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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 between caveolins in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ 3D reconstructed images (-45° rotation of y-axis) shows Cav-1 (red) localizes at both FB (nucleus: large asterisks) and CEC (nucleus: small asterisks), but Cav-2 (green) localizes only at CEC. Partial co-localization of Cav-1 and Cav-2 occurs at CEC (open arrows), but sometimes Cav-1 without Cav-2 occurs at CEC membrane (closed arrows) (A–C). Cav-1 (red) co-localizes well with Cav-3 (green) at CM plasma membrane (large open arrow), FB (nucleus: large asterisks) interconnecting with tails (small open arrow) and CEC (nucleus: small asterisks). Also, Cav-1 immunoreactivity occurs near CM Z-lines and partial co-localization with Cav-3 (E–G). Cav-2 (green) co-localizes partially with Cav-3 (red) at CEC (arrows). CEC make frequently anastomotic structure (asterisk) (I–K). In Cav-1−/− Cav-2 (green) immunore-activity occurs regardless of Cav-1 at CEC (d). Cav-3 (green), immunoreactivity occurs also at CM plasma membranes, Z-lines and FB (asterisks) (H). Co-localization (open arrows) of Cav-2 (green) and Cav-3 (red) at CEC is similar to that in Cav-1+/+(I). CM is cardiomyocyte and IS is interstitial space. Scale bar is 10 mm for all images.
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fig03: Co-localization between caveolins in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ 3D reconstructed images (-45° rotation of y-axis) shows Cav-1 (red) localizes at both FB (nucleus: large asterisks) and CEC (nucleus: small asterisks), but Cav-2 (green) localizes only at CEC. Partial co-localization of Cav-1 and Cav-2 occurs at CEC (open arrows), but sometimes Cav-1 without Cav-2 occurs at CEC membrane (closed arrows) (A–C). Cav-1 (red) co-localizes well with Cav-3 (green) at CM plasma membrane (large open arrow), FB (nucleus: large asterisks) interconnecting with tails (small open arrow) and CEC (nucleus: small asterisks). Also, Cav-1 immunoreactivity occurs near CM Z-lines and partial co-localization with Cav-3 (E–G). Cav-2 (green) co-localizes partially with Cav-3 (red) at CEC (arrows). CEC make frequently anastomotic structure (asterisk) (I–K). In Cav-1−/− Cav-2 (green) immunore-activity occurs regardless of Cav-1 at CEC (d). Cav-3 (green), immunoreactivity occurs also at CM plasma membranes, Z-lines and FB (asterisks) (H). Co-localization (open arrows) of Cav-2 (green) and Cav-3 (red) at CEC is similar to that in Cav-1+/+(I). CM is cardiomyocyte and IS is interstitial space. Scale bar is 10 mm for all images.

Mentions: As noted above, Cav-1 immunoreactivity appeared at plasma membranes and around Z-lines of CM, at FB and at CEC of Cav-1+/+ mouse LVM. Cav-1 immunoreactivity co-localized with Cav-2 only at CEC. 3D structure of FB with Cav-1 and CEC with Cav-2 showed that these cells twisted around each other and all were closely related (Fig. 3A–C). Cav-2 immunoreactivity appeared at CEC of Cav-1−/− mouse LVM similar to that of Cav-1+/+ mouse LVM (Fig. 3D).


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 between caveolins in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ 3D reconstructed images (-45° rotation of y-axis) shows Cav-1 (red) localizes at both FB (nucleus: large asterisks) and CEC (nucleus: small asterisks), but Cav-2 (green) localizes only at CEC. Partial co-localization of Cav-1 and Cav-2 occurs at CEC (open arrows), but sometimes Cav-1 without Cav-2 occurs at CEC membrane (closed arrows) (A–C). Cav-1 (red) co-localizes well with Cav-3 (green) at CM plasma membrane (large open arrow), FB (nucleus: large asterisks) interconnecting with tails (small open arrow) and CEC (nucleus: small asterisks). Also, Cav-1 immunoreactivity occurs near CM Z-lines and partial co-localization with Cav-3 (E–G). Cav-2 (green) co-localizes partially with Cav-3 (red) at CEC (arrows). CEC make frequently anastomotic structure (asterisk) (I–K). In Cav-1−/− Cav-2 (green) immunore-activity occurs regardless of Cav-1 at CEC (d). Cav-3 (green), immunoreactivity occurs also at CM plasma membranes, Z-lines and FB (asterisks) (H). Co-localization (open arrows) of Cav-2 (green) and Cav-3 (red) at CEC is similar to that in Cav-1+/+(I). CM is cardiomyocyte and IS is interstitial space. Scale bar is 10 mm for all images.
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Related In: Results  -  Collection

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fig03: Co-localization between caveolins in Cav-1+/+ and Cav-1−/− mouse LVM. In Cav-1+/+ 3D reconstructed images (-45° rotation of y-axis) shows Cav-1 (red) localizes at both FB (nucleus: large asterisks) and CEC (nucleus: small asterisks), but Cav-2 (green) localizes only at CEC. Partial co-localization of Cav-1 and Cav-2 occurs at CEC (open arrows), but sometimes Cav-1 without Cav-2 occurs at CEC membrane (closed arrows) (A–C). Cav-1 (red) co-localizes well with Cav-3 (green) at CM plasma membrane (large open arrow), FB (nucleus: large asterisks) interconnecting with tails (small open arrow) and CEC (nucleus: small asterisks). Also, Cav-1 immunoreactivity occurs near CM Z-lines and partial co-localization with Cav-3 (E–G). Cav-2 (green) co-localizes partially with Cav-3 (red) at CEC (arrows). CEC make frequently anastomotic structure (asterisk) (I–K). In Cav-1−/− Cav-2 (green) immunore-activity occurs regardless of Cav-1 at CEC (d). Cav-3 (green), immunoreactivity occurs also at CM plasma membranes, Z-lines and FB (asterisks) (H). Co-localization (open arrows) of Cav-2 (green) and Cav-3 (red) at CEC is similar to that in Cav-1+/+(I). CM is cardiomyocyte and IS is interstitial space. Scale bar is 10 mm for all images.
Mentions: As noted above, Cav-1 immunoreactivity appeared at plasma membranes and around Z-lines of CM, at FB and at CEC of Cav-1+/+ mouse LVM. Cav-1 immunoreactivity co-localized with Cav-2 only at CEC. 3D structure of FB with Cav-1 and CEC with Cav-2 showed that these cells twisted around each other and all were closely related (Fig. 3A–C). Cav-2 immunoreactivity appeared at CEC of Cav-1−/− mouse LVM similar to that of Cav-1+/+ mouse LVM (Fig. 3D).

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