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Characterization of prostaglandin F 2α receptor of mouse 3T3 fibroblasts and its functional expression in Xenopus laevis oocytes
Author(s) -
Nakao Akihide,
Watanabe Tsuyoshi,
Taniguchi Shigeo,
Nakamura Motonao,
Honda ZenIchiro,
Shimizu Takao,
Kurokawa Kiyoshi
Publication year - 1993
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.1041550206
Subject(s) - xenopus , extracellular , biology , phospholipase c , receptor , microbiology and biotechnology , alpha (finance) , inositol , endocrinology , medicine , biochemistry , gene , construct validity , nursing , patient satisfaction
Prostaglandin (PG) F 2α increased [ 3 H]thymidine incorporation into quiescent NIH 3T3 cells, stimulated phosphoinositide breakdown, and raised intracellular Ca 2+ concentration ([Ca 2+ ] i ) in a dose‐dependent manner with ED 50 values of 2.0 × 10 −8 M, 4.6 × 10 −8 M, and 7.5 × 10 −8 M, respectively. The increase in [ 3 H]thymidine incorporation with PGF 2α was additive with that seen with epidermal growth factor (EGF) or insulin. The peak [Ca 2+ ] i increase with PGF 2α was still obvious in the absence of extracellular Ca 2+ and was insensitive to islet activating protein (IAP) pretreatment. Membranes prepared from NIH 3T3 cells exhibited a specific binding for PGF 2α , which was sensitive to GTP γ S but not sensitive to IAP pretreatment. Xenopus laevis oocytes injected with NIH 3T3 cell mRNA between 18S and 28S rRNA fractionated by sucrose gradient, expressed a PGF 2α ‐specific Cl − current when examined by voltage clamp. This Cl − current was also insensitive to IAP pretreatment and not affected by extracellular Ca 2+ concentration ([Ca 2+ ] o ). These results indicate 1) that the NIH 3T3 cells expressed a specific PGF 2α receptor which is linked to phosphoinositide‐specific phospholipase C (PLC) activation and to mobilization of Ca 2+ via an IAP‐insensitive G‐proteins(s), 2) that this PGF 2α receptor may play an active role in the proliferation of NIH 3T3 cells, and 3) that this PGF 2α receptor can be expressed in the oocyte system. © 1993 Wiley‐Liss, Inc.