In vivo endothelial gene regulation in diabetes

Maresh, J. Gregory; Shohet, Ralph V.
January 2008
Cardiovascular Diabetology;2008, Vol. 7, p1
Academic Journal
Background: An authentic survey of the transcript-level response of the diabetic endothelium in vivo is key to understanding diabetic cardiovascular complications such as accelerated atherosclerosis and endothelial dysfunction. Methods: We used streptozotocin to induce a model of type I diabetes in transgenic mice that express green fluorescent protein under the control of an endothelial-specific promoter (Tie2-GFP) allowing rapid isolation of aortic endothelium. Three weeks after treatment, endothelial cells were isolated from animals with blood glucose > 350 mg/dl. Aortae from the root to the renal bifurcation were rapidly processed by mincing and proteolytic digestion followed by fluorescent activated cell sorting to yield endothelial cell populations of >95% purity. RNA was isolated from >50,000 endothelial cells and subjected to oligo dT amplification prior to transcriptional analysis on microarrays displaying long oligonucleotides representing 32,000 murine transcripts. Five regulated transcripts were selected for analysis by real-time PCR. Results: Within replicate microarray experiments, 19 transcripts were apparently dysregulated by at least 70% within diabetic mice. Up-regulation of glycam1, slc36a2, ces3, adipsin and adiponectin was confirmed by real-time PCR. Conclusion: By comprehensively examining cellular gene responses in vivo in a whole animal model of type I diabetes, we have identified novel regulation of key endothelial transcripts that likely contribute to the metabolic and pro-inflammatory responses that accompany diabetes.


Related Articles

  • Cell-penetrating peptide (CPP)-conjugated proteins is an efficient tool for manipulation of human mesenchymal stromal cells. Junghyun Jo; Soomin Hong; Won Yun Choi; Dong Ryul Lee // Scientific Reports;2/14/2014, p1 

    Delivery of proteins has been regarded as the safest and most useful application in therapeutic application of stem cells, because proteins can regulate gene expression transiently without any genomic alteration. However, it is difficult to accurately measure efficiency or quantity of...

  • Gene network shaping of inherent noise spectra. Austin, D. W.; Allen, M. S.; McCollum, J. M.; Dar, R. D.; Wilgus, J. R.; Sayler, G. S.; Samatova, N. F.; Cox, C. D.; Simpson, M. L. // Nature;2/2/2006, Vol. 439 Issue 7076, p608 

    Recent work demonstrates that stochastic fluctuations in molecular populations have consequences for gene regulation. Previous experiments focused on noise sources or noise propagation through gene networks by measuring noise magnitudes. However, in theoretical analysis, we showed that noise...

  • Hepatic stellate cell-specific gene silencing induced by an artificial microRNA for antifibrosis in vitro. Ying Chang; Hua-jun Jiang; Xue-mei Sun; Xiao-kun Cai; Xing-xing He; Pei-yuan Li; Wang-xian Tang; Yu-hu Song; Ju-sheng Lin; Chang, Ying; Jiang, Hua-jun; Sun, Xue-mei; Cai, Xiao-kun; He, Xing-xing; Li, Pei-yuan; Tang, Wang-xian; Song, Yu-hu; Lin, Ju-sheng // Digestive Diseases & Sciences;Mar2010, Vol. 55 Issue 3, p642 

    Background: We previously reported that the anti-transforming growth factor-beta1 (TGF-beta1) ribozymes directed by T7 and CMV promoters could reverse the character of activated hepatic stellate cells (HSCs) in vitro and improve fibrotic pathology in vivo. However, nonspecific...

  • GFP expressing bacterial biosensor to measure lead contamination in aquatic environment. Chakraborty, Tapas; Babu, P. Gireesh; Alam, Absar; Chaudhari, Aparna // Current Science (00113891);3/25/2008, Vol. 94 Issue 6, p800 

    A bacterial biosensor is reported that responds to Pb2+ in the range of 50-400 µM by expressing green fluorescence protein (GFP). The genetic element that senses Pb2+ includes the regulatory protein gene (PbrR) along with operator/promoter (PbrO/P) of the lead resistance operon from plasmid...

  • Localization and Mobility of Synaptic Vesicles in Myosin VI Mutants of Drosophila. Kisiel, Marta; McKenzie, Kristopher; Stewart, Bryan // PLoS ONE;Jul2014, Vol. 9 Issue 7, p1 

    Background: At the Drosophila neuromuscular junction (NMJ), synaptic vesicles are mobile; however, the mechanisms that regulate vesicle traffic at the nerve terminal are not fully understood. Myosin VI has been shown to be important for proper synaptic physiology and morphology at the NMJ,...

  • Protection against Autoimmune Diabetes by Silkworm-Produced GFP-Tagged CTB-Insulin Fusion Protein. Qiaohong Meng; Wenfeng Wang; Xiaowen Shi; Yongfeng Jin; Yaozhou Zhang // Clinical & Developmental Immunology;2011, p1 

    In animals, oral administration of the cholera toxin B (CTB) subunit conjugated to the autoantigen insulin enhances the specific immune-unresponsive state. This is called oral tolerance and is capable of suppressing autoimmune type 1 diabetes (T1D). However, the process by which the CTB-insulin...

  • Retinoic Acid Induces Pdx1-Positive Endoderm in Differentiating Mouse Embryonic Stem Cells. Micallef, Suzanne J.; Janes, Mary E.; Knezevic, Kathy; Davis, Richard P.; Elefanty, Andrew G.; Stanley, Edouard G. // Diabetes;Feb2005, Vol. 54 Issue 2, p301 

    We have generated an embryonic stem (ES) cell line in which sequences encoding green fluorescent protein (GFP) were targeted to the locus of the pancreatic-duodenal homeobox gene (Pdx1). Analysis of chimeric embryos derived from blastocyst injection of Pdx1GFP/w ES cells demonstrated that the...

  • Ferritin as an Endogenous MRI Reporter for Noninvasive Imaging of Gene Expression in C6 Glioma Tumors. Cohen, Batya; Dafni, Hagit; Meir, Gila; Neeman, Michal; Harmelin, Alon // Neoplasia;Feb2005, Vol. 7 Issue 2, p109 

    The heavy chain of murine ferritin, an iron storage molecule with ferroxidase activity, was developed as a novel endogenous reporter for the detection of gene expression by magnetic resonance imaging (MRI). Expression of both enhanced green fluorescent protein (EGFP) and influenza hemagglutinin...

  • Tom1l2 hypomorphic mice exhibit increased incidence of infections and tumors and abnormal immunologic response. Girirajan, Santhosh; Hauck, Paula; Williams, Stephen; Vlangos, Christopher; Szomju, Barbara; Solaymani-Kohal, Sara; Mosier, Philip; White, Kimber; McCoy, Kathleen; Elsea, Sarah // Mammalian Genome;Apr2008, Vol. 19 Issue 4, p246 

    Studies have shown that the TOM1 family of proteins, including TOM1 and TOM1L1, are actively involved in endosomal trafficking and function in the immune response. However, much less is known about the function of TOM1L2. To understand the biological importance of TOM1L2 and the potential...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics