scholarly journals Sequence analysis and spatiotemporal developmental distribution of the Cat-1-type transporter slc7a1a in zebrafish (Danio rerio)

2020 ◽  
Vol 46 (6) ◽  
pp. 2281-2298
Author(s):  
Ståle Ellingsen ◽  
Shailesh Narawane ◽  
Anders Fjose ◽  
Tiziano Verri ◽  
Ivar Rønnestad
Keyword(s):  
Sequence Analysis ◽  
Danio Rerio ◽  
Tissue Expression ◽  
Biological Processes ◽  
Cationic Amino Acid ◽  
Branchial Arches ◽  
Whole Mount ◽  
Cationic Amino Acids ◽  
The Central Nervous System

Abstract Cationic amino acid transporter 1 (Cat-1 alias Slc7a1) is a Na+-independent carrier system involved in transport and absorption of the cationic amino acids lysine, arginine, histidine, and ornithine and has also been shown to be indispensable in a large variety of biological processes. Starting from isolated full-length zebrafish (Danio rerio) cDNA for slc7a1a, we performed comparative and phylogenetic sequence analysis, investigated the conservation of the gene during vertebrate evolution, and defined tissue expression during zebrafish development. Whole mount in situ hybridization first detected slc7a1a transcripts in somites, eyes, and brain at 14 h post-fertilization (hpf) with additional expression in the distal nephron at 24 hpf and in branchial arches at 3 days post-fertilization (dpf), with significant increase by 5 dpf. Taken together, the expression analysis of the zebrafish Cat-1 system gene slc7a1a suggests a functional role(s) during the early development of the central nervous system, muscle, gills, and kidney.

scholarly journals A Systems Approach and Skeletal Myogenesis

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Yoshiaki Ito ◽  
Tomohiro Kayama ◽  
Hiroshi Asahara
Keyword(s):  
Gene Regulation ◽  
Regulatory Network ◽  
High Throughput Screening ◽  
Systems Approach ◽  
Biological Processes ◽  
Skeletal Myogenesis ◽  
Genome Wide ◽  
Whole Mount ◽  
Strict Regulation

Skeletal myogenesis depends on the strict regulation of the expression of various gene subsets. Therefore, the understanding of genome wide gene regulation is imperative for elucidation of skeletal myogenesis. In recent years, systems approach has contributed to the understanding of various biological processes. Our group recently revealed the critical genome network of skeletal myogenesis by using a novel systems approach combined with whole-mountin situhybridization (WISH) database, high-throughput screening, and microarray analysis. In this paper, we introduce our systems approach for understanding the myogenesis regulatory network and describe the advantages of systems approach.

Cloning, sequence analysis, and tissue expression of marmoset paraoxonase 1

2021 ◽  
pp. 100398
Author(s):  
Shotaro Uehara ◽  
Yasuhiro Uno ◽  
Makiko Shimizu ◽  
Hiroshi Yamazaki
Keyword(s):  
Sequence Analysis ◽  
Tissue Expression ◽  
Paraoxonase 1

scholarly journals Molecular cloning and biochemical characterization of sialidases from zebrafish (Danio rerio)

2007 ◽  
Vol 408 (3) ◽  
pp. 395-406 ◽  
Author(s):  
Marta Manzoni ◽  
Paolo Colombi ◽  
Nadia Papini ◽  
Luana Rubaga ◽  
Natascia Tiso ◽  
...  
Keyword(s):  
Danio Rerio ◽  
Biochemical Characterization ◽  
Chromosome 21 ◽  
Ph Optimum ◽  
Reverse Transcription Pcr ◽  
Genome Wide ◽  
Genome Wide Search ◽  
Putative Orthologues

Sialidases remove sialic acid residues from various sialo-derivatives. To gain further insights into the biological roles of sialidases in vertebrates, we exploited zebrafish (Danio rerio) as an animal model. A zebrafish transcriptome- and genome-wide search using the sequences of the human NEU polypeptides as templates revealed the presence of seven different genes related to human sialidases. neu1 and neu4 are the putative orthologues of the mammalian sialidases NEU1 and NEU4 respectively. Interestingly, the remaining genes are organized in clusters located on chromosome 21 and are all more closely related to mammalian sialidase NEU3. They were thus named neu3.1, neu3.2, neu3.3, neu3.4 and neu3.5. Using RT–PCR (reverse transcription–PCR) we detected transcripts for all genes, apart from neu3.4, and whole-mount in situ hybridization experiments show a localized expression pattern in gut and lens for neu3.1 and neu4 respectively. Transfection experiments in COS7 (monkey kidney) cells demonstrate that Neu3.1, Neu3.2, Neu3.3 and Neu4 zebrafish proteins are sialidase enzymes. Neu3.1, Neu3.3 and Neu4 are membrane-associated and show a very acidic pH optimum below 3.0, whereas Neu3.2 is a soluble sialidase with a pH optimum of 5.6. These results were further confirmed by subcellular localization studies carried out using immunofluorescence. Moreover, expression in COS7 cells of these novel zebrafish sialidases (with the exception of Neu3.2) induces a significant modification of the ganglioside pattern, consistent with the results obtained with membrane-associated mammalian sialidases. Overall, the redundancy of sialidases together with their expression profile and their activity exerted on gangliosides of living cells indicate the biological relevance of this class of enzymes in zebrafish.

scholarly journals FISHing for chick genes: Triple-label whole-mount fluorescence in situ hybridization detects simultaneous and overlapping gene expression in avian embryos

2004 ◽  
Vol 229 (3) ◽  
pp. 651-657 ◽  
Author(s):  
Nathaniel Denkers ◽  
Pilar García-Villalba ◽  
Christopher K. Rodesch ◽  
Kandice R. Nielson ◽  
Teri Jo Mauch
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Avian Embryos ◽  
Overlapping Gene ◽  
Whole Mount

Mouse Wnt genes exhibit discrete domains of expression in the early embryonic CNS and limb buds

Development ◽  
1993 ◽  
Vol 119 (1) ◽  
pp. 247-261 ◽  
Author(s):  
B.A. Parr ◽  
M.J. Shea ◽  
G. Vassileva ◽  
A.P. McMahon
Keyword(s):  
Limb Development ◽  
Expression Patterns ◽  
Limb Buds ◽  
Expression Studies ◽  
The Family ◽  
The Central Nervous System ◽  
Discrete Domains ◽  
Early Developmental ◽  
The Brain

Mutation and expression studies have implicated the Wnt gene family in early developmental decision making in vertebrates and flies. In a detailed comparative analysis, we have used in situ hybridization of 8.0- to 9.5-day mouse embryos to characterize expression of all ten published Wnt genes in the central nervous system (CNS) and limb buds. Seven of the family members show restricted expression patterns in the brain. At least three genes (Wnt-3, Wnt-3a, and Wnt-7b) exhibit sharp boundaries of expression in the forebrain that may predict subdivisions of the region later in development. In the spinal cord, Wnt-1, Wnt-3, and Wnt-3a are expressed dorsally, Wnt-5a, Wnt-7a, and Wnt-7b more ventrally, and Wnt-4 both dorsally and in the floor plate. In the forelimb primordia, Wnt-3, Wnt-4, Wnt-6 and Wnt-7b are expressed fairly uniformly throughout the limb ectoderm. Wnt-5a RNA is distributed in a proximal to distal gradient through the limb mesenchyme and ectoderm. Along the limb's dorsal-ventral axis, Wnt-5a is expressed in the ventral ectoderm and Wnt-7a in the dorsal ectoderm. We discuss the significance of these patterns of restricted and partially overlapping domains of expression with respect to the putative function of Wnt signalling in early CNS and limb development.

Mouse Hox-3.4: homeobox sequence and embryonic expression patterns compared with other members of the Hox gene network

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 329-339 ◽  
Author(s):  
S.J. Gaunt ◽  
P.L. Coletta ◽  
D. Pravtcheva ◽  
P.T. Sharpe
Keyword(s):  
Gene Network ◽  
Common Ancestor ◽  
Expression Patterns ◽  
Homeobox Gene ◽  
Predicted Amino Acid Sequence ◽  
Chromosome 15 ◽  
Hox Gene ◽  
The Central Nervous System ◽  
Genomic Organisation

A putative mouse homeobox gene (Hox-3.4) was previously identified 4kb downstream of the Hox-3.3 (Hox-6.1)* gene (Sharpe et al. 1988). We have now sequenced the Hox-3.4 homeobox region. The predicted amino acid sequence shows highest degree of homology in the mouse with Hox-1.3 and -2.1. This, together with similarities in the genomic organisation around these three genes, suggests that they are comembers of a subfamily, derived from a common ancestor. Hox-3.4 appears to be a homologue of the Xenopus Xlhbox5 and human cp11 genes (Fritz and De Robertis, 1988; Simeone et al. 1988). Using a panel of mouse-hamster somatic cell hybrids we have mapped the Hox-3.4 gene to chromosome 15. From the results of in situ hybridization experiments, we describe the distribution of Hox-3.4 transcripts within the 12 1/2 day mouse embryo, and we compare this with the distributions of transcripts shown by seven other members of the Hox gene network. We note three consistencies that underlie the patterns of expression shown by Hox-3.4. First, the anterior limits of Hox-3.4 transcripts in the embryo are related to the position of the Hox-3.4 gene within the Hox-3 locus. Second, the anterior limits of Hox-3.4 expression within the central nervous system are similar to those shown by subfamily homologues Hox-2.1 and Hox-1.3, although the tissue-specific patterns of expression for these three genes show many differences. Third, the patterns of Hox-3.4 expression within the spinal cord and the testis are very similar to those shown by a neighbouring Hox-3 gene (Hox-3.3), but they are quite different from those shown by Hox-1 genes (Hox-1.2, -1.3 and -1.4).

scholarly journals Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells.

1990 ◽  
Vol 110 (6) ◽  
pp. 2117-2132 ◽  
Author(s):  
A Taraboulos ◽  
D Serban ◽  
S B Prusiner
Keyword(s):  
Cultured Cells ◽  
Prion Diseases ◽  
Neuroblastoma Cells ◽  
Brain Cell ◽  
Cellular Prion Protein ◽  
Intracellular Accumulation ◽  
Golgi Stack ◽  
Glycosyl Phosphatidylinositol ◽  
The Central Nervous System

The cellular prion protein (PrPC) is a sialoglycoprotein anchored to the external surface of cells by a glycosyl phosphatidylinositol moiety. During scrapie, an abnormal PrP isoform designated PrPSc accumulates, and much evidence argues that it is a major and necessary component of the infectious prion. Based on the resistance of native PrPSc to proteolysis and to digestion with phosphatidylinositol-specific phospholipase C as well as the enhancement of PrPSc immunoreactivity after denaturation, we devised in situ immunoassays for the detection of PrPSc in cultured cells. Using these immunoassays, we identified the sites of PrPSc accumulation in scrapie-infected cultured cells. We also used these immunoassays to isolate PrPSc-producing clones from a new hamster brain cell line (HaB) and found an excellent correlation between their PrPSc content and prion infectivity titers. In scrapie-infected HaB cells as well as in scrapie-infected mouse neuroblastoma cells, most PrPSc was found to be intracellular and most localized with ligands of the Golgi marker wheat germ agglutinin. In one scrapie-infected HaB clone, PrPSc also localized extensively with MG-160, a protein resident of the medial-Golgi stack whereas this colocalization was not observed in another subclone of these cells. Whether the sites of intracellular accumulation of PrPSc are limited to a few subcellular organelles or they are highly variable remains to be determined. If the intracellular accumulation of PrPSc is found in the cells of the central nervous system, then it might be responsible for the neuronal dysfunction and degeneration which are cardinal features of prion diseases.

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