows a systematic investigation of small RNA expression patterns...
July 14, 2016: Plant Journal: for Cell and Molecular Biology
Soo-Ho Lee, Hyun-Kyung Lee, Chowon Kim, Yoo-Kyung Kim, Tayaba Ismail, Youngeun Jeong, Kyeongyeon Park, Jeen-Woo Park, Oh-Shin Kwon, Beom Sik Kang, Dong-Seok Lee, Mae-Ja Park, Do-Sim Park, Hyun-Shik Lee
Alternative splicing is a major mechanism regulating pattern of gene expression through the production of multiple mRNAs from a single gene transcript. Any misregulation can cause various human diseases and also have severe effects on embryogenesis. SRSF1 is one of the critical factors regulating alternative splicing at many stages of vertebrate development and any disturbance in SRSF1 leads to serious consequences. In current study, we investigated the effects of loss of the SRSF1 gene using antisense morpholino oligonucleotides (MO) in Xenopus embryogenesis...
September 2, 2016: Biochemical and Biophysical Research Communications
Nooreen Reza, Mustafa K Khokha, Florencia Del Viso
Nucleoporins (nups) compose the structure of the nuclear pore complex (NPC) of all cells, but several studies have illuminated nucleoporins' additional roles in development and the cell cycle. However, a comprehensive study of nup expression in embryonic development has not yet been reported. We synthesized antisense probes for all nup genes and used whole-mount in situ hybridization techniques to determine the expression pattern of all members of the nup family of genes at three different developmental stages in Xenopus tropicalis...
2016: International Journal of Developmental Biology
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"