Microarray analysis of PDGFRα+ populations in ES cell differentiation culture identifies genes involved in differentiation of mesoderm and mesenchyme including ARID3b that is essential for development of embryonic mesenchymal cells

Abstract

An inherent difficulty in using DNA microarray technology on the early mouse embryo is its relatively small size. In this study, we investigated whether use of ES cell differentiation culture, which has no theoretical limit in the number of cells that can be generated, can improve this situation. Seven distinct ES-cell-derived populations were analyzed by DNA microarray and examined for genes whose distribution patterns are similar to those of PDGFRalpha, a gene implicated in differentiation of mesoderm/mesenchymal lineages. Using software developed in our laboratory, we formed a group of 30 genes which showed the highest similarity to PDGFRalpha, 18 of these genes were shown to be involved in development of either mesodermal, mesenchymal or neural crest cells. This list also contains several genes whose role in embryogenesis has not yet been fully identified. One such molecule is mARID3b. The mARID3b expression is found in the paraxial mesoderm and cranial mesenchyme. mARID3b-null mouse showed early embryonic lethality, and most phenotypes of this mutant appear to develop from a failure to generate a sufficient number of cranial mesenchymal cells. These results demonstrate the potential use of ES cell differentiation culture in identifying novel genes playing an indispensable role in embryogenesis.

Cross Talk between Id1 and Its Interactive Protein Dril1 Mediate Fibroblast Responses to Transforming Growth Factor-β in Pulmonary Fibrosis

“…31 The highly similar (and ubiquitously expressed) protein ARID3B (Bdp) may be the true murine counterpart of Dril1. In contrast to Bright, deletion of Bdp in the mouse is lethal, 33 as predicted for Dril1. The data presented here indicate that Dril1 can alter fibroblast responses to TGF-␤ 1 and is highly expressed in adult fibrotic lung tissue, suggesting that this molecule has a functional role in the adult lung.…”

The novel protein kinase Vlk is essential for stromal function of mesenchymal cells

“…RNA extraction using Trizol (Invitrogen), cRNA probe preparation and hybridization to Affymetrix oligonucleotide arrays MGU74v2 A, B and C were performed as recommended by the manufacturers, as described previously (Takebe et al, 2006). The expression data were maintained and analyzed using the eXintegrator system (http://www.cdb.riken.jp/scb/documentation/).…”

“…In situ hybridization was performed as described previously (Takebe et al, 2006). The mouse Vlk probe was amplified by RT-PCR using primers 5Ј-TGGAGACAGCGCTACACTTG-3Ј and 5Ј-CCCAGTTAAGACCA -GCCTGA-3Ј.…”

“…Among various settings for gene mining, in vitro differentiation of embryonic stem (ES) cells towards specific lineages has emerged as a promising experimental system that has marked advantages over study of the embryo itself, from which only small numbers of cells can be obtained (Hirashima et al, 2004;Keller et al, 1993;Nishikawa et al, 1998). We have been using ES cell differentiation in combination with highthroughput technologies to identify genes that are involved during early embryogenesis (Takebe et al, 2006).…”

Mesogenin 1 is a master regulator of paraxial presomitic mesoderm differentiation

“…1I). Pdgfrα is a cell surface marker of mesenchymal paraxial mesoderm progenitors (Sakurai et al, 2006;Takebe et al, 2006), and, together with Flk1 (Kdr -Mouse Genome Informatics), marks progenitors with broad mesodermal potential (Ema et al, 2006;Lindsley et al, 2008 these mesoderm markers (Fig. 1J).…”