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Identification of an IP 3 receptor in endothelial cells
Author(s) -
Bourguig Lilly Y. W.,
Iida N.,
Sobrin L.,
Bourguig C. J.
Publication year - 1994
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041590105
Subject(s) - receptor , endothelial stem cell , polyclonal antibodies , biology , microbiology and biotechnology , agonist , cell culture , antibody , chemistry , in vitro , biochemistry , immunology , genetics
In this study we have used saponin to permeabilize bovine endothelial cell membranes in order to directly test the involvement of IP 3 in regulating internal Ca 2+ release. Our results indicate that the release of internal Ca 2+ occurs as early as 1–3 seconds after IP 3 addition. This IP 3 ‐induced internal Ca 2+ release can be inhibited by heparin (an IP 3 receptor antagonist). Further binding of [ 3 H]IP 3 to saponin‐permeabilized bovine endothelial cells reveals the presence of a single, high affinity class of IP 3 receptor with a dissociation constant (K d ) of ≈0.50 (±0.03) nM. Using a panel of monoclonal and polyclonal antibodies against IP 3 receptor, we have established that the bovine endothelial cell IP 3 receptor (≈260 kDa) displays immunological cross‐reactivity with the rat brain IP 3 receptor. Immunofluorescence data indicates that the IP 3 receptor is preferentially located at the perinuclear region of the cells. In addition, PCR analysis of first‐strand cDNAs from both bovine endothelial cells and rat brain tissues reveals that the IP 3 receptor transcript in bovine endothelial cells belongs to the short non‐neuronal form and not the long neuronal form detected in rat brain tissue. These findings suggest that the IP 3 receptor in endothelial cells is both structurally and functionally analogous to that reported in non‐neuronal cell systems and probably plays an important role in agonist‐induced endothelial cell activation. © 1994 wiley‐Liss, Inc.