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Tubular lysosomes accompany stimulated pinocytosis in macrophages
Abstract:
A network of tubular lysosomes extends through the cytoplasm of J774.2 macrophages and phorbol ester-treated mouse peritoneal macrophages. The presence of this network is dependent upon the integrity of cytoplasmic microtubules and correlates with high cellular rates of accumulation of Lucifer Yellow (LY), a marker of fluid phase pinocytosis. We tested the hypothesis that the efficiency of LY transfer between the pinosomal and lysosomal compartments is increased in the presence of tubular lysosomes by asking how conditions that deplete the tubular lysosome network affect pinocytic accumulation of LY. Tubular lysosomes were disassembled in cells treated with microtubule-depolymerizing drugs or in cells that had phagocytosed latex beads. In unstimulated peritoneal macrophages, which normally contain few tubular lysosomes and which exhibit relatively inefficient transfer of pinocytosed LY to lysosomes, such treatments had little effect on pinocytosis. However, in J774 macrophages and phorbol ester-stimulated peritoneal macrophages, these treatments markedly reduced the efficiency of pinocytic accumulation of LY. We conclude that a basal level of solute accumulation via pinocytosis proceeds independently of the tubular lysosomes, and that an extended tubular lysosomal network contributes to the elevated rates of solute accumulation that accompany macrophage stimulation. Moreover, we suggest that the transformed mouse macrophage cell line J774 exhibits this stimulated pinocytosis constitutively.
Insights
Tubular lysosomes enhance fluid uptake in macrophages. Disrupting this network reduces pinocytosis efficiency in stimulated cells, indicating its crucial role in macrophage solute accumulation.
Area of Science:
- Cell Biology
- Immunology
- Macrophage Biology
Background:
- Lysosomes are key organelles for cellular waste disposal and nutrient recycling.
- Macrophage pinocytosis is a vital process for fluid and solute uptake.
- Tubular lysosomes form an extensive network within certain macrophage types.
Purpose of the Study:
- To investigate the role of tubular lysosomes in fluid phase pinocytosis.
- To determine if tubular lysosomes enhance the efficiency of solute accumulation in macrophages.
- To understand how microtubule integrity affects lysosomal networks and pinocytosis.
Main Methods:
- Utilized J774.2 and phorbol ester-stimulated mouse peritoneal macrophages.
- Disrupted tubular lysosome networks using microtubule-depolymerizing drugs and latex bead phagocytosis.
- Quantified fluid phase pinocytosis by measuring Lucifer Yellow (LY) accumulation.
Main Results:
- Disrupting tubular lysosomes reduced pinocytic accumulation of LY in J774.2 and stimulated peritoneal macrophages.
- Unstimulated peritoneal macrophages, with limited tubular lysosomes, showed minimal impact from network disruption.
- The presence of an extended tubular lysosomal network correlated with high rates of LY accumulation.
Conclusions:
- A basal level of pinocytosis occurs independently of tubular lysosomes.
- An extended tubular lysosomal network significantly contributes to elevated solute accumulation in stimulated macrophages.
- The J774 macrophage cell line constitutively exhibits stimulated pinocytosis.