Blog
About

8
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Long-Range Control of Renin Gene Expression in Tsukuba Hypertensive Mice

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Renin, a rate-limiting enzyme in the renin–angiotensin system, is regulated to maintain blood pressure homeostasis: renin gene expression in the kidney is suppressed in a hypertensive environment. We found that expression of a 15-kb human RENIN (h REN) transgene was aberrantly upregulated (>4.2-fold), while the endogenous mouse renin (m Ren) gene was suppressed (>1.7-fold) in Tsukuba hypertensive mice (THM), a model for genetically induced hypertension. We then generated transgenic mice using a 13-kb m Ren gene fragment that was homologous to the 15-kb h REN transgene and found that its expression was also upregulated (>3.1-fold) in THM, suggesting that putative silencing elements of the renin genes were distally located in the loci. We next examined the possible role of a previously identified mouse distal enhancer (mdE) located outside of the 13-kb m Ren gene fragment. Deletion of the mdE in the context of a 156-kb m Ren transgene did not affect its transcriptional repression in THM, implying that although the silencing element of the m Ren gene is located within the 156-kb fragment tested, it is distinct from the mdE. Consistent with these results, deletion of the 63-kb region upstream of the mdE from the endogenous m Ren gene locus abrogated its transcriptional repression in THM. We finally tested whether dysregulation of the short renin transgenes also occurred in the fetal or neonatal kidneys of THM and found that their expression was not aberrantly upregulated, demonstrating that aberrant regulation of short renin transgenes commences sometime between neonate and adult periods.

          Related collections

          Most cited references 34

          • Record: found
          • Abstract: found
          • Article: not found

          A highly efficient Escherichia coli-based chromosome engineering system adapted for recombinogenic targeting and subcloning of BAC DNA.

          Recently, a highly efficient recombination system for chromosome engineering in Escherichia coli was described that uses a defective lambda prophage to supply functions that protect and recombine a linear DNA targeting cassette with its substrate sequence (Yu et al., 2000, Proc. Natl. Acad. Sci. USA 97, 5978-5983). Importantly, the recombination is proficient with DNA homologies as short as 30-50 bp, making it possible to use PCR-amplified fragments as the targeting cassette. Here, we adapt this prophage system for use in bacterial artificial chromosome (BAC) engineering by transferring it to DH10B cells, a BAC host strain. In addition, arabinose inducible cre and flpe genes are introduced into these cells to facilitate BAC modification using loxP and FRT sites. Next, we demonstrate the utility of this recombination system by using it to target cre to the 3' end of the mouse neuron-specific enolase (Eno2) gene carried on a 250-kb BAC, which made it possible to generate BAC transgenic mice that specifically express Cre in all mature neurons. In addition, we show that fragments as large as 80 kb can be subcloned from BACs by gap repair using this recombination system, obviating the need for restriction enzymes or DNA ligases. Finally, we show that BACs can be modified with this recombination system in the absence of drug selection. The ability to modify or subclone large fragments of genomic DNA with precision should facilitate many kinds of genomic experiments that were difficult or impossible to perform previously and aid in studies of gene function in the postgenomic era. Copyright 2001 Academic Press.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Cyclooxygenase-2 is associated with the macula densa of rat kidney and increases with salt restriction.

            The kidney is a rich source of prostaglandins. These eicosanoids, formed by cyclooxygenase-dependent metabolism of arachidonic acid, are important physiologic mediators of renal glomerular hemodynamics and tubular sodium and water reabsorption. Two separate isoforms of cyclooxygenase (COX) have now been identified: constitutive COX-1, encoded by a 2.8-kb mRNA, and mitogen-activated COX-2, encoded by a 4.0-4.5-kb mRNA. COX-2 expression increases during development and inflammation, but, except for brain, constitutive expression is low. It has been generally accepted that physiologic renal production of prostaglandins is mediated by COX-1. However, in the absence of inflammation, low levels of COX-2 mRNA are also detectable in the kidney. To examine the role of COX-2 in the kidney and determine its intrarenal localization, we used a 1.3-kb cDNA probe specific for the 3' untranslated region of rat COX-2 and COX-2-specific antiserum. The COX-2-specific cDNA probe hybridized with a 4.4-kb transcript in total RNA from adult rat kidney. Immunoblots of microsomes isolated from kidney cortex and papilla indicated immunoreactive COX-2 in both locations. In situ hybridization and immunohistochemistry indicated that renal cortical COX-2 expression was localized to the macula densa of the juxtaglomerular apparatus and to adjacent epithelial cells of the cortical thick ascending limb of Henle. In addition, COX-2 immunoreactivity was detected in interstitial cells in the papilla. No COX-2 message or immunoreactive protein was detected in arterioles, glomeruli, or cortical or medullary collecting ducts. When animals were chronically sodium restricted, the level of COX-2 in the region of the macula densa increased threefold (from 0.86 +/- 0.08 to 2.52 +/- 0.43/mm2) and the total area of the COX-2 immunoreactive cells in cortex increased from 34 microns2/mm2 of cortex to 226 microns2/mm2 of cortex. The intrarenal distribution of COX-2 and its increased expression in response to sodium restriction suggest that in addition to its proposed role in inflammatory and growth responses, this enzyme may play an important role in the regulation of salt, volume, and blood pressure homeostasis.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Genome organization and long-range regulation of gene expression by enhancers.

              It is now well accepted that cell-type specific gene regulation is under the purview of enhancers. Great strides have been made recently to characterize and identify enhancers both genetically and epigenetically for multiple cell types and species, but efforts have just begun to link enhancers to their target promoters. Mapping these interactions and understanding how the 3D landscape of the genome constrains such interactions is fundamental to our understanding of mammalian gene regulation. Here, we review recent progress in mapping long-range regulatory interactions in mammalian genomes, focusing on transcriptional enhancers and chromatin organization principles. Copyright © 2013. Published by Elsevier Ltd.
                Bookmark

                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                18 November 2016
                2016
                : 11
                : 11
                Affiliations
                [1 ]Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan
                [2 ]Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan
                [3 ]Life Science Center, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Ibaraki, Japan
                Max Delbruck Centrum fur Molekulare Medizin Berlin Buch, GERMANY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: AU KT.

                • Data curation: AU.

                • Formal analysis: AU JI.

                • Funding acquisition: AF KT.

                • Investigation: AU HM KT.

                • Methodology: AU AF KT.

                • Project administration: AF KT.

                • Resources: JI AF.

                • Validation: JI.

                • Writing – original draft: AU KT.

                Article
                PONE-D-16-36683
                10.1371/journal.pone.0166974
                5115840
                27861631
                © 2016 Ushiki et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 8, Tables: 2, Pages: 20
                Product
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100007263, Astellas Foundation for Research on Metabolic Disorders;
                Award Recipient :
                Funded by: Japan Heart Foundation
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100008732, Uehara Memorial Foundation;
                Award Recipient :
                Funded by: Tokyo Biochemical Research Foundation
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100007449, Takeda Science Foundation;
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001691, Japan Society for the Promotion of Science;
                Award ID: KAKENHI Grant Number 16J04305
                Award Recipient :
                This study was supported by The Astellas Foundation for Research on Metabolic Disorders ( https://www.astellas.com/jp/byoutai/) (KT); The Japan Heart Foundation ( http://www.jhf.or.jp/english/) (KT); The Uehara Memorial Foundation ( http://www.ueharazaidan.or.jp) (KT); The Tokyo Biochemical Research Foundation ( http://www.tokyobrf.or.jp/english/) (KT); The Takeda Science Foundation ( http://www.takeda-sci.or.jp) (KT); The Japan Society for the Promotion of Science (JSPS KAKENHI grant number 16J04305) ( https://www.jsps.go.jp/english/) (AU). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Renal System
                Kidneys
                Medicine and Health Sciences
                Anatomy
                Renal System
                Kidneys
                Biology and life sciences
                Genetics
                Gene expression
                DNA transcription
                Biology and Life Sciences
                Genetics
                Gene Expression
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Medicine and Health Sciences
                Vascular Medicine
                Blood Pressure
                Hypertension
                Medicine and Health Sciences
                Vascular Medicine
                Blood Pressure
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Silencer Elements
                Research and Analysis Methods
                Model Organisms
                Animal Models
                Mouse Models
                Custom metadata
                All relevant data are within the paper.

                Uncategorized

                Comments

                Comment on this article