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Matrix metalloproteinase-2, caveolins, focal adhesion kinase and c-Kit in cells of the mouse myocardium.

Cho WJ, Chow AK, Schulz R, Daniel EE - J. Cell. Mol. Med. (2007 Sep-Oct)

Bottom Line: In Cav-1(-/-) LV Cav-1 and MMP-2 were absent or reduced everywhere.To conclude, MMP-2 is closely associated with caveolins at FB and CEC as well as at CM.Also, MMP-2 is closely associated with FAK at FB and c-Kit at ICLC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.

ABSTRACT
Matrix metalloproteinase-2 (MMP-2) may play roles at intracellular and extracellular sites of the heart in ischaemia/reperfusion injury. Caveolins (Cav-1, -2 and -3) are lipid raft proteins which play roles in cell sig-nalling. This study examined, using immunohistochemistry and two photon confocal microscopy, if MMP-2 and caveolins co-localize at the plasma membrane of cardiac cells: cardiomyocytes (CM), fibroblasts (FB) and capillary endothelial cells (CEC) in the left ventricle (LV) of the Cav-1(+/+) and Cav-1(-/-) mouse heart. In Cav-1(+/+) mouse LV MMP-2 and Cav-1 co-localized at CM plasma membranes, and at multiple locations in FB and CEC. MMP-2 co-localized with Cav-2 only at CEC. MMP-2 co-localized with Cav-3 at CM plasma membranes and Z-lines, and partially at FB and CEC. In Cav-1(-/-) LV Cav-1 and MMP-2 were absent or reduced everywhere. Cav-2 appeared at CEC despite the absence of Cav-1. Cav-3 appeared at CM plasma membranes and Z-lines, FB and CEC. Also, FAK in FB and c-Kit in interstitial Cajal-like cells (ICLC) were completely absent. By transmission electron microscopy in Cav-1(+/+), regular size caveolae (Cav) were at CEC, irregular size Cav were at CM and a few were at FB. In Cav-1(-/-) there were few Cav at CM and FB and some at CEC. To conclude, MMP-2 is closely associated with caveolins at FB and CEC as well as at CM. Also, MMP-2 is closely associated with FAK at FB and c-Kit at ICLC. Thus, Cav-1 expression is not necessary for Cav-2 expression. Cav-3 or Cav-3 with Cav-2 has the capability to make Cav.

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Co-localization of Cav-3 and FAK / DDR-2 / vWF / c-Kit in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ mouse some FB are Cav-3- (green) and FAK-positive (red) and the two proteins are partially co-localized (yellow) (large arrows), besides some FB are Cav-3-positive but FAK-negative (small arrows) (A–C). Some Cav-3-positive FB (red) are DDR-2-positive (green) and the two proteins are partially co-localized in FB tails (arrows) (E–G). vWF-positive CEC (arrows) do not have Cav-3 (I–K). ICLC are both Cav-3- (green) and c-Kit-positive (red) and the two proteins are partially co-localized (yellow) (M–O). In Cav-1−/− mouse Cav-3 appeared at CM plasma membranes, Z-lines and FB (arrows), but FAK and c-Kit are completely absent (D and P). DDR-2 and vWF shows at FB and CEC, respectively (H and I). Scale bar is 10 μm for a–d and i–p, and 7 μm for e–g.
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fig06: Co-localization of Cav-3 and FAK / DDR-2 / vWF / c-Kit in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ mouse some FB are Cav-3- (green) and FAK-positive (red) and the two proteins are partially co-localized (yellow) (large arrows), besides some FB are Cav-3-positive but FAK-negative (small arrows) (A–C). Some Cav-3-positive FB (red) are DDR-2-positive (green) and the two proteins are partially co-localized in FB tails (arrows) (E–G). vWF-positive CEC (arrows) do not have Cav-3 (I–K). ICLC are both Cav-3- (green) and c-Kit-positive (red) and the two proteins are partially co-localized (yellow) (M–O). In Cav-1−/− mouse Cav-3 appeared at CM plasma membranes, Z-lines and FB (arrows), but FAK and c-Kit are completely absent (D and P). DDR-2 and vWF shows at FB and CEC, respectively (H and I). Scale bar is 10 μm for a–d and i–p, and 7 μm for e–g.

Mentions: Both FAK-FB and FAK-negative cells were present in the interstitial space. Only a few of FAK-FBs also contained Cav-3, which was co-localized with FAK. The FAK-negative cells were all Cav-3-positive cells. The FAK-negative Cav-3-positive cells might be DDR-2-FB or c-Kit-FB in Cav-1+/+ mouse LVM (Fig. 6A–C). FAK was absent in non-nuclear portions of cells but Cav-3 appeared at FB in interstitial space in Cav-1−/− mouse LVM (Fig. 6D).


Matrix metalloproteinase-2, caveolins, focal adhesion kinase and c-Kit in cells of the mouse myocardium.

Cho WJ, Chow AK, Schulz R, Daniel EE - J. Cell. Mol. Med. (2007 Sep-Oct)

Co-localization of Cav-3 and FAK / DDR-2 / vWF / c-Kit in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ mouse some FB are Cav-3- (green) and FAK-positive (red) and the two proteins are partially co-localized (yellow) (large arrows), besides some FB are Cav-3-positive but FAK-negative (small arrows) (A–C). Some Cav-3-positive FB (red) are DDR-2-positive (green) and the two proteins are partially co-localized in FB tails (arrows) (E–G). vWF-positive CEC (arrows) do not have Cav-3 (I–K). ICLC are both Cav-3- (green) and c-Kit-positive (red) and the two proteins are partially co-localized (yellow) (M–O). In Cav-1−/− mouse Cav-3 appeared at CM plasma membranes, Z-lines and FB (arrows), but FAK and c-Kit are completely absent (D and P). DDR-2 and vWF shows at FB and CEC, respectively (H and I). Scale bar is 10 μm for a–d and i–p, and 7 μm for e–g.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401273&req=5

fig06: Co-localization of Cav-3 and FAK / DDR-2 / vWF / c-Kit in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ mouse some FB are Cav-3- (green) and FAK-positive (red) and the two proteins are partially co-localized (yellow) (large arrows), besides some FB are Cav-3-positive but FAK-negative (small arrows) (A–C). Some Cav-3-positive FB (red) are DDR-2-positive (green) and the two proteins are partially co-localized in FB tails (arrows) (E–G). vWF-positive CEC (arrows) do not have Cav-3 (I–K). ICLC are both Cav-3- (green) and c-Kit-positive (red) and the two proteins are partially co-localized (yellow) (M–O). In Cav-1−/− mouse Cav-3 appeared at CM plasma membranes, Z-lines and FB (arrows), but FAK and c-Kit are completely absent (D and P). DDR-2 and vWF shows at FB and CEC, respectively (H and I). Scale bar is 10 μm for a–d and i–p, and 7 μm for e–g.
Mentions: Both FAK-FB and FAK-negative cells were present in the interstitial space. Only a few of FAK-FBs also contained Cav-3, which was co-localized with FAK. The FAK-negative cells were all Cav-3-positive cells. The FAK-negative Cav-3-positive cells might be DDR-2-FB or c-Kit-FB in Cav-1+/+ mouse LVM (Fig. 6A–C). FAK was absent in non-nuclear portions of cells but Cav-3 appeared at FB in interstitial space in Cav-1−/− mouse LVM (Fig. 6D).

Bottom Line: In Cav-1(-/-) LV Cav-1 and MMP-2 were absent or reduced everywhere.To conclude, MMP-2 is closely associated with caveolins at FB and CEC as well as at CM.Also, MMP-2 is closely associated with FAK at FB and c-Kit at ICLC.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada.

ABSTRACT
Matrix metalloproteinase-2 (MMP-2) may play roles at intracellular and extracellular sites of the heart in ischaemia/reperfusion injury. Caveolins (Cav-1, -2 and -3) are lipid raft proteins which play roles in cell sig-nalling. This study examined, using immunohistochemistry and two photon confocal microscopy, if MMP-2 and caveolins co-localize at the plasma membrane of cardiac cells: cardiomyocytes (CM), fibroblasts (FB) and capillary endothelial cells (CEC) in the left ventricle (LV) of the Cav-1(+/+) and Cav-1(-/-) mouse heart. In Cav-1(+/+) mouse LV MMP-2 and Cav-1 co-localized at CM plasma membranes, and at multiple locations in FB and CEC. MMP-2 co-localized with Cav-2 only at CEC. MMP-2 co-localized with Cav-3 at CM plasma membranes and Z-lines, and partially at FB and CEC. In Cav-1(-/-) LV Cav-1 and MMP-2 were absent or reduced everywhere. Cav-2 appeared at CEC despite the absence of Cav-1. Cav-3 appeared at CM plasma membranes and Z-lines, FB and CEC. Also, FAK in FB and c-Kit in interstitial Cajal-like cells (ICLC) were completely absent. By transmission electron microscopy in Cav-1(+/+), regular size caveolae (Cav) were at CEC, irregular size Cav were at CM and a few were at FB. In Cav-1(-/-) there were few Cav at CM and FB and some at CEC. To conclude, MMP-2 is closely associated with caveolins at FB and CEC as well as at CM. Also, MMP-2 is closely associated with FAK at FB and c-Kit at ICLC. Thus, Cav-1 expression is not necessary for Cav-2 expression. Cav-3 or Cav-3 with Cav-2 has the capability to make Cav.

Show MeSH
Related in: MedlinePlus