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cGMP-dependent protein kinase I is crucial for angiogenesis and postnatal vasculogenesis.

Aicher A, Heeschen C, Feil S, Hofmann F, Mendelsohn ME, Feil R, Dimmeler S - PLoS ONE (2009)

Bottom Line: Bone marrow progenitors from cGKI(-/-) mice showed reduced proliferation and survival rates.In addition, we used cGKIalpha leucine zipper mutant (LZM) mice as model for cGKI deficiency.Consistently, LZM mice exhibited reduced numbers of vasculogenic progenitors and impaired neovascularization following hindlimb ischemia compared to WT mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine III, JW Goethe University, Frankfurt, Germany. aicher_a@yahoo.com

ABSTRACT

Background: Endothelium-derived nitric oxide plays an important role for the bone marrow microenvironment. Since several important effects of nitric oxide are mediated by cGMP-dependent pathways, we investigated the role of the cGMP downstream effector cGMP-dependent protein kinase I (cGKI) on postnatal neovascularization.

Methodology/principal findings: In a disc neovascularization model, cGKI(-/-) mice showed an impaired neovascularization as compared to their wild-type (WT) littermates. Infusion of WT, but not cGKI(-/-) bone marrow progenitors rescued the impaired ingrowth of new vessels in cGKI-deficient mice. Bone marrow progenitors from cGKI(-/-) mice showed reduced proliferation and survival rates. In addition, we used cGKIalpha leucine zipper mutant (LZM) mice as model for cGKI deficiency. LZM mice harbor a mutation in the cGKIalpha leucine zipper that prevents interaction with downstream signaling molecules. Consistently, LZM mice exhibited reduced numbers of vasculogenic progenitors and impaired neovascularization following hindlimb ischemia compared to WT mice.

Conclusions/significance: Our findings demonstrate that the cGMP-cGKI pathway is critical for postnatal neovascularization and establish a new role for cGKI in vasculogenesis, which is mediated by bone marrow-derived progenitors.

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Functional capacity of BMC from cGKI−/− and WT mice.(A) Representative FACS analysis of Lin− BMC after pulsing with BrdU for 1 h and staining with BrdU-FITC and sca-1-PE. (B) Proliferation (BrdU+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 4). (C) Apoptosis (annexinV+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 3).
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pone-0004879-g004: Functional capacity of BMC from cGKI−/− and WT mice.(A) Representative FACS analysis of Lin− BMC after pulsing with BrdU for 1 h and staining with BrdU-FITC and sca-1-PE. (B) Proliferation (BrdU+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 4). (C) Apoptosis (annexinV+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 3).

Mentions: To provide further insights into the regulation of vasculogenesis through cGMP-dependent pathways, we examined the role of cGKI on the functional capacity of bone marrow-derived progenitor cells. First, we demonstrated that cGKI is expressed in freshly isolated CD45+ hematopoietic bone marrow cells, freshly isolated total bone marrow, as well as cultured bone marrow stromal cells (Fig. 3A+B). The most prominent expression of cGKI was found in cultured bone marrow stromal cells, most likely due the relative enrichment of endothelial cells and fibroblasts in cultured bone marrow stromal cells. To assess the functional capacity of bone marrow-derived progenitor cells, we measured proliferation and apoptosis rates in Lin−sca-1+ bone marrow progenitor cells (Fig. 4A–C). BrdU incorporation as a marker of cell proliferation was assessed by flow cytometry and demonstrated a strong reduction in Lin−sca-1+ bone marrow progenitor cells from cGKI−/− mice (Fig. 4A+B). Simultaneously, the number of apoptotic Lin−sca-1+ bone marrow progenitor cells was increased in cGKI−/− mice as assessed by annexin V binding (Fig. 4C). These results suggest that cGKI promotes proliferation and survival of bone marrow-derived progenitor cells, which contributes to neovascularization.


cGMP-dependent protein kinase I is crucial for angiogenesis and postnatal vasculogenesis.

Aicher A, Heeschen C, Feil S, Hofmann F, Mendelsohn ME, Feil R, Dimmeler S - PLoS ONE (2009)

Functional capacity of BMC from cGKI−/− and WT mice.(A) Representative FACS analysis of Lin− BMC after pulsing with BrdU for 1 h and staining with BrdU-FITC and sca-1-PE. (B) Proliferation (BrdU+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 4). (C) Apoptosis (annexinV+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 3).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654072&req=5

pone-0004879-g004: Functional capacity of BMC from cGKI−/− and WT mice.(A) Representative FACS analysis of Lin− BMC after pulsing with BrdU for 1 h and staining with BrdU-FITC and sca-1-PE. (B) Proliferation (BrdU+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 4). (C) Apoptosis (annexinV+) of sca-1+Lin− bone marrow progenitor cells from cGKI−/− and WT mice (gated on lymphocyte-monocyte fraction; n = 3).
Mentions: To provide further insights into the regulation of vasculogenesis through cGMP-dependent pathways, we examined the role of cGKI on the functional capacity of bone marrow-derived progenitor cells. First, we demonstrated that cGKI is expressed in freshly isolated CD45+ hematopoietic bone marrow cells, freshly isolated total bone marrow, as well as cultured bone marrow stromal cells (Fig. 3A+B). The most prominent expression of cGKI was found in cultured bone marrow stromal cells, most likely due the relative enrichment of endothelial cells and fibroblasts in cultured bone marrow stromal cells. To assess the functional capacity of bone marrow-derived progenitor cells, we measured proliferation and apoptosis rates in Lin−sca-1+ bone marrow progenitor cells (Fig. 4A–C). BrdU incorporation as a marker of cell proliferation was assessed by flow cytometry and demonstrated a strong reduction in Lin−sca-1+ bone marrow progenitor cells from cGKI−/− mice (Fig. 4A+B). Simultaneously, the number of apoptotic Lin−sca-1+ bone marrow progenitor cells was increased in cGKI−/− mice as assessed by annexin V binding (Fig. 4C). These results suggest that cGKI promotes proliferation and survival of bone marrow-derived progenitor cells, which contributes to neovascularization.

Bottom Line: Bone marrow progenitors from cGKI(-/-) mice showed reduced proliferation and survival rates.In addition, we used cGKIalpha leucine zipper mutant (LZM) mice as model for cGKI deficiency.Consistently, LZM mice exhibited reduced numbers of vasculogenic progenitors and impaired neovascularization following hindlimb ischemia compared to WT mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine III, JW Goethe University, Frankfurt, Germany. aicher_a@yahoo.com

ABSTRACT

Background: Endothelium-derived nitric oxide plays an important role for the bone marrow microenvironment. Since several important effects of nitric oxide are mediated by cGMP-dependent pathways, we investigated the role of the cGMP downstream effector cGMP-dependent protein kinase I (cGKI) on postnatal neovascularization.

Methodology/principal findings: In a disc neovascularization model, cGKI(-/-) mice showed an impaired neovascularization as compared to their wild-type (WT) littermates. Infusion of WT, but not cGKI(-/-) bone marrow progenitors rescued the impaired ingrowth of new vessels in cGKI-deficient mice. Bone marrow progenitors from cGKI(-/-) mice showed reduced proliferation and survival rates. In addition, we used cGKIalpha leucine zipper mutant (LZM) mice as model for cGKI deficiency. LZM mice harbor a mutation in the cGKIalpha leucine zipper that prevents interaction with downstream signaling molecules. Consistently, LZM mice exhibited reduced numbers of vasculogenic progenitors and impaired neovascularization following hindlimb ischemia compared to WT mice.

Conclusions/significance: Our findings demonstrate that the cGMP-cGKI pathway is critical for postnatal neovascularization and establish a new role for cGKI in vasculogenesis, which is mediated by bone marrow-derived progenitors.

Show MeSH
Related in: MedlinePlus