Limits...
Leishmania donovani infection enhances lateral mobility of macrophage membrane protein which is reversed by liposomal cholesterol.

Ghosh M, Roy K, Das Mukherjee D, Chakrabarti G, Roy Choudhury K, Roy S - PLoS Negl Trop Dis (2014)

Bottom Line: The protozoan parasite Leishmania donovani (LD) reduces cellular cholesterol of the host possibly for its own benefit.To our knowledge this is the first direct demonstration that LD parasites during their intracellular life cycle increases lateral mobility of membrane proteins and decreases F-actin level in infected macrophages.Such defects may contribute to ineffective intracellular signaling and other cellular functions.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases and Immunology Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India.

ABSTRACT

Background: The protozoan parasite Leishmania donovani (LD) reduces cellular cholesterol of the host possibly for its own benefit. Cholesterol is mostly present in the specialized compartment of the plasma membrane. The relation between mobility of membrane proteins and cholesterol depletion from membrane continues to be an important issue. The notion that leishmania infection alters the mobility of membrane proteins stems from our previous study where we showed that the distance between subunits of IFNγ receptor (R1 and R2) on the cell surface of LD infected cell is increased, but is restored to normal by liposomal cholesterol treatment.

Methodology/principal findings: We determined the lateral mobility of a membrane protein in normal, LD infected and liposome treated LD infected cells using GFP-tagged PLCδ1 as a probe. The mobility of PLCδ1 was computationally analyzed from the time lapse experiment using boundary distance plot and radial profile movement. Our results showed that the lateral mobility of the membrane protein, which is increased in infection, is restored to normal upon liposomal cholesterol treatment. The results of FRAP experiment lent further credence to the above notion. The membrane proteins are intimately linked with cellular actin and alteration of cellular actin may influence lateral mobility. We found that F-actin is decreased in infection but is restored to normal upon liposomal cholesterol treatment as evident from phalloidin staining and also from biochemical analysis by immunoblotting.

Conclusions/significances: To our knowledge this is the first direct demonstration that LD parasites during their intracellular life cycle increases lateral mobility of membrane proteins and decreases F-actin level in infected macrophages. Such defects may contribute to ineffective intracellular signaling and other cellular functions.

Show MeSH

Related in: MedlinePlus

Confocal images of plcδ1-gfp transfected RAW 264.7 cells followed by infection with LD (I-MΦ).The expression of PLCδ1-GFP (green) is on the cell surface. The cells were stained with Hoechst 33342 (blue). Parasites are marked by blue dot and indicated by arrow.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4256160&req=5

pntd-0003367-g001: Confocal images of plcδ1-gfp transfected RAW 264.7 cells followed by infection with LD (I-MΦ).The expression of PLCδ1-GFP (green) is on the cell surface. The cells were stained with Hoechst 33342 (blue). Parasites are marked by blue dot and indicated by arrow.

Mentions: For this investigation RAW264.7 cells were used as host cell for infection with LD. The number of intracellular LD and integrity of the nucleus in infected MΦ (I-MΦ) were determined by staining with Hoechst 33342 or DAPI. The intracellular parasite number, verified by Giemsa staining (S1 Figure), was ∼8–9/cell, similar to the results obtained by Hoechst 33342 or DAPI stain (Fig. 1). It was clear that the morphology of the cells and the integrity of nucleus did not change upon infection.


Leishmania donovani infection enhances lateral mobility of macrophage membrane protein which is reversed by liposomal cholesterol.

Ghosh M, Roy K, Das Mukherjee D, Chakrabarti G, Roy Choudhury K, Roy S - PLoS Negl Trop Dis (2014)

Confocal images of plcδ1-gfp transfected RAW 264.7 cells followed by infection with LD (I-MΦ).The expression of PLCδ1-GFP (green) is on the cell surface. The cells were stained with Hoechst 33342 (blue). Parasites are marked by blue dot and indicated by arrow.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256160&req=5

pntd-0003367-g001: Confocal images of plcδ1-gfp transfected RAW 264.7 cells followed by infection with LD (I-MΦ).The expression of PLCδ1-GFP (green) is on the cell surface. The cells were stained with Hoechst 33342 (blue). Parasites are marked by blue dot and indicated by arrow.
Mentions: For this investigation RAW264.7 cells were used as host cell for infection with LD. The number of intracellular LD and integrity of the nucleus in infected MΦ (I-MΦ) were determined by staining with Hoechst 33342 or DAPI. The intracellular parasite number, verified by Giemsa staining (S1 Figure), was ∼8–9/cell, similar to the results obtained by Hoechst 33342 or DAPI stain (Fig. 1). It was clear that the morphology of the cells and the integrity of nucleus did not change upon infection.

Bottom Line: The protozoan parasite Leishmania donovani (LD) reduces cellular cholesterol of the host possibly for its own benefit.To our knowledge this is the first direct demonstration that LD parasites during their intracellular life cycle increases lateral mobility of membrane proteins and decreases F-actin level in infected macrophages.Such defects may contribute to ineffective intracellular signaling and other cellular functions.

View Article: PubMed Central - PubMed

Affiliation: Infectious Diseases and Immunology Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India.

ABSTRACT

Background: The protozoan parasite Leishmania donovani (LD) reduces cellular cholesterol of the host possibly for its own benefit. Cholesterol is mostly present in the specialized compartment of the plasma membrane. The relation between mobility of membrane proteins and cholesterol depletion from membrane continues to be an important issue. The notion that leishmania infection alters the mobility of membrane proteins stems from our previous study where we showed that the distance between subunits of IFNγ receptor (R1 and R2) on the cell surface of LD infected cell is increased, but is restored to normal by liposomal cholesterol treatment.

Methodology/principal findings: We determined the lateral mobility of a membrane protein in normal, LD infected and liposome treated LD infected cells using GFP-tagged PLCδ1 as a probe. The mobility of PLCδ1 was computationally analyzed from the time lapse experiment using boundary distance plot and radial profile movement. Our results showed that the lateral mobility of the membrane protein, which is increased in infection, is restored to normal upon liposomal cholesterol treatment. The results of FRAP experiment lent further credence to the above notion. The membrane proteins are intimately linked with cellular actin and alteration of cellular actin may influence lateral mobility. We found that F-actin is decreased in infection but is restored to normal upon liposomal cholesterol treatment as evident from phalloidin staining and also from biochemical analysis by immunoblotting.

Conclusions/significances: To our knowledge this is the first direct demonstration that LD parasites during their intracellular life cycle increases lateral mobility of membrane proteins and decreases F-actin level in infected macrophages. Such defects may contribute to ineffective intracellular signaling and other cellular functions.

Show MeSH
Related in: MedlinePlus