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whole mount in situ hybridization

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https://www.readbyqxmd.com/read/28427488/nr2e1-downregulation-is-involved-in-excess-retinoic-acid-induced-developmental-abnormality-in-the-mouse-brain
#1
Juan Yu, Qian Guo, Jian Bing Mu, Ting Zhang, Ren Ke Li, Jun Xie
OBJECTIVE: This study aimed to investigate the expression pattern and function of Nuclear receptor subfamily 2 group E member 1 (Nr2e1) in retinoic acid (RA)-induced brain abnormality. METHODS: The mouse model of brain abnormality was established by administering 28 mg/kg RA, and neural stem cells (NSCs) were isolated from the mouse embryo and cultured in vitro. Nr2e1 expression was detected by whole mount in situ hybridization, RT-PCR, and Western blotting. Nr2e1 function was determined by transducing Nr2e1 shRNA into NSCs, and the effect on the sonic hedgehog (Shh) signaling pathway was assessed in the cells...
March 2017: Biomedical and Environmental Sciences: BES
https://www.readbyqxmd.com/read/28414113/sept8a-and-sept8b-mrna-expression-in-the-developing-and-adult-zebrafish
#2
Constantin Berger, Frederik Helmprobst, Prisca Chapouton, Christina Lillesaar, Christian Stigloher
Septins are highly conserved GTP-binding proteins involved in numerous cellular processes. Despite a growing awareness of their roles in the cell biology, development and signal transmission in nervous systems, comparably little is known about precise septin expression. Here, we use the well-established model organism zebrafish (Danio rerio) to unravel the expression of sept8a and sept8b, with special focus on the CNS. We performed whole mount RNA in situ hybridization on zebrafish 1-4 dpf in combination with serial sectioning of epon-embedded samples as well as on brain sections of adult zebrafish to obtain precise histological mapping of gene expression...
April 14, 2017: Gene Expression Patterns: GEP
https://www.readbyqxmd.com/read/28392476/schistosoma-mansoni-displays-an-adenine-phosphoribosyltransferase-preferentially-expressed-in-mature-female-gonads-and-vitelaria
#3
Ana Eliza Zeraik, Vitor Hugo Balasco Serrão, Larissa Romanello, Juliana Roberta Torini, Alexandre Cassago, Ricardo DeMarco, Humberto D'Muniz Pereira
Schistosoma mansoni depends upon the purine salvage pathway to obtain purine nucleotides; therefore, enzymes from this pathway are essential for parasite survival. Here, we focused on the adenine phosphoribosyltransferase (APRT) enzyme, which catalyzes the condensation reaction between adenine and PRPP (5-phosphoribosylpyrophosphate) to produce AMP and PPi. Kinetic experiments using the heterologously expressed protein of one APRT isoform from S. mansoni indicate that it is catalytically active, and whole-mount in situ hybridization studies indicate that the transcripts of this protein are concentrated in the posterior region of the ovary and vitellaria of female adult worms...
April 6, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28344782/la-related-protein-6-controls-ciliated-cell-differentiation
#4
Zarko Manojlovic, Ryan Earwood, Akiko Kato, Diana Perez, Oscar A Cabrera, Ruth Didier, Timothy L Megraw, Branko Stefanovic, Yoichi Kato
BACKGROUND: La-related protein 6 (LARP6) is an evolutionally conserved RNA-binding protein. Vertebrate LARP6 binds the 5' stem-loop found in mRNAs encoding type I collagen to regulate their translation, but other target mRNAs and additional functions for LARP6 are unknown. The aim of this study was to elucidate an additional function of LARP6 and to evaluate the importance of its function during development. METHODS: To uncover the role of LARP6 in development, we utilized Morpholino Oligos to deplete LARP6 protein in Xenopus embryos...
2017: Cilia
https://www.readbyqxmd.com/read/28324191/planarian-homolog-of-puromycin-sensitive-aminopeptidase-djpsa-is-required-for-brain-regeneration
#5
Suge Wu, Bin Liu, Zuoqing Yuan, Xiufang Zhang, Hong Liu, Qiuxiang Pang, Bosheng Zhao
Puromycin-sensitive aminopeptidase (PSA) belongs to the M1 zinc metallopeptidase family. PSA is the most abundant aminopeptidase in the brain and plays a role in the metabolism of neuropeptides including those involved in neurodegeneration. A cDNA DjPsa was identified from the planarian Dugesia japonica cDNA library. It contains a 639-bp open reading frame corresponding to a deduced protein of 212 amino acids. Whole mount in situ hybridization revealed that DjPsa is expressed in the brain and ventral nerve cords of intact and regenerating animals and demonstrates a tissue and stage-specific expression pattern of DjPsa in developing embryos and larvae...
June 2017: Invertebrate Neuroscience: IN
https://www.readbyqxmd.com/read/28280925/a-soxc-gene-related-to-larval-shell-development-and-co-expression-analysis-of-different-shell-formation-genes-in-early-larvae-of-oyster
#6
Gang Liu, Pin Huan, Baozhong Liu
Among the potential larval shell formation genes in mollusks, most are expressed in cells surrounding the shell field during the early phase of shell formation. The only exception (cgi-tyr1) is expressed in the whole larval mantle and thus represents a novel type of expression pattern. This study reports another gene with such an expression pattern. The gene encoded a SoxC homolog of the Pacific oyster Crassostrea gigas and was named cgi-soxc. Whole-mount in situ hybridization revealed that the gene was highly expressed in the whole larval mantle of early larvae...
March 9, 2017: Development Genes and Evolution
https://www.readbyqxmd.com/read/28256719/rna-whole-mount-in-situ-hybridization-proximity-ligation-assay-rish-pla-an-assay-for-detecting-rna-protein-complexes-in-intact-cells
#7
Ioannis M Roussis, Fiona A Myers, Garry P Scarlett
Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hybridization-Proximity Ligation Assay (ISH-PLA) to the study of RNA regulation (rISH-PLA). The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single-cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogeneous cell populations from which conventional RNA-protein interaction detection techniques suffer...
March 3, 2017: Current Protocols in Cell Biology
https://www.readbyqxmd.com/read/28228749/neuroprotective-role-of-the-pi3-kinase-akt-signaling-pathway-in-zebrafish
#8
Shuang Chen, Yunzhang Liu, Xiaozhi Rong, Yun Li, Jianfeng Zhou, Ling Lu
Neuronal survival and growth in the embryo is controlled partly by trophic factors. For most trophic factors (such as Insulin-like growth factor-1), the ability to regulate cell survival has been attributed to the phosphoinositide 3-kinase (PI3K)/Akt kinase cascade. This study presents data illustrating the role of PI3K/Akt in attainment of normal brain size during zebrafish embryogenesis. Blocking PI3K with inhibitor LY294002 caused a significant reduction in brain size (in addition to global growth retardation) during zebrafish embryogenesis...
2017: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/28217803/-natriuretic-peptide-precursor-c-coding-gene-contributes-to-zebrafish-angiogenesis
#9
Jing-Jing Zhang, Xin Wang, Dong Liu
This study aimed to investigate the expression of the natriuretic peptide precursor C coding gene nppc and its role in angiogenesis during embryonic period of the zebrafish. Whole mount in situ hybridization was performed to detect the expression pattern of nppc. nppc specific morpholino and nppc mRNA were injected respectively into the one-cell stage embryo to specifically knock-down and rescue the expression of nppc in Tg (flk1:GFP) and Tg (fli1a:nGFP) transgenic lines. The morphology and endothelial cell number of intersegmental vessel (ISV) were analyzed after imaging using the laser scanning confocal microscope...
February 25, 2017: Sheng Li Xue Bao: [Acta Physiologica Sinica]
https://www.readbyqxmd.com/read/28213042/gene-duplication-conservation-and-divergence-of-heme-oxygenase-2-genes-in-blunt-snout-bream-megalobrama-amblycephala-and-their-responses-to-hypoxia
#10
Xue-Li Zhang, Yi-Wen Sun, Jie Chen, Xia-Yun Jiang, Shu-Ming Zou
Heme oxygenase (HO) that catalyzes the degradation of heme, is involved in responding and using oxygen in teleost fish. Physiologic heme degradation can be catalyzed by two isozymes of HO (HO-1 and HO-2). In fish, the molecular constructions, expression characteristics and hypoxic regulation of HO-2 are still not well known. Here, we report the isolation and characterization of duplicated HO-2 genes in blunt snout bream, a hypoxia sensitive fish species. Blunt snout bream HO-2a and -2b genes shared a relatively low sequence identity of 67%...
April 30, 2017: Gene
https://www.readbyqxmd.com/read/28197779/colonic-mesenchyme-differentiates-into-smooth-muscle-before-its-colonization-by-vagal-enteric-neural-crest-derived-cells-in-the-chick-embryo
#11
Annick Bourret, Norbert Chauvet, Pascal de Santa Barbara, Sandrine Faure
During development, the gastrointestinal (GI) tract arises from a primary tube composed of mesoderm and endoderm. The mesoderm gives rise to the digestive mesenchyme, which in turn differentiates into multiple tissues, namely the submucosa, the interstitial cells of Cajal and the smooth muscle cells (SMCs). Concomitant with these early patterning events, the primitive GI tract is colonized by vagal enteric neural crest-derived cells (vENCDCs), a population of cells that gives rise to the enteric nervous system, the intrinsic innervation of the GI tract...
February 15, 2017: Cell and Tissue Research
https://www.readbyqxmd.com/read/28158974/a-compendium-of-developmental-gene-expression-in-lake-malawi-cichlid-fishes
#12
R F Bloomquist, T E Fowler, J B Sylvester, R J Miro, J T Streelman
BACKGROUND: Lake Malawi cichlids represent one of a growing number of vertebrate models used to uncover the genetic and developmental basis of trait diversity. Rapid evolutionary radiation has resulted in species that share similar genomes but differ markedly in phenotypes including brains and behavior, nuptial coloration and the craniofacial skeleton. Research has begun to identify the genes, as well as the molecular and developmental pathways that underlie trait divergence. RESULTS: We assemble a compendium of gene expression for Lake Malawi cichlids, across pharyngula (the phylotypic stage) and larval stages of development, encompassing hundreds of gene transcripts...
February 3, 2017: BMC Developmental Biology
https://www.readbyqxmd.com/read/28118669/the-ciliopathy-gene-ahi1-is-required-for-zebrafish-cone-photoreceptor-outer-segment-morphogenesis-and-survival
#13
Emma M Lessieur, Joseph Fogerty, Robert J Gaivin, Ping Song, Brian D Perkins
Purpose: Joubert syndrome (JBTS) is an autosomal recessive ciliopathy with considerable phenotypic variability. In addition to central nervous system abnormalities, a subset of JBTS patients exhibit retinal dystrophy and/or kidney disease. Mutations in the AHI1 gene are causative for approximately 10% of all JBTS cases. The purpose of this study was to generate ahi1 mutant alleles in zebrafish and to characterize the retinal phenotypes. Methods: Zebrafish ahi1 mutants were generated using transcription activator-like effector nucleases (TALENs)...
January 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28104549/mechanisms-of-silver_nanoparticles-induced-hypopigmentation-in-embryonic-zebrafish
#14
Lian Xu, Qin-Han Xu, Xin-Ying Zhou, Li-Yan Yin, Peng-Peng Guan, Ting Zhang, Jing-Xia Liu
Silver_nanoparticles (AgNPs) have been reported to inhibit specification of erythroid cells and to induce spinal cord deformities and cardiac arrhythmia in vertebrates, but have not been implicated in development of neural crest (NC) and pigment cells in an in vivo model yet. In current study, down-regulated expressions of NC genes pax7 and foxd3, melanophore genes mitfa and dct, and xanthophore gene gch2 in AgNPs-exposed embryos were revealed by microarray, qRT-PCR and whole-mount in situ hybridization (WISH)...
March 2017: Aquatic Toxicology
https://www.readbyqxmd.com/read/28096217/high-throughput-analysis-reveals-novel-maternal-germline-rnas-crucial-for-primordial-germ-cell-preservation-and-proper-migration
#15
Dawn A Owens, Amanda M Butler, Tristan H Aguero, Karen M Newman, Derek Van Booven, Mary Lou King
During oogenesis, hundreds of maternal RNAs are selectively localized to the animal or vegetal pole, including determinants of somatic and germline fates. Although microarray analysis has identified localized determinants, it is not comprehensive and is limited to known transcripts. Here, we utilized high-throughput RNA-sequencing analysis to comprehensively interrogate animal and vegetal pole RNAs in the fully grown Xenopus laevis oocyte. We identified 411 (198 annotated) and 27 (15 annotated) enriched mRNAs at the vegetal and animal pole, respectively...
January 15, 2017: Development
https://www.readbyqxmd.com/read/28078730/a-fern-aintegumenta-gene-mirrors-baby-boom-in-promoting-apogamy-in-ceratopteris-richardii
#16
Linh T Bui, Dzevida Pandzic, Christopher E Youngstrom, Simon Wallace, Erin E Irish, Péter Szövényi, Chi-Lien Cheng
Asexual reproduction is widespread in land plants, including ferns where 10% of all species are obligate asexuals. In these ferns, apogamous sporophytes are generated directly from gametophytes, bypassing fertilization. In the model fern Ceratopteris richardii, a sexual species, apogamy can be induced by culture on high sugar media. BABY BOOM (BBM) genes in angiosperms are known to promote somatic embryogenesis, which like apogamy produce sporophytes without fertilization. Here, a Brassica napus BBM (BnBBM) was used to investigate genetic similarity between apogamy in ferns and somatic embryogenesis in angiosperms...
January 12, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28048959/expression-and-localization-of-carbonic-anhydrase-genes-in-the-serpulid-polychaete-hydroides-elegans
#17
Grant Batzel, Brian T Nedved, Michael G Hadfield
The metalloenzyme, carbonic anhydrase (CA), catalyzes the reversible hydration of carbon dioxide into bicarbonate, and is responsible for biomineralization processes in animals. In the Annelida, the marine worms in the family Serpulidae are typified by the construction of calcium carbonate tubes. Hydroides elegans, a common member of warmwater biofouling communities around the world, provides an outstanding model for studies of calcification. To better understand the molecular process of biomineralization in H...
December 2016: Biological Bulletin
https://www.readbyqxmd.com/read/28034797/expression-profile-of-rrbp1-genes-during-embryonic-development-and-in-adult-tissues-of-xenopus-laevis
#18
Guang-Hui Liu, Cheng-Zhou Mao, Hai-Yan Wu, Deng-Cheng Zhou, Jing-Bo Xia, Soo-Ki Kim, Dong-Qing Cai, Hui Zhao, Xu-Feng Qi
Recent studies suggest that ribosome-binding protein 1 (RRBP1) is involved in multiple diseases such as tumorigenesis and cardiomyopathies. However, its function during embryonic development remains largely unknown. We searched Xenopus laevis database with human RRBP1 protein sequence and identified two cDNA sequences encoding Xenopus orthologs of RRBP1 including rrbp1a (NM_001089623) and rrbp1b (NM_001092468). Both genes were firstly detected at blastula stage 8 with weak signals in animal hemisphere by whole mount in situ hybridization...
January 2017: Gene Expression Patterns: GEP
https://www.readbyqxmd.com/read/28013423/cloning-expression-profiling-and-promoter-functional-analysis-of-bone-morphogenetic-protein-6-and-7-in-tongue-sole-cynoglossus-semilaevis
#19
Qian Ma, Wenrong Feng, Zhimeng Zhuang, Shufang Liu
Bone morphogenetic proteins (BMPs) play crucial roles in vertebrate developmental process and are associated with the mechanisms which drive early skeletal development. As a first approach to elucidating the role of BMPs in regulating fish bone formation and growth, we describe the cloning, expression profiling and promoter functional analysis of bmp6 and bmp7 in tongue sole (Cynoglossus semilaevis). The full length of bmp6 and bmp7 cDNA sequences is 1939 and 1836 bp, which encodes a protein of 428 and 427 amino acids, respectively...
December 24, 2016: Fish Physiology and Biochemistry
https://www.readbyqxmd.com/read/28002765/developmental-toxicity-of-2-4-dichlorophenoxyacetic-acid-in-zebrafish-embryos
#20
Kang Li, Jia-Qi Wu, Ling-Ling Jiang, Li-Zhen Shen, Jian-Ying Li, Zhi-Heng He, Ping Wei, Zhuo Lv, Ming-Fang He
2,4-Dichlorophenoxyacetic acid (2,4-D) is widely used in agriculture as herbicide/pesticide, plant growth regulator and fruit preservative agent. It progressively accumulates in the environment including surface water, air and soil. It could be detected in human food and urine, which poses great risk to the living organisms. In the present study, we investigated the developmental toxicity of 2,4-D on zebrafish (Danio rerio) embryo. 2,4-D exposure significantly decreased both the survival rate (LC50 = 46...
December 10, 2016: Chemosphere
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