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whole mount in situ hybridization

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https://www.readbyqxmd.com/read/28096217/high-throughput-analysis-reveals-novel-maternal-germline-rnas-crucial-for-primordial-germ-cell-preservation-and-proper-migration
#1
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
#2
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
#3
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
#4
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...
December 26, 2016: 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
#5
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
#6
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
https://www.readbyqxmd.com/read/27990106/localization-of-neuropeptide-gene-expression-in-larvae-of-an-echinoderm-the-starfish-asterias-rubens
#7
Tatiana D Mayorova, Shi Tian, Weigang Cai, Dean C Semmens, Esther A Odekunle, Meet Zandawala, Yusef Badi, Matthew L Rowe, Michaela Egertová, Maurice R Elphick
Neuropeptides are an ancient class of neuronal signaling molecules that regulate a variety of physiological and behavioral processes in animals. The life cycle of many animals includes a larval stage(s) that precedes metamorphic transition to a reproductively active adult stage but, with the exception of Drosophila melanogaster and other insects, research on neuropeptide signaling has hitherto largely focused on adult animals. However, recent advances in genome/transcriptome sequencing have facilitated investigation of neuropeptide expression/function in the larvae of protostomian (e...
2016: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/27987051/expression-patterns-indicate-that-bmp2-4-and-chordin-not-bmp5-8-and-gremlin-mediate-dorsal-ventral-patterning-in-the-mollusk-crassostrea-gigas
#8
Sujian Tan, Pin Huan, Baozhong Liu
Though several bilaterian animals use a conserved BMP2/4-Chordin antagonism to pattern the dorsal-ventral (DV) axis, the only lophotrochozoan species in which early DV patterning has been studied to date, the leech Helobdella robusta, appears to employ BMP5-8 and Gremlin. These findings call into question the conservation of a common DV patterning mechanism among bilaterian animals. To explore whether the unusual DV patterning mechanism in H. robusta is also used in other lophotrochozoan species, we investigated the expression of orthologous genes in the early embryo of a bivalve mollusk, Crassostrea gigas...
December 16, 2016: Development Genes and Evolution
https://www.readbyqxmd.com/read/27936201/sterol-o-acyltransferase-2-contributes-to-the-yolk-cholesterol-trafficking-during-zebrafish-embryogenesis
#9
Nai-Yun Chang, Yen-Ju Chan, Shih-Torng Ding, Yen-Hua Lee, Wei-Chun HuangFu, I-Hsuan Liu
To elucidate whether Sterol O-acyltransferase (Soat) mediates the absorption and transportation of yolk lipids to the developing embryo, zebrafish soat1 and soat2 were cloned and studied. In the adult zebrafish, soat1 was detected ubiquitously while soat2 mRNA was detected specifically in the liver, intestine, brain and testis. Whole mount in situ hybridization demonstrated that both soat1 and soat2 expressed in the yolk syncytial layer, hatching gland and developing cardiovascular as well as digestive systems, suggesting that Soats may play important roles in the lipid trafficking and utilization during embryonic development...
2016: PloS One
https://www.readbyqxmd.com/read/27895889/amino-acid-transporters-implicated-in-endocytosis-of-buchnera-during-symbiont-transmission-in-the-pea-aphid
#10
Hsiao-Ling Lu, Chun-Che Chang, Alex C C Wilson
BACKGROUND: Many insects host their obligate, maternally transmitted symbiotic bacteria in specialized cells called bacteriocytes. One of the best-studied insect nutritional endosymbioses is that of the aphid and its endosymbiont, Buchnera aphidicola. Aphids and Buchnera are metabolically and developmentally integrated, but the molecular mechanisms underlying Buchnera transmission and coordination with aphid development remain largely unknown. Previous work using electron microscopy to study aphid asexual embryogenesis has revealed that Buchnera transmission involves exocytosis from a maternal bacteriocyte followed by endocytotic uptake by a blastula...
2016: EvoDevo
https://www.readbyqxmd.com/read/27871925/identification-and-characterization-of-a-phospholipid-scramblase-encoded-by-planarian-dugesia-japonica
#11
Yu Han, Ao Li, Lili Gao, Weiwei Wu, Hongkuan Deng, Wenjing Hu, Na Li, Shimin Sun, Xiufang Zhang, Bosheng Zhao, Baohua Liu, Qiuxiang Pang
Phospholipid scramblases (PLSCRs) are the conserved calcium-binding, type II transmembrane proteins synthesized in all eukaryotic organisms. In mammals, these proteins play essential roles in various physiological processes, especially in the immune responses. However, the existence of PLSCRs and their biological functions in planarian are still unknown at present. In this study, a new member of PLSCRs was identified in planarian Dugesia japonica (D. japonica), named DjPLSCR. The sequence analysis revealed that it contains an opening reading frame consisting of 726bp encoding a putative protein of 241 amino acids with a predicted molecular mass of ~28...
February 20, 2017: Gene
https://www.readbyqxmd.com/read/27802147/inhibition-of-calcium-uptake-during-hypoxia-in-developing-zebrafish-is-mediated-by-hypoxia-inducible-factor
#12
Raymond W M Kwong, Yusuke Kumai, Velislava Tzaneva, Estelle Azzi, Nina Hochhold, Cayleih Robertson, Bernd Pelster, Steve F Perry
The present study investigated the potential role of hypoxia-inducible factor (HIF) in calcium homeostasis in developing zebrafish (Danio rerio). It was demonstrated that zebrafish raised in hypoxic water (30 mmHg; control, 155 mmHg PO2 ) until 4 days post-fertilization exhibited a substantial reduction in whole-body Ca(2+) levels and Ca(2+) uptake. Ca(2+) uptake in hypoxia-treated fish did not return to pre-hypoxia (control) levels within 2 h of transfer back to normoxic water. Results from real-time PCR showed that hypoxia decreased the whole-body mRNA expression levels of the epithelial Ca(2+) channel (ecac), but not plasma membrane Ca(2+)-ATPase (pmca2) or Na(+)/Ca(2+)-exchanger (ncx1b)...
December 15, 2016: Journal of Experimental Biology
https://www.readbyqxmd.com/read/27754889/the-zebrafish-homologs-of-set-i2pp2a-oncoprotein-expression-patterns-and-insights-into-their-physiological-roles-during-development
#13
Iliana Serifi, Eleni Tzima, Katerina Soupsana, Zoe Karetsou, Dimitris Beis, Thomais Papamarcaki
The oncoprotein SET/I2PP2A (protein phosphatase 2A inhibitor 2) 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 amino acids) and Setb (275 amino acids) proteins which share 94% identity. We show here that seta and setb are similarly expressed in the eye, the otic vesicle, the brain and the lateral line system, as indicated by in situ hybridization labeling...
December 15, 2016: Biochemical Journal
https://www.readbyqxmd.com/read/27734361/gene-expression-analysis-of-ccn-proteins-whole-mount-in-situ-hybridization-of-ccn2-in-developing-calcified-tissues
#14
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
https://www.readbyqxmd.com/read/27734351/identification-of-germ-line-stem-cells-in-zebrafish
#15
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
https://www.readbyqxmd.com/read/27734345/methods-for-studying-the-germline-of-the-human-parasite-schistosoma-mansoni
#16
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
https://www.readbyqxmd.com/read/27703040/the-salivary-protein-repertoire-of-the-polyphagous-spider-mite-tetranychus-urticae-a-quest-for-effectors
#17
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...
December 2016: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/27702788/mapping-a-multiplexed-zoo-of-mrna-expression
#18
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
https://www.readbyqxmd.com/read/27677482/characterization-of-pax3a-and-pax3b-genes-in-artificially-induced-polyploid-and-gynogenetic-olive-flounder-paralichthys-olivaceus-during-embryogenesis
#19
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
https://www.readbyqxmd.com/read/27662601/unexpected-neuroprotective-effects-of-loganin-on-1-methyl-4-phenyl-1-2-3-6-tetrahydropyridine-induced-neurotoxicity-and-cell-death-in-zebrafish
#20
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
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