Advertisement
RESOURCE ARTICLE
New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
Ana Isabel Garcia Diaz, Ben Moyon, Philip M. Coan, Neza Alfazema, Lara Venda, Kevin Woollard, Tim Aitman
Disease Models & Mechanisms 2016 9: 463-471; doi: 10.1242/dmm.024208

Article Figures & Tables

Figures

  • Fig. 1.
    Fig. 1.

    GFP transgenic rat design. (A) Schematic plasmid representation. Rat elongation factor 1 alpha promoter (rEF1a) replaces CAG promoter (CAGGS). IR, inverted repeats; GFP, green fluorescent protein cDNA. (B) Photograph of WKY-GFP pups (left) and adult (right) rats under excitation light 489 nm, showing wild-type and GFP Tg animals. (C) Schematic genome locus showing TA integration sites location of transgene for SHR-GFP in chromosome 5, and WKY-GFP in chromosome 1 and 8. Genomic sequence (left junction) in capitals and transgene in lowercase, dotted horizontal line refers to intergenic, continuous black line to intronic, and vertical blocks to exonic sequences.

  • Fig. 2.
    Fig. 2.

    GFP expression in embryos. Female WKT-GFP rats were crossed with wild-type WKY male rats and one-cell embryos removed and imaged under confocal microscope. Representation of three experiments from one-cell embryos at E4.5 and E4.5 plus 12 h showing bright field (BF) images and expression of GFP. Scale bar: 15 μm (top panel); 40 μm (bottom panel).

  • Fig. 3.
    Fig. 3.

    GFP expression in organs and tissues. Wild type (from SHR strain), WKY-GFP and SHR-GFP rats were examined for gross GFP expression in dissected heart, brain, kidney, eyes, thymus, gut, liver, spleen and muscle tissues. Whole organ extracts were mounted for white light (left three panels) and stereo-fluorescence imaging (right three panels). Scale bar: 2 mm.

  • Fig. 4.
    Fig. 4.

    GFP expression in blood leukocyte populations. Anticoagulated blood from wild-type, WKY-GFP and SHR-GFP rats were examined for GFP expression with or without fluorescent-labelled antibodies using flow cytometry. (A) Histograms of GFP expression in SSChi and SSClow populations. (B) Lower panels: dot plots of GFP expression versus granulocyte (Gran)-, MHC class II (MHC-II)- or CD68-positive populations, in cells gated by R1 (upper panel). Representative of at least n=4 rats.

  • Fig. 5.
    Fig. 5.

    GFP expression in BMDM and intravital microscopy. (A) Bone marrow-derived macrophages (BMDM) were harvested and cultured for 5 or 10 days and intensity of GFP examined in WKY-GFP and SHR-GFP rats. Scale bar: 75 μm. (B). WKY rats underwent irradiation and bone marrow transplant from a WKY-GFP donor. After successful chimerisation, kidney cortex was imaged after injection of 70 kDa fluorescent dextran under anaesthesia. Snapshot is shown of GFP-positive cells interacting with endothelial surfaces within the kidney (arrows). Representative of at least n=4 rats. Scale bar: 40 μm.

Tables

  • Table 1.
Back to top

RSSRSS

Keywords

 Download PDF

Email
RESOURCE ARTICLE
New Wistar Kyoto and spontaneously hypertensive rat transgenic models with ubiquitous expression of green fluorescent protein
Ana Isabel Garcia Diaz, Ben Moyon, Philip M. Coan, Neza Alfazema, Lara Venda, Kevin Woollard, Tim Aitman
Disease Models & Mechanisms 2016 9: 463-471; doi: 10.1242/dmm.024208
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Subject collections

Other journals from The Company of Biologists

Advertisement

Editor’s choice – Perturbed development of cranial neural crest cells in association with reduced sonic hedgehog signaling underlies the pathogenesis of retinoic-acid-induced cleft palate

Schematic showing that excessive RA signaling reduces Shh signaling, which results in elevated cell death of CNCCs and cleft palate.

Takashi Yamashiro and colleagues identify the critical role of the retinoic acid-Sonic hedgehog signalling pathway in cranial neural crest and palate development.

A graphical 3D reconstruction of ventricular EDV and ESV as measured by CFIN.

A new Resource article by Nguyen et al. describes CFIN, a novel deep learning image analysis platform to assess cardiac function in embryonic zebrafish. 

Two pictures of non-human primates commonly used in genome editing research.

Yuyu Niu and colleagues summarise the history of genome editing in non-human primates and discusses the challenges and prospects of the technology.

Pictures of Alexander and Jocelyn.

Have you seen our interviews with the early-career first authors of our papers? Recently, we caught up with Alexander Akerberg and Jocelyn Wessels. 

Travelling Fellowship – New imaging approach unveils a bigger picture

Highlights from Travelling Fellowships trips.

Find out how Pamela Imperadore’s Travelling Fellowship grant from The Company of Biologists took her to Germany, where she used new imaging techniques to investigate the cellular machinery underlying octopus arm regeneration. Don’t miss the next application deadline for 2020 travel, coming up on 29 November. Where will you go?

preLights – HSP110 dependent HSP70 disaggregation machinery mediates prion-like propagation of amyloidogenic proteins in metazoa

preLights logo

Highlighted by Tessa Sinnige, a recent preprint from Carmen Nussbaum-Krammer and co-workers shows that Hsp110 activity protects against amorphous protein aggregation but promotes amyloid aggregation and toxicity in C. elegans models.