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MCP-1 downregulates MMP-9 export via vesicular redistribution to lysosomes in rat portal fibroblasts.

Hickman DA, Syal G, Fausther M, Lavoie EG, Goree JR, Storrie B, Dranoff JA - Physiol Rep (2014)

Bottom Line: We examined the effect of MCP-1 on release of matrix metalloproteinase-9 (MMP-9) by rat PF.We found that MCP-1 blocks PF release of MMP-9 in a posttranslational fashion.Our data demonstrated that, in the presence of MCP-1, MMP-9-containing vesicles were shunted to a lysosome-like compartment.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.

No MeSH data available.


Related in: MedlinePlus

Effect of MCP‐1 on MMP‐9/LAMP‐1 colocalization in PF. PF were untreated or treated with MCP‐1 (10 ng/mL) ± blocking antibody (bAb) overnight then fixed. MMP‐9 and LAMP‐1 were detected by immunofluorescence, and nuclei were labeled with TOPRO. MMP‐9 fluorescence is pseudocolored red, LAMP‐1 fluorescence is pseudocolored green, and TOPRO fluorescence is pseudocolored green. In control cells or cells treated with MCP‐1 + bAb, no MMP‐9 intracellular fluorescence is noted. LAMP‐1 fluorescence is seen throughout the cell, with a concentration in the perinuclear cytoplasm. In contrast, PF treated with MCP‐1 demonstrate a large amount of MMP‐9 in a vesicular distribution, which overlies the LAMP‐1 fluorescence.
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fig06: Effect of MCP‐1 on MMP‐9/LAMP‐1 colocalization in PF. PF were untreated or treated with MCP‐1 (10 ng/mL) ± blocking antibody (bAb) overnight then fixed. MMP‐9 and LAMP‐1 were detected by immunofluorescence, and nuclei were labeled with TOPRO. MMP‐9 fluorescence is pseudocolored red, LAMP‐1 fluorescence is pseudocolored green, and TOPRO fluorescence is pseudocolored green. In control cells or cells treated with MCP‐1 + bAb, no MMP‐9 intracellular fluorescence is noted. LAMP‐1 fluorescence is seen throughout the cell, with a concentration in the perinuclear cytoplasm. In contrast, PF treated with MCP‐1 demonstrate a large amount of MMP‐9 in a vesicular distribution, which overlies the LAMP‐1 fluorescence.

Mentions: On the basis of the above observations, we attempted to determine directly whether MCP‐1 induced trafficking of endogenously expressed MMP‐9 to lysosomes. We did this using two separate approaches. First, we observed the distribution of MMP‐9 at the subcellular level in the presence or absence of MCP‐1 using immuno‐EM (Fig. 5). In control cells, MMP‐9 can be seen in vesicles near the plasma membrane; however, in PF treated with MCP‐1, MMP‐9 is seen in structures with typical features of lysosomes. Second, we determined whether MCP‐1 induced trafficking of MMP‐9 vesicles to regions expressing the lysosomal glyocoprotein LAMP‐1 (Eskelinen 2006) (Fig. 6). As noted previously in Figure 3, control cells or cells treated with MCP‐1 + bAb did not show MMP‐9 intracellular fluorescence. However, PF treated with MCP‐1 showed clusters of MMP‐9 in the perinuclear cytoplasm that colocalized with LAMP‐1. Taken together, these studies provided evidence that MCP‐1 shifts trafficking of MMP‐9 to lysosomes.


MCP-1 downregulates MMP-9 export via vesicular redistribution to lysosomes in rat portal fibroblasts.

Hickman DA, Syal G, Fausther M, Lavoie EG, Goree JR, Storrie B, Dranoff JA - Physiol Rep (2014)

Effect of MCP‐1 on MMP‐9/LAMP‐1 colocalization in PF. PF were untreated or treated with MCP‐1 (10 ng/mL) ± blocking antibody (bAb) overnight then fixed. MMP‐9 and LAMP‐1 were detected by immunofluorescence, and nuclei were labeled with TOPRO. MMP‐9 fluorescence is pseudocolored red, LAMP‐1 fluorescence is pseudocolored green, and TOPRO fluorescence is pseudocolored green. In control cells or cells treated with MCP‐1 + bAb, no MMP‐9 intracellular fluorescence is noted. LAMP‐1 fluorescence is seen throughout the cell, with a concentration in the perinuclear cytoplasm. In contrast, PF treated with MCP‐1 demonstrate a large amount of MMP‐9 in a vesicular distribution, which overlies the LAMP‐1 fluorescence.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4255798&req=5

fig06: Effect of MCP‐1 on MMP‐9/LAMP‐1 colocalization in PF. PF were untreated or treated with MCP‐1 (10 ng/mL) ± blocking antibody (bAb) overnight then fixed. MMP‐9 and LAMP‐1 were detected by immunofluorescence, and nuclei were labeled with TOPRO. MMP‐9 fluorescence is pseudocolored red, LAMP‐1 fluorescence is pseudocolored green, and TOPRO fluorescence is pseudocolored green. In control cells or cells treated with MCP‐1 + bAb, no MMP‐9 intracellular fluorescence is noted. LAMP‐1 fluorescence is seen throughout the cell, with a concentration in the perinuclear cytoplasm. In contrast, PF treated with MCP‐1 demonstrate a large amount of MMP‐9 in a vesicular distribution, which overlies the LAMP‐1 fluorescence.
Mentions: On the basis of the above observations, we attempted to determine directly whether MCP‐1 induced trafficking of endogenously expressed MMP‐9 to lysosomes. We did this using two separate approaches. First, we observed the distribution of MMP‐9 at the subcellular level in the presence or absence of MCP‐1 using immuno‐EM (Fig. 5). In control cells, MMP‐9 can be seen in vesicles near the plasma membrane; however, in PF treated with MCP‐1, MMP‐9 is seen in structures with typical features of lysosomes. Second, we determined whether MCP‐1 induced trafficking of MMP‐9 vesicles to regions expressing the lysosomal glyocoprotein LAMP‐1 (Eskelinen 2006) (Fig. 6). As noted previously in Figure 3, control cells or cells treated with MCP‐1 + bAb did not show MMP‐9 intracellular fluorescence. However, PF treated with MCP‐1 showed clusters of MMP‐9 in the perinuclear cytoplasm that colocalized with LAMP‐1. Taken together, these studies provided evidence that MCP‐1 shifts trafficking of MMP‐9 to lysosomes.

Bottom Line: We examined the effect of MCP-1 on release of matrix metalloproteinase-9 (MMP-9) by rat PF.We found that MCP-1 blocks PF release of MMP-9 in a posttranslational fashion.Our data demonstrated that, in the presence of MCP-1, MMP-9-containing vesicles were shunted to a lysosome-like compartment.

View Article: PubMed Central - PubMed

Affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock, Arkansas.

No MeSH data available.


Related in: MedlinePlus