scholarly journals High-level stable expression of recombinant 5-HT1A 5-hydroxytryptamine receptors in Chinese hamster ovary cells

1992 ◽  
Vol 285 (3) ◽  
pp. 933-938 ◽  
Author(s):  
A Newman-Tancredi ◽  
R Wootton ◽  
P G Strange
Keyword(s):  
Adenylate Cyclase ◽  
Cell Line ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Receptor Gene ◽  
Chinese Hamster ◽  
Receptor Expression ◽  
Ovary Cells ◽  
Recombinant Cell ◽  
High Level

The human 5-hydroxytryptamine 5-HT1A receptor gene was transfected into Chinese hamster ovary cells. A series of recombinant monoclonal cell lines expressing the receptor were isolated and the properties of one cell line that expressed receptors at a high level (2.8 pmol/mg) were studied in detail. In ligand binding assays with the selective 5-HT1A receptor agonist 2-(NN-di[3H]propylamino)-8-hydroxy-1,2,3,4-tetrahydronaphthalene ([3H]8-OH-DPAT) only a single class of saturable high-affinity binding sites was detected, with a pharmacological profile in competition experiments essentially identical to that of the 5-HT1A receptor of bovine hippocampus. [3H]8-OH-DPAT binding to the recombinant cell membranes was inhibited by GTP, showing that the receptors in the transfected cells couple to G-proteins. A series of 5-hydroxytryptamine agonists inhibited forskolin-stimulated adenylate cyclase activity in the cells and, despite the high level of receptor expression, their apparent efficacies were similar to those observed for inhibition of adenylate cyclase in brain. This recombinant cell line provides a complete model system for studying the 5-HT1A receptor and its transmembrane signalling system. The recombinant cells can also be grown in suspension culture for long periods but, whereas 5-HT1A receptor numbers and receptor regulation by guanine nucleotides are maintained in suspension-grown cells, the inhibition of adenylate cyclase by the 5-HT1A receptor is gradually lost.

High-Level Expression of TGF-β2 and the TGF-β2(414) Precursor in Chinese Hamster Ovary Cells

1990 ◽  
Vol 3 (2) ◽  
pp. 129-138 ◽  
Author(s):  
Linda Madisen ◽  
Mario N. Lioubin ◽  
Hans Marquardt ◽  
A. F. Purchio
Keyword(s):  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
High Level Expression ◽  
Ovary Cells ◽  
High Level

scholarly journals Identification of a Potential Receptor for Both Peptide Histidine Isoleucine and Peptide Histidine Valine

Endocrinology ◽  
2002 ◽  
Vol 143 (4) ◽  
pp. 1327-1336 ◽  
Author(s):  
Dicky Lai-Yin Tse ◽  
Ronald Ting-Kai Pang ◽  
Anderson On-Lam Wong ◽  
Siu-Ming Chan ◽  
Hubert Vaudry ◽  
...  
Keyword(s):  
Gall Bladder ◽  
Adenylate Cyclase ◽  
Chinese Hamster Ovary Cells ◽  
Receptor Gene ◽  
Chinese Hamster ◽  
Functional Expression ◽  
Coding Region ◽  
Ovary Cells ◽  
Pituitary Adenylate Cyclase ◽  
First Time

Abstract Peptide histidine isoleucine (PHI), peptide histidine valine (PHV), and vasoactive intestinal polypeptide (VIP) are cosynthesized from the same precursor and share high levels of structural similarities with overlapping biological functions. In this study, the first PHI/PHV receptor was isolated and characterized in goldfish. To study this receptor using homologous peptides, we have also characterized the goldfish prepro-PHI/VIP, and, surprisingly, a shorter transcript lacking the VIP coding region was isolated. A PHI/VIP precursor without the VIP coding sequence has never before been reported. Initial functional expression of the PHI/PHV receptor in Chinese hamster ovary cells revealed that it could be activated by human PHV [50% effective concentration (EC50): 43 nm] and to a lesser extent human PHI (EC50: 133 nm) and helodermin (EC50: 166 nm) but not fish and mammalian pituitary adenylate cyclase-activating polypeptides and VIPs. Subsequent studies indicated that, similar to the pituitary adenylate cyclase-activating polypeptide receptors (PAC1-R, VPAC1-R, and VPAC2-R), the receptor isolated in this study is able to interact with goldfish PHI and its C-terminally extended form, PHV with EC50 values 93 and 43 nm, respectively. Northern blot and RT-PCR/Southern blot analyses revealed that the PHI/VIP gene is expressed in the intestine, brain, and gall bladder and the PHI/PHV receptor gene is primarily expressed in the pituitary and to a lesser extend in the intestine and gall bladder, suggesting that PHI/PHV may play a role, notably in the regulation of pituitary function. In conclusion, our results demonstrate for the first time the existence of a PHI/PHV receptor, indicating that the functions of PHI and PHV could be mediated by their own receptor in addition to VIP receptors.

scholarly journals Enhancement of cytotoxicities of ricin and Pseudomonas toxin in Chinese hamster ovary cells by nigericin.

1981 ◽  
Vol 1 (6) ◽  
pp. 552-559 ◽  
Author(s):  
B Ray ◽  
H C Wu
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Protein Synthesis ◽  
Cell Line ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Diphtheria Toxin ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Ovary Cells ◽  
Pseudomonas Aeruginosa Exotoxin

Nigericin and monensin, ionophores for Na+ and K+, have been found to enhance the cytotoxicities of abrin, ricin, and Pseudomonas aeruginosa exotoxin A in Chinese hamster ovary (CHO) cells. They do not affect the cytotoxicity of diphtheria toxin in the same cell line. Maximal sensitization of the CHO cells toward ricin and Pseudomonas toxin requires preculture of CHO cells in the presence of nigericin. Inhibition of protein synthesis in CHO cells by ricin or Pseudomonas toxin is also enhanced by preculture of CHO cells in the presence of nigericin. These results suggest a common step in the intoxication process of ricin and Pseudomonas toxin, the rate of which is facilitated by pretreatment with nigericin. This step is, however, not shared by the intoxication of CHO cells with diphtheria toxin.

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