IFN-γ-induced increase in the mobility of MHC class II compartments in astrocytes depends on intermediate filaments
© Vardjan et al.; licensee BioMed Central Ltd. 2012
Received: 7 April 2012
Accepted: 28 May 2012
Published: 26 June 2012
In immune-mediated diseases of the central nervous system, astrocytes exposed to interferon-γ (IFN-γ) can express major histocompatibility complex (MHC) class II molecules and antigens on their surface. MHC class II molecules are thought to be delivered to the cell surface by membrane-bound vesicles. However, the characteristics and dynamics of this vesicular traffic are unclear, particularly in reactive astrocytes, which overexpress intermediate filament (IF) proteins that may affect trafficking. The aim of this study was to determine the mobility of MHC class II vesicles in wild-type (WT) astrocytes and in astrocytes devoid of IFs.
The identity of MHC class II compartments in WT and IF-deficient astrocytes 48 h after IFN-γ activation was determined immunocytochemically by using confocal microscopy. Time-lapse confocal imaging and Alexa Fluor546-dextran labeling of late endosomes/lysosomes in IFN-γ treated cells was used to characterize the motion of MHC class II vesicles. The mobility of vesicles was analyzed using ParticleTR software.
Confocal imaging of primary cultures of WT and IF-deficient astrocytes revealed IFN-γ induced MHC class II expression in late endosomes/lysosomes, which were specifically labeled with Alexa Fluor546-conjugated dextran. Live imaging revealed faster movement of dextran-positive vesicles in IFN-γ-treated than in untreated astrocytes. Vesicle mobility was lower in IFN-γ-treated IF-deficient astrocytes than in WT astrocytes. Thus, the IFN-γ-induced increase in the mobility of MHC class II compartments is IF-dependent.
Since reactivity of astrocytes is a hallmark of many CNS pathologies, it is likely that the up-regulation of IFs under such conditions allows a faster and therefore a more efficient delivery of MHC class II molecules to the cell surface. In vivo, such regulatory mechanisms may enable antigen-presenting reactive astrocytes to respond rapidly and in a controlled manner to CNS inflammation.
KeywordsAstrocytes Vesicle mobility Late endosomes/lysosomes Major histocompatibility class II compartments Interferon-γ Dextran labeling Immune response
Antigen-presenting cells (APCs) are key players in the immune response. They take up and process exogenous antigens and present them to CD4 helper T-cells, resulting in antigen-specific T-cell activation. After endocytosis of antigens by APCs, early endosomes gradually transform into late endosomes/lysosomes, where antigens are processed to peptides and loaded onto MHC class II molecules. Late endosomes/lysosomes expressing peptide-MHC class II complexes are delivered to the cell surface for recognition by T-cell receptors on CD4 helper T-cells [1, 2].
Astrocytes, the most abundant glial cells in the central nervous system (CNS), can act as ‘non-professional’ APCs. Unlike professional APCs (for example, dendritic cells, macrophages, B-cells), astrocytes express cell-surface MHC class II molecules only upon exposure to the cytokine IFN-γ [3, 4]. IFN-γ-activated astrocytes participate in antigen presentation and activation of CD4 helper T-cells in immune-mediated CNS disorders such as multiple sclerosis [5, 6] and in experimental autoimmune encephalomyelitis . Expression of MHC II molecules in astrocytes is thought to be controlled by neurons, since IFN-γ induces MHC class II expression only in slice culture zones containing degenerated neurons, in which spontaneous neuronal activity (for example, release of glutamate or noradrenaline) is suppressed [7, 8]. In primary astrocyte cultures without neurons, IFN-γ induces expression of surface MHC class II and some APC-specific co-stimulatory molecules [9, 10]. Such cultures may be useful for studying antigen presentation at the subcellular level.
The trafficking of MHC class II compartments in APCs is dependent on the cytoskeletal network. This trafficking is influenced by actin microfilaments  and actin-based motor proteins in B-cells  and by microtubules and microtubule-based motor proteins in dendritic cells and a human melanoma cell line [13, 14]. The MHC class II compartments in astrocytes and their mobility have not been characterized.
In reactive astrocytes, which are found in almost all CNS pathologies, the expression of IF proteins (cell-type-specific components of the cytoskeleton)  is up-regulated [16, 17]. Studies of primary astrocytes from mice deficient in the IF proteins glial fibrillary acidic protein and vimentin (GFAP -/- Vim -/- ) and completely devoid of astrocyte cytosolic IFs [18, 19] have suggested that the up-regulation of IFs in pathological situations may deregulate vesicle trafficking [19, 20].
In this study, we sought to determine whether the trafficking of MHC class II compartments, which is closely associated with neuroinflammation, involves IFs. Live imaging revealed faster movement of dextran-positive vesicles in IFN-γ-treated than in untreated astrocytes. These compartments enriched with MHC class II molecules upon IFN-γ treatment, were less mobile in cytosolic IF-deficient (GFAP -/- Vim -/- ) than in WT astrocytes, indicating that the IFN-γ-induced increase in the mobility of MHC class II compartments in astrocytes depends on IFs.
Astrocyte cultures were prepared from the cerebrum of 1-day-old GFAP -/- Vim -/- and WT mice on a mixed C57Bl6/129Sv/129Ola genetic background and maintained as described [18, 21–23]. GFAP -/- Vim -/- mice [18, 24] were obtained by cross-breeding mice lacking GFAP  and mice lacking Vim . Before experiments, the cells were removed from the culture flasks with trypsin/EDTA and plated on 22-mm glass coverslips coated with poly-L-lysine. Cells were maintained in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 1 mM sodium pyruvate, 2 mM L-glutamine, and 25 μg/mL penicillin-streptomycin in an atmosphere of humidified air (95%) and CO2 (5%). In some experiments, astrocytes were treated with 600 U/ml IFN-γ (0.06 μg/mL; Abcam, Cambridge, UK) for 48 h at 37°C. The purity of astrocyte cultures was confirmed with antibodies against astrocytic markers GFAP (Sigma Aldrich, St Louis, MO, USA) or glutamine synthetase (Abcam, Cambridge, MA, USA). All chemicals were from Sigma Aldrich (St Louis, MO, USA) unless noted otherwise.
The care for experimental animals was in accordance with International Guiding Principles for Biomedical Research Involving Animals developed by the Council for International Organizations of Medical Sciences and Directive on Conditions for issue of License for Animal Experiments for Scientific Research Purposes (Official Gazette of the RS, No. 43/07).
Cell-surface expression of MHC class II molecules was determined by flow cytometry (FACSCalibur, BD Biosciences, Franklin Lakes, NJ, USA) and fluorescence-labeled rat Alexa Fluor488 anti-mouse I-A/I-E (MHC class II) antibodies (BioLegend, San Diego, CA, USA). Control and IFN-γ-activated mouse WT and GFAP -/- Vim -/- astrocytes were removed from flasks with trypsin/EDTA and collected by centrifugation. Antibody was added, and the cells were incubated at room temperature for 15 min in the dark, washed twice with Dulbecco’s phosphate-buffered saline, and resuspended in 2% paraformaldehyde. Alexa Fluor488-IgG2b cocktail (BioLegend) was used as an isotype control. From a forward scatter and side scatter dot plot, the population of cells was gated for further analysis. Cell-surface expression of MHC class II molecules was determined by fluorescence-activated cell sorting.
Control and IFN-γ-activated astrocytes growing on coverslips were fixed in 2% paraformaldehyde for 5 min at room temperature and treated with 10% goat serum for 1 h at 37°C. In some experiments, astrocytes were incubated with fixable 10-kDa Alexa Fluor546 dextran conjugate (0.1 mg/mL; Molecular Probes, Invitrogen, Eugene, OR, USA) for 16 h at 37°C and washed for 3 h at 37°C. Cultures were then stained with one or both of the following primary antibodies for 2 h at 37°C: Alexa Fluor488 rat anti-mouse MHC class II (1:200) and rabbit anti-lysosomal-associated membrane protein 1 (LAMP1; 1:250; Abcam). Excess primary antibody was washed off, and the cultures were incubated for 1 h at 37°C with Alexa Fluor546- or Alexa Fluor488-conjugated secondary goat anti-rabbit IgG antibody (1:600; Abcam). Excess antibody was removed, and cells were treated with SlowFade Gold antifade reagent (Molecular Probes). Immunolabeled cells were imaged with an inverted Zeiss LSM 510 or Zeiss LSM 510 Meta confocal microscopes with an oil-immersion plan apochromatic objective (63x, 1.4 NA; Carl Zeiss, Jena, Germany), using 488-nm Ar-Ion and 543-nm He-Ne laser excitation. Emission light was acquired sequentially with a 505-530-nm bandpass emission filter (Alexa Fluor488) and a 560-nm long-pass emission filter (Alexa Fluor546).
Time-lapse confocal imaging
Astrocytes were loaded with 0.1 mg/mL Alexa Fluor546-dextran (Molecular Probes) for 16 h at 37°C, washed for 3 h at 37°C, and transferred to the chamber for imaging. Time-lapse fluorescence images (512 x 512 pixels) were obtained every 2 s for 3 min (90 frames) with the Zeiss LSM 510 Meta confocal microscope described above and 543-nm He-Ne laser excitation. Emission light was acquired with a 560-nm long-pass emission filter. Sixty seconds after the start of recording cells were superfused with 1 mM ATP in a standard saline solution (10 mM Hepes/NaOH, pH 7.2, 10 mM D-glucose, 131.8 mM NaCl, 1.8 mM CaCl2, 2 mM MgCl2, and 5 mM KCl).
The motion of Alexa Fluor546-dextran–labeled vesicles before and 30 s after the start of ATP stimulation was tracked in exported TIFF files by ParticleTR software (Celica, Ljubljana, Slovenia) as described . A single dextran-labeled vesicle was selected in the image sequence. Using a simplex algorithm with a least-squares estimator, the software fits a 2D Gaussian curve to the vesicle intensity profile. The peak of curve was recorded as the x,y coordinates of the vesicle position. The time (from the beginning of tracking for a single vesicle), step length (displacement of a vesicle over 2 s), TL (total length of the analyzed vesicle pathway), and MD (measure of a net translocation of vesicle) were determined [19, 23]. Vesicle mobility in control and IFN-γ-activated astrocytes before and after ATP stimulation was analyzed in 40 cells from five independent mouse WT astrocyte cultures and in 33 cells from four independent mouse GFAP -/- Vim -/- astrocyte cultures. Vesicle mobility was analyzed for 30 s (15 frames).
Data are presented as mean ± s.e.m. Differences in TL and MD were analyzed by t test. Differences in the fraction of vesicles exhibiting directional mobility were analyzed with Fisher’s exact test. Differences between the slopes were analyzed by Analysis of covariance. P <0.05 was considered significant.
IFN-γ induces expression of MHC class II molecules on GFAP -/- Vim -/- astrocytes
Fluorescent dextran labels MHC class II–positive late endosomes/lysosomes in WT and GFAP -/- Vim -/- astrocytes
To verify that the dextran-labeled structures were late endosomes/lysosomes, we fixed and stained the cells with antibodies against LAMP1, a marker of late endosomes/lysosomes. In both WT and GFAP -/- Vim -/- astrocytes, internalized Alexa Fluor546-dextran was highly colocalized with LAMP1 (Figure 2). Thus, the lack of IFs in GFAP -/- Vim -/- astrocytes does not affect long-term dextran loading into late endosomes/lysosomes.
Increased mobility of dextran-labeled vesicles in IFN-γ-activated astrocytes requires IFs
Next we used dextran labeling and time-lapse confocal imaging to study the mobility of late endosomes/lysosomes in living astrocytes. To determine whether vesicle mobility in IFN-γ-treated cells is dependent on cytosolic Ca2+ levels, we exposed the cells to 1 mM ATP to increase cytosolic [Ca2+[33, 34]. Cells were imaged every 2 s for 30 s (15 frames) to determine the maximal displacement (MD) and the track length (TL) of vesicles.
Mobility of vesicles in control and IFN-γ-treated WT and GFAP -/- Vim -/- astrocytes under spontaneous conditions and after ATP stimulation
1.73 ± 0.02
0.49 ± 0.01
2.36 ± 0.04a
0.83 ± 0.02a
1.55 ± 0.02b
0.41 ± 0.01b
1.70 ± 0.02b
0.47 ± 0.01b
GFAP -/- Vim -/-
1.51 ± 0.01
0.39 ± 0.01
1.59 ± 0.02a
0.43 ± 0.01a
1.49 ± 0.02
0.37 ± 0.01
1.42 ± 0.02b
0.36 ± 0.01b
After ATP stimulation (Figure 5, Table 1, see also the movies Additional files 5, 6), the mobility of dextran-labeled vesicles was reduced in control and IFN-γ-treated WT astrocytes. In control WT cells, TL decreased by 10.4% and MD by 16.3% (P <0.05). In IFN-γ-treated WT astrocytes, the decreases were larger. TL decreased by 28.0% and MD by 43.4% (P <0.05). Thus, the mobility of dextran-labeled vesicles in control and in IFN-γ-activated WT astrocytes depends on cytosolic [Ca2+].
Stimulation with ATP did not reduce the mobility of dextran-labeled vesicles in control GFAP -/- Vim -/- astrocytes but did in IFN-γ-activated GFAP -/- Vim -/- astrocytes (Figure 5, Table 1; see also the movies Additional files 7, 8). TL was reduced by 10.7% and MD by 16.3% (P <0.05).
IFN-γ increases the proportion of directional vesicles in WT astrocytes
Percentage of directional and non-directional vesicles before and after ATP treatment for control and IFN-γ-treated WT and GFAP -/- Vim -/- astrocytes and slopes of the lines fitted to the vesicle mobility data
Slope (a ± s.e.m.)a
Slope (a ± s.e.m.)a
0.633 ± 0.038c (152)b
0.575 ± 0.020c (426)
0.262 ± 0.006 (1748)
0.219 ± 0.005 (1674)
0.483 ± 0.056c (49)
0.597 ± 0.059c (62)
0.266 ± 0.009 (901)
0.231 ± 0.007 (988)
GFAP -/- Vim -/-
0.581 ± 0.068c (75)
0.553 ± 0.030c (92)
0.324 ± 0.008 (1625)
0.329 ± 0.009 (1508)
0.616 ± 0.069c (33)
0.781 ± 0.148c (20)
0.298 ± 0.010 (817)
0.279 ± 0.012 (780)
The fraction of vesicles with directional mobility was smaller in control and IFN-γ-treated GFAP -/- Vim -/- astrocytes than in control and IFN-γ-treated WT astrocytes (4.4% vs. 8.0% and 5.7% vs. 20.3%, respectively; P <0.001) (Figure 6, upper panels), indicating that IFs affect the directional mobility of vesicles in control [19, 23] and IFN-γ-treated astrocytes.
Stimulation with ATP decreased the percentage of directionally mobile dextran-labeled vesicles in control (8.0% vs. 5.2%, P <0.01) and IFN-γ-treated WT astrocytes (20.3% vs. 5.9%, P <0.001) (Figure 6a), as well as in control (4.4% vs. 3.9%, P = 0.602) and IFN-γ-treated GFAP -/- Vim -/- astrocytes (5.7% vs. 2.5%, P <0.001) (Figure 6b).
This study shows that IFN-γ induces an IF-dependent increase in the mobility of MHC class II compartments in astrocytes. Our findings suggest that up-regulation of IFs in reactive astrocytes allows faster and therefore more efficient delivery of MHC class II molecules to the cell surface.
The expression and targeting of MHC class II molecules in IFN-γ-activated astrocytes
In WT and GFAP -/- Vim -/- primary mouse astrocytes, IFN-γ induced expression of MHC class II molecules on the cell surface and in vesicular structures. As in professional APCs , MHC class II expression colocalized with dextran and LAMP1, which are markers of late endosomes/lysosomes. Thus, although IFs are the key structural and functional element in reactive astrocytes [16, 24, 35] - and may participate in protein targeting defects in polarized epithelial cells , lymphocytes and endothelial cells , dorsal root ganglion neurons , and fibroblasts  - the lack of IFs in astrocytes does not affect the distribution of late endosomal/lysosomal markers or MHC class II expression in these organelles 48 h after IFN-γ activation. Interestingly, the distribution of the late endosomal/lysosomal markers LAMP1 and CD63 is altered in Vim -/- fibroblasts .
IFN-γ induces an IF-dependent increase in the mobility of MHC class II compartments in astrocytes
IFs can deregulate trafficking of vesicular glutamate transporter 1, atrial natriuretic peptide, and acidic vesicles in astrocytes [19, 20] and participate in the transport and distribution of mitochondria in neurons , Golgi complexes in COS-7 cells , late endosomes/lysosomes in fibroblasts , and melanosomes in melanophores . To study the effect of IFs on the trafficking of MHC class II molecules in IFN-γ-activated astrocytes, we used confocal live imaging and dextran loading of late endosomes/lysosomes [28, 29], which are in IFN-γ-activated astrocytes enriched with MHC class II molecules.
Dextran-labeled vesicles moved slower in GFAP -/- Vim -/- astrocytes than in WT astrocytes, indicating that the lack of IFs does not affect the distribution of late endosomes/lysosomes but reduces their mobility. In WT astrocytes, IFN-γ greatly enhanced the mobility of dextran-labeled vesicles, increasing TL by 36.4% and MD by 69.4%. But in GFAP -/- Vim -/- astrocytes, the IFN-γ-induced increase was remarkably attenuated. TL increased by only 5.3% and MD by 10.2%. In WT astrocytes, IFN-γ increased the percentage of directionally mobile vesicles, and some vesicles in both treated and control cells exhibited bidirectional mobility . In IFN-γ-activated GFAP -/- Vim -/- astrocytes, however, the percentage of directionally mobile vesicles increased only slightly. Clearly, IFs help regulate the mobility of MHC class II-positive compartments in IFN-γ-activated astrocytes.
In activated dendritic cells, microtubules drive the directional transport of MHC class II-positive tubules from late endosomes/lysosomes to the plasma membrane [43, 44] and their bidirectional movement . Microtubules and actin filaments also help regulate the mobility of MHC class II compartments in antigen-presenting B-cells [11, 12] and in a human melanoma cell line (Mel JuSo) . Since IFs in various cell types interact with microfilaments, microtubules, and the motor proteins myosin V, kinesin, and dynein [45, 46], the increase in the mobility of MHC class II compartments in IFN-γ-treated astrocytes may reflect up-regulation of IFs [16, 47] and the resulting qualitative or quantitative changes in the interaction between IFs and other components of the cytoskeleton. These three cytoskeletal systems may jointly regulate the trafficking and final positioning of MHC class II molecules in IFN-γ-activated astrocytes.
Proposed (patho)physiological role of IFs in the sorting of MHC class II molecules in IFN-γ-activated astrocytes
Our findings suggest that IFs are involved in the trafficking of MHC class II compartments. Although the lack of IFs attenuated the IFN-γ-induced increase in mobility, MHC class II molecules reached the surface of WT and GFAP -/- Vim -/- astrocytes. What is the role of IFs in the sorting of MHC class II molecules in IFN-γ-activated astrocytes? We propose that up-regulation of IF protein expression [16, 47] in antigen-presenting astrocytes facilitates the transport of antigen-loaded MHC class II molecules from late endosomes/lysosomes in the perinuclear region to the plasma membrane  and the recognition of peptide-MHC class II complexes by T-cell receptors on CD4 helper T-cells.
The pathway by which MHC class II molecules reach the surface of APCs is poorly understood . MHC class II-containing compartments may directly fuse with the plasma membrane [43, 48]. In astrocytes treated with ATP or glutamate, the fluorescence intensity of lysosomes labeled with FM dye decreases in Ca2+-dependent fashion, indicating Ca2+-dependent lysosomal exocytosis in these cells . The authors propose that ATP or glutamate stimulation causes the fusion pore to open briefly (kiss-and-run fusion), releasing ATP molecules stored in lysosomes. In astrocytes, stronger and sustained elevation of [Ca2+i induced with Ca2+ ionophores or by mechanical stimulation triggers predominantly full fusion of lysosomes with the plasma membrane [31, 49]. Both modes of lysosomal exocytosis could deliver lysosomal membrane-bound MHC class II molecules to the plasma membrane [50, 51].
Although ATP did not induce enhanced release of labeled dextran from late endosomes/lysosomes, which would indicate lysosomal fusion [28, 30, 31], it is possible that astrocytes use the lysosomal exocytotic machinery  to deliver MHC class II molecules to the plasma membrane. In lysosomes, the increase in [Ca2+i in response to ATP stimulation is delayed, usually by several minutes [30, 49]. Since we tracked the mobility of dextran-labeled vesicles for 30 s after application of ATP, we could not detect additional lysosomal fusion events. However, when we treated astrocytes with 1 μM adrenalin we did observe the release of labeled dextran from late endosomes/lysosomes (unpublished data), implying a role of cAMP in lysosomal exocytosis. ATP stimulation reduced the mobility of late endosomes/lysosomes. In non-secretory cells, the fraction of mobile lysosomes is reduced after the addition of a Ca2+ ionophore, and in astrocytes, the fraction of Lysotracker-labeled vesicles containing atrial natriuretic peptide is reduced after stimulation with ATP and Ca2+ ionophore [20, 53].
The ATP-induced attenuation of vesicle mobility was more apparent in the presence of IFs (Figure 5), implying a role for IFs in this process . Reduced lysosomal mobility may increase the probability that lysosomes will dock and fuse with the plasma membrane. Thus, in astrocytes functioning as APCs, an increase in [Ca2+i and cAMP (unpublished data) may trigger the delivery of antigen-loaded MHC class II molecules from the lysosomal membrane to the plasma membrane.
The immune privilege of the CNS makes it crucial that the activation of the immune system by resident immunocompetent astrocytes is strictly regulated. We show here that IFN-γ induces the expression of MHC class II molecules in otherwise immunologically silent astrocytes and increases the mobility of vesicles enriched with MHC class II molecules. The increase is highly dependent on IFs, suggesting that the up-regulation of IFs in reactive astrocytes allows faster and therefore more efficient delivery of MHC class II-positive compartments to the cell surface and the activation of CD4 helper T-cells. Our data also suggest that besides IFN-γ activation, astrocytes have a regulatory mechanism, involving increases in [Ca2+]i, and cAMP that controls the fusion of MHC class II compartments with the plasma membrane and the final delivery of MHC class II molecules to the cell surface. In vivo, such regulatory mechanisms may enable antigen-presenting astrocytes to respond rapidly and in a controlled manner to CNS inflammation. For better understanding of the role of astrocytes in CNS pathologies, it will be essential to reveal in greater detail the immunological function of astrocytes and the complex regulatory mechanisms that enable these cells to mediate immune responses in the CNS.
Central nervous system
Glial fibrillary acidic protein
Lysosome-associated membrane protein 1
Major histocompatibility complex
This work was supported by grants P3 310, J34051, J3 4146, J3-0133, L1 2402, and J3-0031 from the Slovenian Research Agency, by the Swedish Medical Research Council (11548), ALF Göteborg (11392), AFA Research Foundation, Sten A. Olsson Foundation for Research and Culture, NanoNet COST Action (to RZ and MP), the EU FP 7 Programs EduGlia (237956 to RZ and MP) and TargetBraIn (279017 to MP).
- Berger AC, Roche PA: MHC class II transport at a glance. J Cell Sci. 2009, 122: 1-4. 10.1242/jcs.035089.PubMed CentralView ArticlePubMed
- Cresswell P: Antigen processing and presentation. Immunol Rev. 2005, 207: 5-7. 10.1111/j.0105-2896.2005.00320.x.View ArticlePubMed
- Shrikant P, Benveniste EN: The central nervous system as an immunocompetent organ: role of glial cells in antigen presentation. J Immunol. 1996, 157: 1819-1822.PubMed
- Hirsch MR, Wietzerbin J, Pierres M, Goridis C: Expression of Ia antigens by cultured astrocytes treated with gamma-interferon. Neurosci Lett. 1983, 41: 199-204. 10.1016/0304-3940(83)90247-1.View ArticlePubMed
- Fontana A, Fierz W, Wekerle H: Astrocytes present myelin basic protein to encephalitogenic T-cell lines. Nature. 1984, 307: 273-276. 10.1038/307273a0.View ArticlePubMed
- Soos JM, Morrow J, Ashley TA, Szente BE, Bikoff EK, Zamvil SS: Astrocytes express elements of the class II endocytic pathway and process central nervous system autoantigen for presentation to encephalitogenic T cells. J Immunol. 1998, 161: 5959-5966.PubMed
- Lee SC, Collins M, Vanguri P, Shin ML: Glutamate differentially inhibits the expression of class II MHC antigens on astrocytes and microglia. J Immunol. 1992, 148: 3391-3397.PubMed
- Neumann H: Control of glial immune function by neurons. Glia. 2001, 36: 191-199. 10.1002/glia.1108.View ArticlePubMed
- Nikcevich KM, Gordon KB, Tan L, Hurst SD, Kroepfl JF, Gardinier M, Barrett TA, Miller SD: IFN-gamma-activated primary murine astrocytes express B7 costimulatory molecules and prime naive antigen-specific T cells. J Immunol. 1997, 158: 614-621.PubMed
- Zeinstra E, Wilczak N, Chesik D, Glazenburg L, Kroese FG, De Keyser J: Simvastatin inhibits interferon-gamma-induced MHC class II up-regulation in cultured astrocytes. J Neuroinflammation. 2006, 3: 16-10.1186/1742-2094-3-16.PubMed CentralView ArticlePubMed
- Barois N, Forquet F, Davoust J: Actin microfilaments control the MHC class II antigen presentation pathway in B cells. J Cell Sci. 1998, 111: 1791-1800.PubMed
- Vascotto F, Lankar D, Faure-André G, Vargas P, Diaz J, Le Roux D, Yuseff MI, Sibarita JB, Boes M, Raposo G, Mougneau E, Glaichenhaus N, Bonnerot C, Manoury B, Lennon-Dumenil AM: The actin-based motor protein myosin II regulates MHC class II trafficking and BCR-driven antigen presentation. J Cell Biol. 2007, 176: 1007-1019. 10.1083/jcb.200611147.PubMed CentralView ArticlePubMed
- Wubbolts R, Fernandez-Borja M, Jordens I, Reits E, Dusseljee S, Echeverri C, Vallee RB, Neefjes J: Opposing motor activities of dynein and kinesin determine retention and transport of MHC class II-containing compartments. J Cell Sci. 1999, 112: 785-795.PubMed
- Vyas JM, Kim YM, Artavanis-Tsakonas K, Love JC, Van der Veen AG, Ploegh HL: Tubulation of class II MHC compartments is microtubule dependent and involves multiple endolysosomal membrane proteins in primary dendritic cells. J Immunol. 2007, 178: 7199-7210.PubMed CentralView ArticlePubMed
- Kim S, Coulombe PA: Intermediate filament scaffolds fulfill mechanical, organizational, and signaling functions in the cytoplasm. Genes Dev. 2007, 21: 1581-1597. 10.1101/gad.1552107.View ArticlePubMed
- Pekny M, Pekna M: Astrocyte intermediate filaments in CNS pathologies and regeneration. J Pathol. 2004, 204: 428-437. 10.1002/path.1645.View ArticlePubMed
- Pekny M, Nilsson M: Astrocyte activation and reactive gliosis. Glia. 2005, 50: 427-434. 10.1002/glia.20207.View ArticlePubMed
- Eliasson C, Sahlgren C, Berthold C, Stakeberg J, Celis J, Betsholtz C, Eriksson J, Pekny M: Intermediate filament protein partnership in astrocytes. J Biol Chem. 1999, 274: 23996-24006. 10.1074/jbc.274.34.23996.View ArticlePubMed
- Potokar M, Kreft M, Li L, Daniel Andersson J, Pangrsic T, Chowdhury H, Pekny M, Zorec R: Cytoskeleton and vesicle mobility in astrocytes. Traffic. 2007, 8: 12-20. 10.1111/j.1600-0854.2006.00509.x.View ArticlePubMed
- Potokar M, Stenovec M, Gabrijel M, Li L, Kreft M, Grilc S, Pekny M, Zorec R: Intermediate filaments attenuate stimulation-dependent mobility of endosomes/lysosomes in astrocytes. Glia. 2010, 58: 1208-1219.PubMed
- Schwartz J, Wilson D: Preparation and characterization of type 1 astrocytes cultured from adult rat cortex, cerebellum, and striatum. Glia. 1992, 5: 75-80. 10.1002/glia.440050111.View ArticlePubMed
- Pekny M, Levéen P, Pekna M, Eliasson C, Berthold C, Westermark B, Betsholtz C: Mice lacking glial fibrillary acidic protein display astrocytes devoid of intermediate filaments but develop and reproduce normally. EMBO J. 1995, 14: 1590-1598.PubMed CentralPubMed
- Potokar M, Kreft M, Pangrsic T, Zorec R: Vesicle mobility studied in cultured astrocytes. Biochem Biophys Res Commun. 2005, 329: 678-683. 10.1016/j.bbrc.2005.02.030.View ArticlePubMed
- Pekny M, Johansson C, Eliasson C, Stakeberg J, Wallén A, Perlmann T, Lendahl U, Betsholtz C, Berthold C, Frisén J: Abnormal reaction to central nervous system injury in mice lacking glial fibrillary acidic protein and vimentin. J Cell Biol. 1999, 145: 503-514. 10.1083/jcb.145.3.503.PubMed CentralView ArticlePubMed
- Colucci-Guyon E, Portier M, Dunia I, Paulin D, Pournin S, Babinet C: Mice lacking vimentin develop and reproduce without an obvious phenotype. Cell. 1994, 79: 679-694. 10.1016/0092-8674(94)90553-3.View ArticlePubMed
- Bunbury A, Potolicchio I, Maitra R, Santambrogio L: Functional analysis of monocyte MHC class II compartments. FASEB J. 2009, 23: 164-171. 10.1096/fj.08-109439.PubMed CentralView ArticlePubMed
- van Niel G, Wubbolts R, Stoorvogel W: Endosomal sorting of MHC class II determines antigen presentation by dendritic cells. Curr Opin Cell Biol. 2008, 20: 437-444. 10.1016/j.ceb.2008.05.011.View ArticlePubMed
- Jaiswal JK, Andrews NW, Simon SM: Membrane proximal lysosomes are the major vesicles responsible for calcium-dependent exocytosis in nonsecretory cells. J Cell Biol. 2002, 159: 625-635. 10.1083/jcb.200208154.PubMed CentralView ArticlePubMed
- Gabrijel M, Kreft M, Zorec R: Monitoring lysosomal fusion in electrofused hybridoma cells. Biochim Biophys Acta. 2008, 1778: 483-490. 10.1016/j.bbamem.2007.10.013.View ArticlePubMed
- Jaiswal JK, Fix M, Takano T, Nedergaard M, Simon SM: Resolving vesicle fusion from lysis to monitor calcium-triggered lysosomal exocytosis in astrocytes. Proc Natl Acad Sci U S A. 2007, 104: 14151-14156. 10.1073/pnas.0704935104.PubMed CentralView ArticlePubMed
- Li D, Ropert N, Koulakoff A, Giaume C, Oheim M: Lysosomes are the major vesicular compartment undergoing Ca2 + -regulated exocytosis from cortical astrocytes. J Neurosci. 2008, 28: 7648-7658. 10.1523/JNEUROSCI.0744-08.2008.View ArticlePubMed
- Jiang M, Chen G: Ca2+ regulation of dynamin-independent endocytosis in cortical astrocytes. J Neurosci. 2009, 29: 8063-8074. 10.1523/JNEUROSCI.6139-08.2009.View ArticlePubMed
- Bennett MR, Farnell L, Gibson WG: A quantitative model of purinergic junctional transmission of calcium waves in astrocyte networks. Biophys J. 2005, 89: 2235-2250. 10.1529/biophysj.105.062968.PubMed CentralView ArticlePubMed
- Stenovec M, Kreft M, Grilc S, Potokar M, Kreft M, Pangrsic T, Zorec R: Ca2+-dependent mobility of vesicles capturing anti-VGLUT1 antibodies. Exp Cell Res. 2007, 313: 3809-3818. 10.1016/j.yexcr.2007.08.020.View ArticlePubMed
- Li L, Lundkvist A, Andersson D, Wilhelmsson U, Nagai N, Pardo AC, Nodin C, Ståhlberg A, Aprico K, Larsson K, Yabe T, Moons L, Fotheringham A, Davies I, Carmeliet P, Schwartz JP, Pekna M, Kubista M, Blomstrand F, Maragakis N, Nilsson M, Pekny M: Protective role of reactive astrocytes in brain ischemia. J Cereb Blood Flow Metab. 2008, 28: 468-481. 10.1038/sj.jcbfm.9600546.View ArticlePubMed
- Nieminen M, Henttinen T, Merinen M, Marttila-Ichihara F, Eriksson JE, Jalkanen S: Vimentin function in lymphocyte adhesion and transcellular migration. Nat Cell Biol. 2006, 8: 156-162. 10.1038/ncb1355.View ArticlePubMed
- Perlson E, Hanz S, Ben-Yaakov K, Segal-Ruder Y, Seger R, Fainzilber M: Vimentin-dependent spatial translocation of an activated MAP kinase in injured nerve. Neuron. 2005, 45: 715-726. 10.1016/j.neuron.2005.01.023.View ArticlePubMed
- Styers ML, Salazar G, Love R, Peden AA, Kowalczyk AP, Faundez V: The endo-lysosomal sorting machinery interacts with the intermediate filament cytoskeleton. Mol Biol Cell. 2004, 15: 5369-5382. 10.1091/mbc.E04-03-0272.PubMed CentralView ArticlePubMed
- Wagner OI, Lifshitz J, Janmey PA, Linden M, McIntosh TK, Leterrier JF: Mechanisms of mitochondria-neurofilament interactions. J Neurosci. 2003, 23: 9046-9058.PubMed
- Gao YS, Vrielink A, MacKenzie R, Sztul E: A novel type of regulation of the vimentin intermediate filament cytoskeleton by a Golgi protein. Eur J Cell Biol. 2002, 81: 391-401. 10.1078/0171-9335-00260.View ArticlePubMed
- Chang L, Barlan K, Chou YH, Grin B, Lakonishok M, Serpinskaya AS, Shumaker DK, Herrmann H, Gelfand VI, Goldman RD: The dynamic properties of intermediate filaments during organelle transport. J Cell Sci. 2009, 122: 2914-2923. 10.1242/jcs.046789.PubMed CentralView ArticlePubMed
- Stenovec M, Milošević M, Petrušić V, Potokar M, Stević Z, Prebil M, Kreft M, Trkov S, Andjus PR, Zorec R: Amyotrophic lateral sclerosis immunoglobulins G enhance the mobility of Lysotracker-labelled vesicles in cultured rat astrocytes. Acta Physiol (Oxf). 2011, 203: 457-471. 10.1111/j.1748-1716.2011.02337.x.View Article
- Chow A, Toomre D, Garrett W, Mellman I: Dendritic cell maturation triggers retrograde MHC class II transport from lysosomes to the plasma membrane. Nature. 2002, 418: 988-994. 10.1038/nature01006.View ArticlePubMed
- Boes M, Cerny J, Massol R, Op den Brouw M, Kirchhausen T, Chen J, Ploegh HL: T-cell engagement of dendritic cells rapidly rearranges MHC class II transport. Nature. 2002, 418: 983-988. 10.1038/nature01004.View ArticlePubMed
- Helfand BT, Chang L, Goldman RD: Intermediate filaments are dynamic and motile elements of cellular architecture. J Cell Sci. 2004, 117: 133-141. 10.1242/jcs.00936.View ArticlePubMed
- Chang L, Goldman RD: Intermediate filaments mediate cytoskeletal crosstalk. Nat Rev Mol Cell Biol. 2004, 5: 601-613. 10.1038/nrm1438.View ArticlePubMed
- Junyent F, De Lemos L, Utrera J, Paco S, Aguado F, Camins A, Pallàs M, Romero R, Auladell C: Content and traffic of taurine in hippocampal reactive astrocytes. Hippocampus. 2011, 21: 185-197. 10.1002/hipo.20739.View ArticlePubMed
- Wubbolts R, Fernandez-Borja M, Oomen L, Verwoerd D, Janssen H, Calafat J, Tulp A, Dusseljee S, Neefjes J: Direct vesicular transport of MHC class II molecules from lysosomal structures to the cell surface. J Cell Biol. 1996, 135: 611-622. 10.1083/jcb.135.3.611.View ArticlePubMed
- Zhang Z, Chen G, Zhou W, Song A, Xu T, Luo Q, Wang W, Gu XS, Duan S: Regulated ATP release from astrocytes through lysosome exocytosis. Nat Cell Biol. 2007, 9: 945-953. 10.1038/ncb1620.View ArticlePubMed
- Vardjan N, Stenovec M, Jorgacevski J, Kreft M, Zorec R: Subnanometer fusion pores in spontaneous exocytosis of peptidergic vesicles. J Neurosci. 2007, 27: 4737-4746. 10.1523/JNEUROSCI.0351-07.2007.View ArticlePubMed
- Harata N, Aravanis A, Tsien R: Kiss-and-run and full-collapse fusion as modes of exo-endocytosis in neurosecretion. J Neurochem. 2006, 97: 1546-1570. 10.1111/j.1471-4159.2006.03987.x.View ArticlePubMed
- Luzio JP, Pryor PR, Bright NA: Lysosomes: fusion and function. Nat Rev Mol Cell Biol. 2007, 8: 622-632. 10.1038/nrm2217.View ArticlePubMed
- Potokar M, Stenovec M, Kreft M, Kreft M, Zorec R: Stimulation inhibits the mobility of recycling peptidergic vesicles in astrocytes. Glia. 2008, 56: 135-144. 10.1002/glia.20597.View ArticlePubMed
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.