Characterization of vasopressin-induced calcium mobilization in CHO cells transfected rat V1a receptor

Small polydisperse circular (spc) DNA was isolated and cloned, using BglII from Chinese hamster ovary (CHO) cells. The properties of 47 clones containing at least 43 different BglII fragments are reported. The majority of the clones probably contain entire sequences from individual spcDNA molecules. Most of the clones were homologous to sequences in CHO cell chromosomal DNA, and many were also homologous to mouse LMTK- cell chromosomal sequences. The majority of homologous CHO cell chromosomal sequences were repetitive, although a few may be single copy. Only a small fraction of cloned spcDNA molecules were present in every cell; most occurred less frequently than once in 15 cells. Localization studies indicated that at least a portion of spcDNA is associated with the nucleus in CHO cells.

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Identification and functional characterization of a novel human protein highly related to the yeast dynamin-like GTPase Vps1p

Journal of Cell Science ◽

10.1242/jcs.111.10.1341 ◽

1998 ◽

Vol 111 (10) ◽

pp. 1341-1349 ◽

Cited By ~ 3

Author(s):

M. Imoto ◽

I. Tachibana ◽

R. Urrutia

Keyword(s):

Endoplasmic Reticulum ◽

Cho Cells ◽

Mammalian Cells ◽

Functional Characterization ◽

Luciferase Reporter ◽

Pleckstrin Homology ◽

Rich Region ◽

Gtpase Domain ◽

Proline Rich Region

Dynamin proteins containing a GTPase domain, a pleckstrin homology motif and a proline-rich tail participate in receptor-mediated endocytosis in organisms ranging from insects to vertebrates. In addition, dynamin-related GTPases, such as the yeast Golgi protein Vps1p, which lack both the pleckstrin homology motif and the proline-rich region, participate in vesicular transport within the secretory pathway in lower eukaryotes. However, no data is available on the existence of Vps1p-like proteins in mammalian cells. In this study, we report the identification and characterization of a novel gene encoding a human dynamin-related protein, DRP1, displaying high similarity to the Golgi dynamin-like protein Vps1p from yeast and to a Caenorhabditis elegans protein deposited in the databank. These proteins are highly conserved in their N-terminal tripartite GTPase domain but lack the pleckstrin homology motif and proline-rich region. Northern blot analysis reveals that the DRP1 mRNA is detected at high levels in human muscle, heart, kidney and brain. Immunolocalization studies in Chinese hamster ovary (CHO) cells using an epitope-tagged form of DRP1 and confocal microscopy show that this protein is concentrated in a perinuclear region that labels with the endoplasmic reticulum marker DiOC6(3) and the Golgi marker C5-DMB-Cer. In addition, the localization of DRP1 is highly similar to the localization of the endoplasmic reticulum and cis-Golgi GTPase Rab1A, but not to the staining for the trans-Golgi GTPase Rab6. Furthermore, overexpression of a cDNA encoding a GTP binding site mutant of DRP1 (DRP1(K38E)) in CHO cells decreases the amount of a secreted luciferase reporter protein, whereas the overexpression of wild-type DRP1 increases the secretion of this marker. Together, these results constitute the first structural and functional characterization of a mammalian protein similar to the yeast dynamin-related GTPase Vps1p and indicate that the participation of these proteins in secretion has been conserved throughout evolution.

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PRODUCTION AND CHARACTERIZATION OF A RECOMBINANT FACTOR VIII EXPRESSED BY CHO CELLS CULTURED IN SERUM SUPPLEMENTED OR IN SERUM FREE MEDIA

Animal Cell Technology ◽

10.1016/b978-0-7506-0421-5.50044-1 ◽

1992 ◽

pp. 152-154

Author(s):

G. MIGNOT ◽

N. BIHOREAU ◽

P. PAOLANTONACCI ◽

A. SAUGER ◽

V. TOULLY ◽

...

Keyword(s):

Factor Viii ◽

Cho Cells ◽

Recombinant Factor Viii ◽

Serum Free ◽

Free Media ◽

Cells Cultured

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Pharmacological characterization of recombinant N-type calcium channel (Cav2.2) mediated calcium mobilization using FLIPR

Biochemical Pharmacology ◽

10.1016/j.bcp.2006.06.003 ◽

2006 ◽

Vol 72 (6) ◽

pp. 770-782 ◽

Cited By ~ 18

Author(s):

Elfrida R. Benjamin ◽

Farhana Pruthi ◽

Shakira Olanrewaju ◽

Shen Shan ◽

Denise Hanway ◽

...

Keyword(s):

Calcium Channel ◽

Calcium Mobilization ◽

Pharmacological Characterization

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Pharmacological characterization of human NPFF1 and NPFF2 receptors expressed in CHO cells by using NPY Y1 receptor antagonists

European Journal of Pharmacology ◽

10.1016/s0014-2999(02)02224-0 ◽

2002 ◽

Vol 451 (3) ◽

pp. 245-256 ◽

Cited By ~ 102

Author(s):

Catherine Mollereau ◽

Honoré Mazarguil ◽

Delphine Marcus ◽

Isabelle Quelven ◽

Masato Kotani ◽

...

Keyword(s):

Cho Cells ◽

Receptor Antagonists ◽

Pharmacological Characterization ◽

Y1 Receptor

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Isolation and characterization of Chinese hamster ovary cell variants defective in adhesion to fibronectin-coated collagen.

The Journal of Cell Biology ◽

10.1083/jcb.87.3.755 ◽

1980 ◽

Vol 87 (3) ◽

pp. 755-763 ◽

Cited By ~ 38

Author(s):

P A Harper ◽

R L Juliano

Keyword(s):

Chinese Hamster Ovary ◽

Cho Cells ◽

Divalent Cations ◽

Chinese Hamster ◽

Ovary Cell ◽

Human Diploid Fibroblasts ◽

Isolation And Characterization ◽

A Cell ◽

Cold Insoluble Globulin

Variant clones of Chinese hamster ovary (CHO) cells were selected for reduced adhesion to serum-coated tissue culture plates. These clones also displayed reduced adhesion to substrata composed of collagen layers coated with bovine serum or with fibronectin (cold-insoluble globulin). Wild-type (WT) and adhesion variant (ADv) cells grew at comparable rates in suspension culture, but the adhesion variants could not be grown in monolayer culture because of their inability to attach to the substratum. The adhesion deficit in these cells was not corrected by raising the concentration of divalent cations or of serum to levels 10-fold greater than those normally utilized in cell culture. However, both WT and ADv clones could adhere, spread, and attain a normal CHO morphology on substrata coated with concanavalin A or poly-L-lysine. In addition, the adhesion variants could attach to substrata coated with "footpad" material (substratum-attached material) derived from monolayers of human diploid fibroblasts or WT CHO cells. These observations suggest that the variant clones may have a cell surface defect that prevents them from utilizing exogeneous fibronectin as an adhesion-promoting ligand; however the variants seem to have normal cytoskeletal and metabolic capacities that allow them to attach and spread on substrata coated with alternative ligands. These variants should be extremely useful in studying the molecular basis of cell adhesion.

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Development of a Microfluidic GHz Impedance Cytometer

tm - Technisches Messen ◽

10.1515/teme.2013.0053 ◽

2013 ◽

Vol 80 (12) ◽

Cited By ~ 1

Author(s):

Niels Haandbæk ◽

Sebastian C. Bürgel ◽

Flavio Heer ◽

Andreas Hierlemann

Keyword(s):

Dielectric Properties ◽

Chinese Hamster Ovary ◽

Cho Cells ◽

Single Cells ◽

Chinese Hamster ◽

Frequency Range ◽

Cell Nuclei ◽

Dielectric Characterization ◽

The Difference

AbstractThis article presents a novel microfluidic impedance cytometer enabling dielectric characterization of single cells at frequencies up to 500 MHz. The dielectric properties of cells at lower frequencies contain information about their size and membrane capacitance. The increased frequency range of the presented cytometer potentially allows for characterization of intracellular components, such as vacuoles or the cell nuclei. We demonstrate the overall capabilities of the cytometer through discrimination of polystyrene beads from Chinese hamster ovary (CHO) cells. The discrimination is based on the difference in dielectric properties at frequencies up to 500 MHz.

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