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Macrophage Receptor with Collagenous Structure (MARCO) Is Processed by either Macropinocytosis or Endocytosis-Autophagy Pathway.

Hirano S, Kanno S - PLoS ONE (2015)

Bottom Line: The Macrophage Receptor with COllagenous structure (MARCO) protein is a plasma membrane receptor for un-opsonized or environmental particles on phagocytic cells.The small puncta trafficked between the peri-nuclear region and the distal ends of cells back and forth at rates of up to 2-3 μm/sec; tubulin, but not actin, regulated the trafficking of the small puncta.Besides phagocytosis MARCO, an adhesive plasma membrane receptor, may play a role in incorporation of various extracellular materials into the cell via both macropinocytic and endocytic pathways.

View Article: PubMed Central - PubMed

Affiliation: NanoTox, RCER, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506, Japan.

ABSTRACT
The Macrophage Receptor with COllagenous structure (MARCO) protein is a plasma membrane receptor for un-opsonized or environmental particles on phagocytic cells. Here, we show that MARCO was internalized either by ruffling of plasma membrane followed by macropinocytosis or by endocytosis followed by fusion with autophagosome in CHO-K1 cells stably transfected with GFP-MARCO. The macropinocytic process generated large vesicles when the plasma membrane subsided. The endocytosis/autophagosome (amphisome) generated small fluorescent puncta which were visible in the presence of glutamine, chloroquine, bafilomycin, ammonia, and other amines. The small puncta, but not the large vesicles, co-localized with LC3B and lysosomes. The LC3-II/LC3-I ratio increased in the presence of glutamine, ammonia, and chloroquine in various cells. The small puncta trafficked between the peri-nuclear region and the distal ends of cells back and forth at rates of up to 2-3 μm/sec; tubulin, but not actin, regulated the trafficking of the small puncta. Besides phagocytosis MARCO, an adhesive plasma membrane receptor, may play a role in incorporation of various extracellular materials into the cell via both macropinocytic and endocytic pathways.

No MeSH data available.


Related in: MedlinePlus

Macropinocytic activity of MARCO-GFP CHO cells.(A) The cells were grown in glass-bottom culture dishes in F12 complete medium and observed using confocal microscopy. Fluorescence images were recorded at 1-min intervals. The arrows and arrowheads indicate the formation and fate of two separate macropinocytic sites. See also S1 Movie. (B) Scanning electron micrographs of GFP-MARCO-CHO cells. The right panel provides a higher magnification image of the boxed area indicated in the left panel. The arrow indicates the macropinocytic structure.
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pone.0142062.g001: Macropinocytic activity of MARCO-GFP CHO cells.(A) The cells were grown in glass-bottom culture dishes in F12 complete medium and observed using confocal microscopy. Fluorescence images were recorded at 1-min intervals. The arrows and arrowheads indicate the formation and fate of two separate macropinocytic sites. See also S1 Movie. (B) Scanning electron micrographs of GFP-MARCO-CHO cells. The right panel provides a higher magnification image of the boxed area indicated in the left panel. The arrow indicates the macropinocytic structure.

Mentions: GFP-MARCO was internalized by macropinocytosis in GFP-MARCO-CHO cells, as shown in Fig 1A and 1B and S1 Movie. Dense GFP labeling or local ruffling of plasma membrane appeared in peripheral sites of the cell as plasma membrane ruffled and the GFP-labeled membrane was pinched off and internalized as vesicles. The vesicles disappeared within several minutes.


Macrophage Receptor with Collagenous Structure (MARCO) Is Processed by either Macropinocytosis or Endocytosis-Autophagy Pathway.

Hirano S, Kanno S - PLoS ONE (2015)

Macropinocytic activity of MARCO-GFP CHO cells.(A) The cells were grown in glass-bottom culture dishes in F12 complete medium and observed using confocal microscopy. Fluorescence images were recorded at 1-min intervals. The arrows and arrowheads indicate the formation and fate of two separate macropinocytic sites. See also S1 Movie. (B) Scanning electron micrographs of GFP-MARCO-CHO cells. The right panel provides a higher magnification image of the boxed area indicated in the left panel. The arrow indicates the macropinocytic structure.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4636388&req=5

pone.0142062.g001: Macropinocytic activity of MARCO-GFP CHO cells.(A) The cells were grown in glass-bottom culture dishes in F12 complete medium and observed using confocal microscopy. Fluorescence images were recorded at 1-min intervals. The arrows and arrowheads indicate the formation and fate of two separate macropinocytic sites. See also S1 Movie. (B) Scanning electron micrographs of GFP-MARCO-CHO cells. The right panel provides a higher magnification image of the boxed area indicated in the left panel. The arrow indicates the macropinocytic structure.
Mentions: GFP-MARCO was internalized by macropinocytosis in GFP-MARCO-CHO cells, as shown in Fig 1A and 1B and S1 Movie. Dense GFP labeling or local ruffling of plasma membrane appeared in peripheral sites of the cell as plasma membrane ruffled and the GFP-labeled membrane was pinched off and internalized as vesicles. The vesicles disappeared within several minutes.

Bottom Line: The Macrophage Receptor with COllagenous structure (MARCO) protein is a plasma membrane receptor for un-opsonized or environmental particles on phagocytic cells.The small puncta trafficked between the peri-nuclear region and the distal ends of cells back and forth at rates of up to 2-3 μm/sec; tubulin, but not actin, regulated the trafficking of the small puncta.Besides phagocytosis MARCO, an adhesive plasma membrane receptor, may play a role in incorporation of various extracellular materials into the cell via both macropinocytic and endocytic pathways.

View Article: PubMed Central - PubMed

Affiliation: NanoTox, RCER, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506, Japan.

ABSTRACT
The Macrophage Receptor with COllagenous structure (MARCO) protein is a plasma membrane receptor for un-opsonized or environmental particles on phagocytic cells. Here, we show that MARCO was internalized either by ruffling of plasma membrane followed by macropinocytosis or by endocytosis followed by fusion with autophagosome in CHO-K1 cells stably transfected with GFP-MARCO. The macropinocytic process generated large vesicles when the plasma membrane subsided. The endocytosis/autophagosome (amphisome) generated small fluorescent puncta which were visible in the presence of glutamine, chloroquine, bafilomycin, ammonia, and other amines. The small puncta, but not the large vesicles, co-localized with LC3B and lysosomes. The LC3-II/LC3-I ratio increased in the presence of glutamine, ammonia, and chloroquine in various cells. The small puncta trafficked between the peri-nuclear region and the distal ends of cells back and forth at rates of up to 2-3 μm/sec; tubulin, but not actin, regulated the trafficking of the small puncta. Besides phagocytosis MARCO, an adhesive plasma membrane receptor, may play a role in incorporation of various extracellular materials into the cell via both macropinocytic and endocytic pathways.

No MeSH data available.


Related in: MedlinePlus