Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Feb;101(2):e48-51.
doi: 10.3324/haematol.2015.137026.

Iron deficiency anemia in cyclic GMP kinase knockout mice

Elisabeth Angermeier  1 , Katrin Domes  1 , Robert Lukowski  2 , Jens Schlossmann  3 , Birgit Rathkolb  4 , Martin Hraběde Angelis  5 , Franz Hofmann  6
Affiliations

Iron deficiency anemia in cyclic GMP kinase knockout mice

Elisabeth Angermeier et al. Haematologica. 2016 Feb.
No abstract available

Keywords: H+K+-ATPase; cyclic GMP (cGMP); erythrocyte; gene knockout; hemoglobin; protein kinase G (PKG).

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Western blot of cGKI in WT Erythrocytes. (A,B,C) cGKI Western blots were performed in three different laboratories using different methods. Details of preparation of cells are given in Online Supplementary Methods. (A) 30 (platelets) and 40 (erythrocytes) μg protein lysates were loaded per slot. Proteins of slots 1 and 5, 2 and 6, 3 and 7, and 4 and 8 were from the same mouse preparation. A transferrin receptor 1 (TfRc) specific antibody was used to identify the erythrocytes. Equal loading of the gel was demonstrated by detecting β-actin. The cGMP-degrading phosphodiesterase 5 (PDE-5) was only present in platelets allowing for further differentiation between erythrocyte and platelet lysates. (B) Erythrocytes were purified by differential centrifugation from wild-type and cGKIα RM mice. 70 μg protein was loaded per slot. As control, 70 μg protein from WT stomach were analyzed with antibodies directed against cGKI, inositol 1,4,5-trisphosphate receptor associated cGMP kinase substrate (IRAG) or GAPDH. (C) 45 μg protein from erythrocytes purified by immunomagnetic selection using magnetically labeled Anti-Ter119 MicroBeads were loaded per slot. 100 μg WT spleen proteins were loaded per slot. In Figure 1Ca 5 ng purified cGKIα was loaded on the slot. For better comparison, Western blots were developed for 4 sec (Figure 1Ca), 30 sec (Figure 1Cb), and 1 sec (Figure 1Cc). Hbα: hemoglobin α-chain.
Figure 2.
Figure 2.
Effect of PPI on erythrocytes, hemoglobin, reticulocytes, mean corpuscular volume, and red blood cell distribution width (RDW-CV). Blood was taken from individual 6 to 12 week old mice. The number of mice is given within each column. Mice were not treated (−PPI) or treated with PPI (+PPI). A) erythrocytes; B) hemoglobin; C) hematocrit; D) reticulocytes; E) mean corpuscular volume (MCV); F) red blood cell distribution width (RDW-CV). ***P<0.001; **P<0.01; *P<0.05.
Figure 3.
Figure 3.
Plasma iron concentration (A), spleen weight (B), and quantitative real-time PCR of liver HAMP (C) and transferrin receptor 1 (D) mRNA. A) Plasma iron and B) spleen weight were measured before (−PPI) and after (+PPI) treatment with PPI. The number of mice is given below each scatter plot. C) mRNA of liver hepcidin. cGKI−/− mice were injected i.p. with physiological sodium chloride (NaCl-Inj.) or with Fe3+ for 12 days (see Online Supplementary Methods). WT mice were not treated with PPI. D) mRNA of liver transferrin receptor 1 (TfRc). mRNA was extracted from the liver of WT and cGKI−/− (KO) mice. mRNA levels for hepcidin and transferrin receptor 1 were normalized to mRNA levels for GAPDH. Mice were not treated (−PPI) or treated (+PPI) with PPI. The number of mice is given within the columns. ***P<0.001; **P<0.01; *P<0.05.
See this image and copyright information in PMC

References

    1. Weber S, Bernhard D, Lukowski R, et al. Rescue of cGMP kinase I knockout mice by smooth muscle specific expression of either isozyme. Circ Res. 2007;101(11):1096–1103. - PubMed
    1. Singh AK, Spiessberger B, Zheng W, et al. Neuronal cGMP kinase I is essential for stimulation of duodenal bicarbonate secretion by luminal acid. FASEB J. 2012;26(4):1745–1754. - PubMed
    1. Foller M, Feil S, Ghoreschi K, et al. Anemia and splenomegaly in cGKI-deficient mice. Proc Natl Acad Sci USA. 2008;105(18):6771–6776. - PMC - PubMed
    1. Zhang L, Lukowski R, Gaertner F, et al. Thrombocytosis as a response to high interleukin-6 levels in cGMP-dependent protein kinase I mutant mice. Arterioscler Tromb Vasc Biol. 2013;33(8):1820–1828. - PubMed
    1. Lang F, Abed M, Lang E, Foller M. Oxidative stress and suicidal erythrocyte death. AntioxidRedox Signal. 2014;21(1):138–153. - PubMed

Publication types

MeSH terms