A mouse embryonic stem cell line showing pluripotency of differentiation in early embryos and ubiquitous β-galactosidase expression

Suemori, Hirofumi; Kadodawa, Yuzo; Nakatsuji, Norio; Goto, Koji; Araki, Isato; Kondoh, Hisato

doi:10.1016/0922-3371(90)90120-l

Cited by 174 publications

(88 citation statements)

References 18 publications

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“…The PCR product was purified over a Wizard PCR clean up column (Promega, Madison, WI), cut with HindIII, and ligated directly into the HindIII site separating the promoter and polyadenylylation signal sequences of the Miw plasmid. Miw contains a Rous sarcoma virus long terminal repeat inserted into the chicken ␤-actin promoter (14,15). The transgene was liberated from the vector by digestion with NotI and purified from a gel using the Geneclean purification system (MP Biochemicals, Solon, OH).…”

Section: Methodsmentioning

confidence: 99%

Keratinocyte-specific Expression of Fatty Acid Transport Protein 4 Rescues the Wrinkle-free Phenotype in Slc27a4/Fatp4 Mutant Mice

Moulson

Lin

White

et al. 2007

Journal of Biological Chemistry

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FATP4 (fatty acid transport protein 4; also known as SLC27A4) is the most widely expressed member of a family of six long chain fatty acid transporters. FATP4 is highly expressed in enterocytes and has therefore been proposed to be a major importer of dietary fatty acids. Two independent mutations in Fatp4 cause mice to be born with thick, tight, shiny, "wrinklefree" skin and a defective skin barrier; they die within hours of birth from dehydration and restricted movements. In contrast, induced keratinocyte-specific deficiency of FATP4 in adult mice causes only mild skin abnormalities. Therefore, whether the loss of FATP4 from skin or a systemic gestational metabolic defect causes the severe skin defects and neonatal lethality remain important unanswered questions. To investigate the basis for the phenotype, we first generated wild-type tetraploid/mutant diploid aggregates that should lead to rescue of any abnormalities caused by loss of FATP4 from the placenta. However, the skin phenotype was not ameliorated. We then generated transgenic mice expressing exogenous FATP4 either widely or specifically in suprabasal keratinocytes, and we bred the transgenes onto the Fatp4 ؊/؊ background. Both modes of FATP4 expression led to rescue of the neonatally lethal skin defects, and the resulting mice were viable and fertile. Keratinocyte expression of an FATP4 variant with mutations in the acyl-CoA synthetase domain did not provide any degree of rescue. We conclude that expression of FATP4 with an intact acyl-CoA synthetase domain in suprabasal keratinocytes is necessary for normal skin development and that FATP4 functions in establishing the cornified envelope. Fatty acid transport proteins (FATPs)2 compose a family of transmembrane proteins that facilitate cellular long chain fatty acid uptake (1-3). The most widely expressed member of this family is FATP4 (4, 5), which is encoded by Slc27a4 (solute carrier family 27 member 4 gene). The most robust expression of FATP4 is in intestinal epithelial cells (5), but its broad expression pattern is suggestive of functions in many organs.Surprisingly, the "wrinkle-free" phenotype we discovered in mice with a spontaneous mutation in Slc27a4 (Slc27a4 wrfr ) and the identical phenotype of mice with a targeted Slc27a4 mutation (Slc27a4) indicate critical roles for FATP4 in skin and hair development (6, 7). For these mutations, which both affect exon 3, homozygotes are born with tight, thick, shiny skin and a defective skin barrier; they die shortly after birth because of restricted movements and dehydration. These defects are reminiscent of human restrictive dermopathy, but this disease has recently been linked to mutations in zinc metalloproteinase STE24 (8 -10). Interestingly, deletion of Slc27a4 exons 2 and 3 (the first two coding exons; Slc27a4 tm1Asta ) results in embryonic lethality prior to embryonic day 9.5 (11). The reason for the striking difference in phenotypes remains unknown, but it may involve a partially functional truncated protein in the first two mutants or early ...

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Section: Methodsmentioning

confidence: 99%

Keratinocyte-specific Expression of Fatty Acid Transport Protein 4 Rescues the Wrinkle-free Phenotype in Slc27a4/Fatp4 Mutant Mice

Moulson

Lin

White

et al. 2007

Journal of Biological Chemistry

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“…cDNAs encoding fulllength mouse RPTP [National Center for Biotechnology Information (NCBI) accession number U55057] or RPTP-⌬IC (corresponding to nucleotides 352-2701 of NCBI accession number U55057) were fused to a triple HA-tag and cloned into the expression vector pMES, which includes an internal ribosomal entry site (IRES)-green fluorescent protein (GFP) cassette (Swartz et al, 2001) or into the vector pMIWIII, which lacks IRES-GFP (Suemori et al, 1990). The coding sequence of chick RPTP was cloned using degenerate primers based on published sequences of the mouse, rat, and human orthologs and found to be mostly identical to NCBI accession number AY147868 (sequences are available on request).…”

Section: Methodsmentioning

confidence: 99%

A Role for Receptor Protein Tyrosine Phosphatase in Midbrain Development

Badde

Schulte

2008

Journal of Neuroscience

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The mid-hindbrain boundary (MHB) harbors an important organizing center for the adjacent brain regions. Here, we present evidence that the receptor protein tyrosine phosphatase (RPTP) is part of the complex molecular network that maintains and shapes the MHB region. RPTP is expressed in a tight band of cells in the caudal midbrain, anterior to the transverse ring of Wnt1 expression. Forced expression of RPTP across the mid-hindbrain region repressed expression of Wnt1, whereas RNA interference-mediated knock-down of RPTP resulted in expansion and distortion of the Wnt1 domain. When ectopically expressed in the mesencephalon, RPTP specifically inhibited the induction of Wnt1 expression after subsequent stimulation with Fgf8. Reduced Wnt1 expression after RPTP transfection correlated with a decrease in Ras-mitogen-activated protein kinase activity at the MHB. We further show that in the embryonic midbrain, RPTP can bind to ␤-catenin, a central component of the canonical Wnt signaling pathway. Overexpression of RPTP suppressed the activity of a ␤-catenin responsive promoter in the midbrain and reduced progenitor cell proliferation. Cotransfection of Wnt1 or of a stabilized form of ␤-catenin together with RPTP partially rescued the RPTP-mediated proliferation defect. Together, these data suggest that RPTP may play a dual role in the control of midbrain development: as a negative modulator of Fgf8-induced Wnt1 expression at the MHB, which may help to confine the Wnt1 domain to it characteristic tight ring at the MHB; and as an inhibitor of canonical Wnt signaling through interaction with and presumably sequestration of ␤-catenin.

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“…Plasmid Construction-To construct expression vectors for transfection, fragments containing full-length coding regions of chick Hoxa-13, Hoxd , and Hoxa-4 were subcloned into the pMiw expression vector (27), which contained the Rous sarcoma virus long terminal repeat enhancer and chicken ␤-actin promoter. To generate several deletion constructs of HoxA-13, pMiwHoxA-13 plasmid was digested with pairs of restriction enzymes, EcoRV and BamHI (aa 1-272; A13-dC33), AatII and BamHI (aa 1-227; A13-dHD), NcoI and EcoRV (aa 1-191; A13-N191), and NcoI and AatII (aa 1-191 and 225-305; A13-dAD) followed by internal ligation or oligolinker-mediated ligation.…”

Section: Methodsmentioning

confidence: 99%

Hox Proteins Functionally Cooperate with the GC Box-binding Protein System through Distinct Domains

Suzuki

Ueno

Kuroiwa

2003

Journal of Biological Chemistry

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Hox genes encode a transcriptional factor that plays a key role in regulating position-specific cartilage pattern formation. We found that Hoxa-13 and Hoxd-13, which are members of the Abd-B subfamily of Hox genes and are crucial for the autopod development of the limb, stimulate transcription from the Bmp-4 promoter. This stimulation was dependent on the GC box within the promoter and independent of the putative Hox protein binding site. The stimulation by HoxA-13 was remarkably enhanced by cotransfection with members of a family of zinc finger GC box binding transcriptional factors including Sp1. The stimulation was suppressed by another Abd-B Hox protein, HoxA-11, indicating that each Abd-B Hox protein has a different effect on the target genes through the Sp1 system. We have identified multiple functional domains involved in transcriptional regulation, including three independent transcriptional activation domains (ADs) in HoxA-13. AD1 and AD3 in helices 1 and 2 of the homeodomain individually cooperate with Sp1-dependent stimulation. The homeodomain is also required for cooperation of the AD with Sp1. By contrast, AD2 strongly activates transcription in an Sp1-independent manner only when the homeodomain has been removed. These observations indicate that HoxA-13 regulates transcription through multiple pathways. In addition, we found that a helix 3 mutation of the HoxA-13 homeodomain behaves as a dominant negative form.

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A mouse embryonic stem cell line showing pluripotency of differentiation in early embryos and ubiquitous β-galactosidase expression

Cited by 174 publications

References 18 publications

Keratinocyte-specific Expression of Fatty Acid Transport Protein 4 Rescues the Wrinkle-free Phenotype in Slc27a4/Fatp4 Mutant Mice

Keratinocyte-specific Expression of Fatty Acid Transport Protein 4 Rescues the Wrinkle-free Phenotype in Slc27a4/Fatp4 Mutant Mice

A Role for Receptor Protein Tyrosine Phosphatase in Midbrain Development

Hox Proteins Functionally Cooperate with the GC Box-binding Protein System through Distinct Domains

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