Flow Cytometric Analysis of Internal Calcium Mobilization via a B2-Bradykinin Receptor on a Subclone of PC-12 Cells
| Autor: | N. A. Sharif, H. M. Cherwinski, J. T. Ransom, J. F. Dunne |
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| Rok vydání: | 1991 |
| Předmět: |
Agonist
medicine.medical_specialty medicine.drug_class Agonist-antagonist Rest Population Adrenal Gland Neoplasms Bradykinin Pheochromocytoma Biology Biochemistry Flow cytometry Cellular and Molecular Neuroscience chemistry.chemical_compound Internal medicine Tumor Cells Cultured medicine Animals Receptor education education.field_of_study medicine.diagnostic_test Receptors Bradykinin Flow Cytometry Molecular biology Receptors Neurotransmitter Endocrinology chemistry Second messenger system Calcium B2 Bradykinin Receptor |
| Zdroj: | Journal of Neurochemistry. 56:983-989 |
| ISSN: | 1471-4159 0022-3042 |
| DOI: | 10.1111/j.1471-4159.1991.tb02018.x |
| Popis: | Single cell Ca2+ mobilization was studied by nonparametric, quantitative flow cytometry using a sort-selected subclone of PC-12 cells. The response of the parent PC-12 population to bradykinin (BK) was very heterogeneous and of a relatively low magnitude. Cells that exhibited maximal Ca2+ mobilization were singly sorted by flow cytometry, cultured, and reanalyzed. In one subclone, referred to as BK1, BK or the B2-BK receptor agonists Lys-BK and Met-Lys-BK (10 pM-1 microM) induced robust Ca2+ transients in 80% of the cells. All three peptides produced the same maximal responses. The B1-BK receptor agonist Des-Arg9-BK (1 nM-1 microM) failed to elicit Ca2+ mobilization in these cells. The responses to BK (10 and 100 nM) were inhibited by preincubation with the B2-receptor antagonists D-Arg0-Hyp3-thienyl5,8-D-Phe7-BK and D-Arg0-Hyp3-D-Phe7 (0.1 nM-10 microM) in a concentration-dependent manner. Des-Arg9-Leu8-BK, a B1-receptor antagonist, failed to block the BK responses at 0.1-10 microM. The agonist/antagonist profile of the BK responses indicated that the B2-BK receptor mediated the Ca2+ response in the BK1 subclone. Thus, flow cytometric analysis of a receptor-mediated Ca2+ response can be employed to select a homogeneously responsive subclone from a heterogeneous, clonal population that can improve the resolution of receptor-mediated second messenger generation at the single cell level. |
| Databáze: | OpenAIRE |
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