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Abstract
Wnt/Wg genes play a critical role in the development of various organisms. For example, the Wnt/beta-catenin signal promotes heart formation and cardiomyocyte differentiation in mice. Previous studies have shown that RGS19 (regulator of G protein signaling 19), which has Galpha subunits with GTPase activity, inhibits the Wnt/beta-catenin signal through inactivation of Galpha(o). In the present study, the effects of RGS19 on mouse cardiac development were observed. In P19 teratocarcinoma cells with RGS19 overexpression, RGS19 inhibited cardiomyocyte differentiation by blocking the Wnt signal. Additionally, several genes targeted by Wnt were down-regulated. For the in vivo study, we generated RGS19-overexpressing transgenic (RGS19 TG) mice. In these transgenic mice, septal defects and thin-walled ventricles were observed during the embryonic phase of development, and the expression of cardiogenesis-related genes, BMP4 and Mef2C, was reduced significantly. RGS19 TG mice showed increased expression levels of brain natriuretic peptide and beta-MHC, which are markers of heart failure, increase of cell proliferation, and electrocardiogram analysis shows abnormal ventricle repolarization. These data provide in vitro and in vivo evidence that RGS19 influenced cardiac development and had negative effects on heart function.
Highlights
The mammalian regulator of G protein signaling (RGS)3 family consists of Ͼ20 members [1]
This study demonstrates that the overexpression of RGS19 by P19 cells inhibited cardiomyocyte differentiation in vitro
The genes regulated by Wnt3a in an RGS19-overexpressing cell line decreased, compared with control cells
Summary
5Ј-tctgacagccagtatgacac-3Ј; reverse, 5Ј-atataggaccctacctagca-3Ј Forward, 5Ј-ggcaaggcttcactgaagac-3Ј; reverse, 5Ј-aggcaacatctctgaggcat-3Ј Forward, 5Ј-tcctgaagaacaactgggta-3Ј; reverse, 5Ј-aagtagacaccctgctcaga-3Ј Forward, 5Ј-acctcagtcagatgaaggct-3Ј; reverse, 5Ј-agtcccattacctgaggtga-3Ј Forward, 5Ј-aaccgaatgctgatggtcgt-3Ј; reverse, 5Ј-acctcattctctgggatgct-3Ј Forward, 5Ј-agcatcacggtggaggatat-3Ј; reverse, 5Ј-agaagaggatgctctctgct-3Ј Forward, 5Ј-tccaactgctctgatggcat-3Ј; reverse, 5Ј-tgctgctgcagtcgatctct-3Ј Forward, 5Ј-tgctcgaggagatgcgagat-3Ј; reverse, 5Ј-tcttgagctctgagcactca-3Ј Forward, 5Ј-tacagactctgatcgaggct-3Ј; reverse, 5Ј-ttggtcttcattcaggtggt-3Ј Forward, 5Ј-acagcaatcctgtcagcaca-3Ј; reverse, 5Ј-actgatggcatcgtgttctt-3Ј Forward, 5Ј-tggacttctggtttgcttgt-3Ј; reverse, 5Ј-acgtccactcctcttcttca-3Ј Forward, 5Ј-tggcttgcaggatacagaaa-3Ј; reverse, 5Ј-acaggtaagtgccacactga-3Ј Forward, 5Ј-agctgtttgaaggctggatt-3Ј; reverse, 5Ј-tctcctccaagtaactcggt-3Ј Forward, 5Ј-aacctgctagaccacctgga-3Ј; reverse, 5Ј-aagctgcgtgacacaccaca-3Ј Forward, 5Ј-aagacaccagtgcacaagct-3Ј; reverse, 5Ј-agccaggaggtcttcctaca-3Ј Forward, 5Ј-tgaccagctgatcaagaaca-3Ј; reverse, 5Ј-agcttcttctgcagctgact-3Ј. Amplicon size 378 bp 531 bp 329 bp 591 bp 446 bp 285 bp 370 bp 540 bp 371 bp 340 bp 385 bp 302 bp 387 bp 420 bp 462 bp 599 bp and Xenopus, whereas it enhances cardiogenesis in Drosophila and a mouse teratocarcinoma cell line. RGS19 inhibits G␣o subunit activation by the Wnt signal. Expression of RGS19 attenuates Dvl phosphorylation, -catenin accumulation, and Wnt-responsive gene transcription and blocks Wnt-induced differentiation of mouse F9 teratocarcinoma cells by inactivation of G␣o. We examined the potential mechanisms involved in attenuating Wnt signaling during cardiac muscle differentiation. Transgenic mice that overexpressed RGS19 were produced to elucidate the role of RGS19 mouse cardiac development. Cardiomyocyte differentiation in vitro and analysis of cardiac development and ventricle repolarization in mice were performed to examine the roles of RGS19
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