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Mol. Cells 2006; 21(1): 42-51

Published online January 1, 1970

© The Korean Society for Molecular and Cellular Biology

Sphingosine 1-Phosphate-induced Signal Transduction in Cat Esophagus Smooth Muscle Cells

Hyun Ju Song, Tai Sik Choi, Fa Yong Chung, Sun Young Park, Jung Soo Ryu, Jae Gwang Woo, Young Sil Min, Chang Yell Shin, Uy Dong Sohn

Abstract

We investigated the mechanism of contraction induced by S1P in esophageal smooth muscle cells. Western blot analysis demonstrated that S1P1, S1P2, S1P3, and S1P5 receptors existed in the cat esophagus. Only penetration of EDG-5 (S1P2) antibody into permeabilized cells inhibited S1P-induced contraction. Pertussis toxin (PTX) also inhibited contraction, suggesting that it was mediated by S1P2 receptors coupled to a PTX-sensitive Gi protein. Specific antibodies to Gi2, Gq and Gb inhibited contraction, implying that the S1P-induced contraction depends on PTX-insensitive Gq and Gb dimers as well as the PTX-sensitive Gi2. Contraction was not affected by the phospholipase A2 inhibitor DEDA, or the PLD inhibitor ρ-chloromer-curibenzoate, but it was abolished by the PLC inhibitor U73122. Incubation of permeabilized cells with PLCb3 antibody also inhibited contraction. Contraction involved the activation of a PKC pathway since it was affected by GF109203X and chelerythrine. Since PKCe antibody inhibited contraction, PKCe may be required. Preincubation of the muscle cells with the MEK inhibitor PD98059 blocked S1P-induced contraction, but the p38 MAP kinase inhibitor SB202190 did not. In addition, co-treatment of cells with GF 109203X and PD98059 did not have a synergistic effect, suggesting that these two kinases are involved in the same signaling pathway. Our data suggest that S1P-induced contraction in esophageal smooth muscle cells is mediated by S1P2 receptors coupled to PTX-sensitive Gi2 proteins, and PTX-insensitive Gq and Gb proteins, and that the resulting activation of the PLCb3 and PKCe pathway leads to activation of a p44/p42 MAPK pathway.

Keywords esophageal smooth muscle cell, ERK, S1P, contraction

Article

Research Article

Mol. Cells 2006; 21(1): 42-51

Published online February 28, 2006

Copyright © The Korean Society for Molecular and Cellular Biology.

Sphingosine 1-Phosphate-induced Signal Transduction in Cat Esophagus Smooth Muscle Cells

Hyun Ju Song, Tai Sik Choi, Fa Yong Chung, Sun Young Park, Jung Soo Ryu, Jae Gwang Woo, Young Sil Min, Chang Yell Shin, Uy Dong Sohn

Abstract

We investigated the mechanism of contraction induced by S1P in esophageal smooth muscle cells. Western blot analysis demonstrated that S1P1, S1P2, S1P3, and S1P5 receptors existed in the cat esophagus. Only penetration of EDG-5 (S1P2) antibody into permeabilized cells inhibited S1P-induced contraction. Pertussis toxin (PTX) also inhibited contraction, suggesting that it was mediated by S1P2 receptors coupled to a PTX-sensitive Gi protein. Specific antibodies to Gi2, Gq and Gb inhibited contraction, implying that the S1P-induced contraction depends on PTX-insensitive Gq and Gb dimers as well as the PTX-sensitive Gi2. Contraction was not affected by the phospholipase A2 inhibitor DEDA, or the PLD inhibitor ρ-chloromer-curibenzoate, but it was abolished by the PLC inhibitor U73122. Incubation of permeabilized cells with PLCb3 antibody also inhibited contraction. Contraction involved the activation of a PKC pathway since it was affected by GF109203X and chelerythrine. Since PKCe antibody inhibited contraction, PKCe may be required. Preincubation of the muscle cells with the MEK inhibitor PD98059 blocked S1P-induced contraction, but the p38 MAP kinase inhibitor SB202190 did not. In addition, co-treatment of cells with GF 109203X and PD98059 did not have a synergistic effect, suggesting that these two kinases are involved in the same signaling pathway. Our data suggest that S1P-induced contraction in esophageal smooth muscle cells is mediated by S1P2 receptors coupled to PTX-sensitive Gi2 proteins, and PTX-insensitive Gq and Gb proteins, and that the resulting activation of the PLCb3 and PKCe pathway leads to activation of a p44/p42 MAPK pathway.

Keywords: esophageal smooth muscle cell, ERK, S1P, contraction

Mol. Cells
Feb 28, 2023 Vol.46 No.2, pp. 69~129
COVER PICTURE
The bulk tissue is a heterogeneous mixture of various cell types, which is depicted as a skein of intertwined threads with diverse colors each of which represents a unique cell type. Single-cell omics analysis untangles efficiently the skein according to the color by providing information of molecules at individual cells and interpretation of such information based on different cell types. The molecules that can be profiled at the individual cell by single-cell omics analysis includes DNA (bottom middle), RNA (bottom right), and protein (bottom left). This special issue reviews single-cell technologies and computational methods that have been developed for the single-cell omics analysis and how they have been applied to improve our understanding of the underlying mechanisms of biological and pathological phenomena at the single-cell level.

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