Structured Review

JEOL 100 electron microscope
Effect of ISS1 deletion on maturation of CPY (A) and Gas1p (B). After pulse-chase analysis of CPY or Gas1p, the intensities of radioactive bands were quantified with a PhosphorImager (Molecular Dynamics). Maturation was calculated as follows: CPY maturation (%) = m/(p1 + m) × <t>100;</t> Gas1p maturation (%) = m/(p + m) × 100, where p indicates precursor and m indicates mature form. (○) ISS1 strain; (●) iss1 null strain.
100 Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 88/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Sec24p and Iss1p Function Interchangeably in Transport Vesicle Formation from the Endoplasmic Reticulum in Saccharomyces cerevisiae"

Article Title: Sec24p and Iss1p Function Interchangeably in Transport Vesicle Formation from the Endoplasmic Reticulum in Saccharomyces cerevisiae

Journal: Molecular Biology of the Cell

doi:

Effect of ISS1 deletion on maturation of CPY (A) and Gas1p (B). After pulse-chase analysis of CPY or Gas1p, the intensities of radioactive bands were quantified with a PhosphorImager (Molecular Dynamics). Maturation was calculated as follows: CPY maturation (%) = m/(p1 + m) × 100; Gas1p maturation (%) = m/(p + m) × 100, where p indicates precursor and m indicates mature form. (○) ISS1 strain; (●) iss1 null strain.
Figure Legend Snippet: Effect of ISS1 deletion on maturation of CPY (A) and Gas1p (B). After pulse-chase analysis of CPY or Gas1p, the intensities of radioactive bands were quantified with a PhosphorImager (Molecular Dynamics). Maturation was calculated as follows: CPY maturation (%) = m/(p1 + m) × 100; Gas1p maturation (%) = m/(p + m) × 100, where p indicates precursor and m indicates mature form. (○) ISS1 strain; (●) iss1 null strain.

Techniques Used: Pulse Chase

2) Product Images from "Thermal adaptation of mesophilic and thermophilic FtsZ assembly by modulation of the critical concentration"

Article Title: Thermal adaptation of mesophilic and thermophilic FtsZ assembly by modulation of the critical concentration

Journal: PLoS ONE

doi: 10.1371/journal.pone.0185707

The morphology of FtsZ polymers was characterized by negative-stain transmission electron microscopy. Top row, 10 μM EcFtsZ was polymerized at 10, 20 and 30°C (A-C). Bottom row, 10 μM MjFtsZ was polymerized at 40, 60 and 80°C (D–F). The arrowheads in D point to examples of short and curved polymers. The scale bars represent 100 and 500 nm for the top and bottom rows, respectively (black bars).
Figure Legend Snippet: The morphology of FtsZ polymers was characterized by negative-stain transmission electron microscopy. Top row, 10 μM EcFtsZ was polymerized at 10, 20 and 30°C (A-C). Bottom row, 10 μM MjFtsZ was polymerized at 40, 60 and 80°C (D–F). The arrowheads in D point to examples of short and curved polymers. The scale bars represent 100 and 500 nm for the top and bottom rows, respectively (black bars).

Techniques Used: Staining, Transmission Assay, Electron Microscopy

3) Product Images from "The Interaction of Neurofilaments with the Microtubule Motor Cytoplasmic Dynein"

Article Title: The Interaction of Neurofilaments with the Microtubule Motor Cytoplasmic Dynein

Journal: Molecular Biology of the Cell

doi: 10.1091/mbc.E04-05-0401

AFM images of cytoplasmic dynein/dynactin complex bound to neurofilaments. (a) Dynein/dynactin decorate neurofilaments along their lengths, often forming thin projections of length 50-100 nm extending from the neurofilament core. (b) The binding of dynein/dynactin
Figure Legend Snippet: AFM images of cytoplasmic dynein/dynactin complex bound to neurofilaments. (a) Dynein/dynactin decorate neurofilaments along their lengths, often forming thin projections of length 50-100 nm extending from the neurofilament core. (b) The binding of dynein/dynactin

Techniques Used: Binding Assay

Related Articles

Staining:

Article Title: The Interaction of Neurofilaments with the Microtubule Motor Cytoplasmic Dynein
Article Snippet: .. Samples were loaded on glow-discharged formvar-carbon-coated 300 mesh copper grids, stained by 1% uranyl acetate in H2 O for 20 s, dried in air after side adsorption of the drop, and observed in a Jeol 100 electron microscope (Peabody, MA) at 80 kV. .. We used the LexA yeast two-hybrid system to screen a random-primed human brain library (Stratagene, La Jolla, CA) for proteins interacting with DIC.

Article Title: Direct and indirect interactions of the dopamine D3 receptor with glutamate pathways: implications for the treatment of schizophrenia
Article Snippet: .. Ultrathin sections were mounted on mesh grids, stained with 0.4 % lead citrate and 4.0 % uranyl acetate, and finally analyzed and photographed on a JEOL 100 electron microscope. .. Treatments for chronic NMDA receptor blockade For chronic NMDA receptor blockade, dizocilpine was continuously infused for 7 days via osmotic minipumps (Model 1007D, Alzet Corp.) from day 0 to day 7.

Article Title: Sec24p and Iss1p Function Interchangeably in Transport Vesicle Formation from the Endoplasmic Reticulum in Saccharomyces cerevisiae
Article Snippet: .. Thin sections were stained with lead citrate and viewed in a JEOL 100 electron microscope ( JEOL , Tokyo, Japan). ..

Article Title: Prion protein expression in different species: Analysis with a panel of new mAbs
Article Snippet: .. Ultrathin sections were cut with a ultramicrotome (Ultracut FC4; Reichert), stained with uranyl acetate and lead citrate, and examined with a JEOL 100 electron microscope. ..

Microscopy:

Article Title: Complement Depletion Reduces Macrophage Infiltration and Activation during Wallerian Degeneration and Axonal Regeneration
Article Snippet: .. Ultrathin sections were cut on a Reichert–Jung Ultracut E microtome and viewed on a JEOL 100 electron microscope. .. The area of each field was measured with a digitizing tablet (Microcomputer Imaging Device, IBM; software version 4.20, Imaging Research, St. Catherine’s, Ontario, Canada), which converted screen pixels to surface area; a standard area was chosen so that identical volumes from each nerve were examined.

Article Title: The Interaction of Neurofilaments with the Microtubule Motor Cytoplasmic Dynein
Article Snippet: .. Samples were loaded on glow-discharged formvar-carbon-coated 300 mesh copper grids, stained by 1% uranyl acetate in H2 O for 20 s, dried in air after side adsorption of the drop, and observed in a Jeol 100 electron microscope (Peabody, MA) at 80 kV. .. We used the LexA yeast two-hybrid system to screen a random-primed human brain library (Stratagene, La Jolla, CA) for proteins interacting with DIC.

Article Title: Direct and indirect interactions of the dopamine D3 receptor with glutamate pathways: implications for the treatment of schizophrenia
Article Snippet: .. Ultrathin sections were mounted on mesh grids, stained with 0.4 % lead citrate and 4.0 % uranyl acetate, and finally analyzed and photographed on a JEOL 100 electron microscope. .. Treatments for chronic NMDA receptor blockade For chronic NMDA receptor blockade, dizocilpine was continuously infused for 7 days via osmotic minipumps (Model 1007D, Alzet Corp.) from day 0 to day 7.

Article Title: Induction of Autophagy and Inhibition of Melanoma Growth In Vitro and In Vivo by Hyperactivation of Oncogenic BRAF
Article Snippet: .. Ultrathin sections were observed and photographed using the Jeol 100 electron microscope (Tokyo, Japan) at the UW Electron Microscope Facility. .. Athymic Ncr-nu/nu mice (4–6 weeks, male and female) were purchased from the National Cancer Institute (NCI, Frederick, MD).

Article Title: Thermal adaptation of mesophilic and thermophilic FtsZ assembly by modulation of the critical concentration
Article Snippet: .. The grids were visualized in a Phillips Tecnai 12 Bio Twin Electron Microscope at 49,000X (for EcFtsZ), or with a Jeol 100 electron microscope at 20,000X (for MjFtsZ). .. The polymer widths were measured using ImageJ software ( http://imagej.nih.gov/ij/ ).

Article Title: Complement Depletion Reduces Macrophage Infiltration and Activation during Wallerian Degeneration and Axonal Regeneration
Article Snippet: .. Ultrathin sections were cut on a Reichert–Jung Ultracut E microtome and viewed on a JEOL 100 electron microscope. .. Macrophage counts were performed at 200× magnification by counting all positively stained cells that contained a visible nucleus in three randomly chosen fields in each degenerating nerve segment.

Article Title: Prion protein expression in different species: Analysis with a panel of new mAbs
Article Snippet: .. Ultrathin sections were cut with a ultramicrotome (Ultracut FC4; Reichert), stained with uranyl acetate and lead citrate, and examined with a JEOL 100 electron microscope. ..

Adsorption:

Article Title: The Interaction of Neurofilaments with the Microtubule Motor Cytoplasmic Dynein
Article Snippet: .. Samples were loaded on glow-discharged formvar-carbon-coated 300 mesh copper grids, stained by 1% uranyl acetate in H2 O for 20 s, dried in air after side adsorption of the drop, and observed in a Jeol 100 electron microscope (Peabody, MA) at 80 kV. .. We used the LexA yeast two-hybrid system to screen a random-primed human brain library (Stratagene, La Jolla, CA) for proteins interacting with DIC.

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    JEOL 100 cx electron microscope
    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; <t>100</t> mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.
    100 Cx Electron Microscope, supplied by JEOL, used in various techniques. Bioz Stars score: 89/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 89 stars, based on 122 article reviews
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    The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; 100 mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.

    Journal: Genes & Development

    Article Title: members only encodes a Drosophila nucleoporin required for Rel protein import and immune response activation

    doi:

    Figure Lengend Snippet: The expression and phenotypes of mbo are cell-specific. ( A,B ) Confocal images of a stage-16 embryo carrying one copy of the mbo–lacZ reporter stained with anti-β-galactosidase (red) and mAb 2A12 to visualize the tracheal lumen (green). ( A ) lacZ expression is detected in fusion cells (asterisks) of the dorsal branches (DB) forming the dorsal anastomosis (DA). mbo–lacZ is not expressed in the stalk cells of the DB or the cells extending terminal branches (TB). ( B ) mbo–lacZ is expressed in the fusion cells (asterisks) of the dorsal trunk (DT) but not in the stalk cells of the DB or in the transverse connective (TC). The DB is out of focus; its position is drawn with a broken line. Bars, 5 μm ( A ); 2 μm ( B ). ( C,D ) In situ hybridization to mbo mRNA in third instar larval CNS. In wild type ( C ) mbo is expressed in proliferating cells of the nerve cord (brackets), in the optic lobes (OL) of the brain, and the imaginal discs shown attached to the lobes. mbo RNA is not detectable in the CNS of mbo mutants ( D ), and the size of the CNS is reduced. Bars in C and D ; 100 mm. ( E,K ) Dorsal anastomoses in late stage-16 wild-type (asterisk in E ) and mbo mutant ( K ) embryos. In mbo mutant embryos, 20% of the dorsal branches fail to connect (arrowhead in K ) Bar, 10 μm. ( F,L ) Dorsal anastomoses in third instar wild-type (asterisk in F ) and mbo mutant ( L ) larvae. In mutants the DBs are disconnected (arrowhead), but terminal branching is not affected (arrows in F,L ). Bar, 50 μm. ( G,M ) Dorsal anastomoses in stage-16 embryos carrying one copy of the esg–lacZ marker. esg–lacZ is expressed in the fusion cells of both wild type ( G ) and mbo mutants ( M ). Bars in G and M , 2 μm. ( H,N ) Segments of the dorsal trunks of wild-type and mbo third instar larvae. In mutants the cuticular lining of the dorsal trunks is disrupted at the positions of the fusion junctions (arrowheads in N ) compared to junctions in the wild type (asterisks in H ). Bar, 50 μm ( I–P ) Dnup88 expression in larval fat body detected with the antiserum against the amino-terminal part of the protein. Nuclear staining is detected in wild-type larvae ( I ) but absent in mbo mutants ( O ). The nuclei are visualized by DAPI staining in the adjacent panels J and P . Bar in I,J,O , and P , 40 μm.

    Article Snippet: Wild-type and mbo mutant larvae were prepared for EM as described ( ) and examined with a Jeol 100 CX electron microscope.

    Techniques: Expressing, Staining, In Situ Hybridization, Mutagenesis, Marker

    mbo is not required for mRNA export. ( A–C ) In situ hybridization to lacZ RNA in wild-type and mbo larvae carrying the hs–GAL4 and UAS–lacZNLS transgenes. The lacZ RNA is detected in the proventriculus of wild-type ( B ) and mbo mutant ( C ) larvae after heat shock and does not accumulate in the nucleus (arrowheads). The dark spot inside each nucleus is likely to correlate with the site of transcription. lacZ expression is reduced in some of the cells of mbo mutants (arrows). Bar, 10 μm. ( D–F ) Heat shock-induced expression of Hdc protein in wild-type and mbo mutants. Fat bodies from untreated wild-type ( D ) and heat-shocked wild-type ( E ) and mbo ( F ) larvae carrying the hs–hdc transgene were stained with an antibody against the Hdc protein. Bar, 50 μm. ( G ) Electron micrograph of a section through the lymph gland of an mbo larva. In this tangential section, several NPCs (arrow) can be identified in the space between the cytoplasm (Cyt) and the nucleus (Nuc). Their distribution and morphology are indistinguishable from wild type at this level. Bar, 100 nm.

    Journal: Genes & Development

    Article Title: members only encodes a Drosophila nucleoporin required for Rel protein import and immune response activation

    doi:

    Figure Lengend Snippet: mbo is not required for mRNA export. ( A–C ) In situ hybridization to lacZ RNA in wild-type and mbo larvae carrying the hs–GAL4 and UAS–lacZNLS transgenes. The lacZ RNA is detected in the proventriculus of wild-type ( B ) and mbo mutant ( C ) larvae after heat shock and does not accumulate in the nucleus (arrowheads). The dark spot inside each nucleus is likely to correlate with the site of transcription. lacZ expression is reduced in some of the cells of mbo mutants (arrows). Bar, 10 μm. ( D–F ) Heat shock-induced expression of Hdc protein in wild-type and mbo mutants. Fat bodies from untreated wild-type ( D ) and heat-shocked wild-type ( E ) and mbo ( F ) larvae carrying the hs–hdc transgene were stained with an antibody against the Hdc protein. Bar, 50 μm. ( G ) Electron micrograph of a section through the lymph gland of an mbo larva. In this tangential section, several NPCs (arrow) can be identified in the space between the cytoplasm (Cyt) and the nucleus (Nuc). Their distribution and morphology are indistinguishable from wild type at this level. Bar, 100 nm.

    Article Snippet: Wild-type and mbo mutant larvae were prepared for EM as described ( ) and examined with a Jeol 100 CX electron microscope.

    Techniques: In Situ Hybridization, Mutagenesis, Expressing, Staining

    Ultrastructural colocalization of peripherin and NFL on the same neurofilament in sciatic nerve by pre-embedding immuno-EM Paraformaldehyde-fixed samples were incubated with rabbit anti-peripherin and mouse anti-NFL antibodies and probed with goat anti-rabbit IgG and goat anti-mouse IgG conjugated to 0.6nm gold beads. Single (for anti-mouse IgG)- or double (for anti-rabbit IgG)-silver enhancement of gold particles resulted in irregular-shaped electron-dense particles that could be distinguished by their size. As expected, for the immunodetection of peripherin and NFL in normal mice ( A ), linear arrays of two sizes of gold particles (large for peripherin and samll for NFL) decorate most 10-nm filaments in the axon and negligible numbers are detected in peripherin knockout mice ( E ). Higher magnification shows that gold particles of two sizes overlie a single filament in the background ( B, C, D ). Arrows point to small particles (NFL) and arrowheads to large ones (peripherin). Scale bars, 100 nm in A ; 60 nm in B ; 40 nm in C ; 50 nm in D ; 200 nm in E .

    Journal: The Journal of Neuroscience

    Article Title: Peripherin Is a Subunit of Peripheral Nerve Neurofilaments: Implications for Differential Vulnerability of CNS and PNS Axons

    doi: 10.1523/JNEUROSCI.1081-12.2012

    Figure Lengend Snippet: Ultrastructural colocalization of peripherin and NFL on the same neurofilament in sciatic nerve by pre-embedding immuno-EM Paraformaldehyde-fixed samples were incubated with rabbit anti-peripherin and mouse anti-NFL antibodies and probed with goat anti-rabbit IgG and goat anti-mouse IgG conjugated to 0.6nm gold beads. Single (for anti-mouse IgG)- or double (for anti-rabbit IgG)-silver enhancement of gold particles resulted in irregular-shaped electron-dense particles that could be distinguished by their size. As expected, for the immunodetection of peripherin and NFL in normal mice ( A ), linear arrays of two sizes of gold particles (large for peripherin and samll for NFL) decorate most 10-nm filaments in the axon and negligible numbers are detected in peripherin knockout mice ( E ). Higher magnification shows that gold particles of two sizes overlie a single filament in the background ( B, C, D ). Arrows point to small particles (NFL) and arrowheads to large ones (peripherin). Scale bars, 100 nm in A ; 60 nm in B ; 40 nm in C ; 50 nm in D ; 200 nm in E .

    Article Snippet: The grids were photographed on a JEOL 100 cx electron microscope operated at 80 kV.

    Techniques: Incubation, Immunodetection, Mouse Assay, Knock-Out

    Knockdown of SR-BI/CLA-1 in Caco-2/TC7 cells impairs PPM-induced ERK1/2 phosphorylation and apoB chase. (A) Caco-2/TC7 Cell populations 63 and 64, expressing lentiviral shRNA 63 and 64 respectively, were analyzed at passage 4 after transfection in the absence of PPM or after 10 min of PPM supply. Cell lysates were analyzed by immunoblot with antibodies against SR-BI/CLA-1 and E-cadherin (E-cadh, used as loading control). The lower panel shows the level of SR-BI/CLA-1 expression normalized to the level of E-cadherin expression set at 100% for control Caco-2/TC7 cells. Results are from two independent sets of experiments. (B) Cell populations 63 and 64 were cultured on semi-permeable filters and incubated in the absence or presence of PPM or IPM in the apical compartment for the indicated times. An early (63E) and a late (63L) passage (corresponding respectively to passage 6 and 28 after transfection) of Cell population 63 were compared to Cell population 64 at passage 28. Cell lysates were analyzed in SR-BI/CLA-1 and phospho-ERK1/2 (P-ERK) immunoblots. Total ERK (ERK) and E-cadherin (E-cadh) were used as loading controls. Lower panel, the ratio of P-ERK expression normalized to total ERK expression in PPM-treated cells versus IPM-treated cells, set at 100% for Cell population 64. Results show the means±SEM of three independent sets of experiments. *P

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: Knockdown of SR-BI/CLA-1 in Caco-2/TC7 cells impairs PPM-induced ERK1/2 phosphorylation and apoB chase. (A) Caco-2/TC7 Cell populations 63 and 64, expressing lentiviral shRNA 63 and 64 respectively, were analyzed at passage 4 after transfection in the absence of PPM or after 10 min of PPM supply. Cell lysates were analyzed by immunoblot with antibodies against SR-BI/CLA-1 and E-cadherin (E-cadh, used as loading control). The lower panel shows the level of SR-BI/CLA-1 expression normalized to the level of E-cadherin expression set at 100% for control Caco-2/TC7 cells. Results are from two independent sets of experiments. (B) Cell populations 63 and 64 were cultured on semi-permeable filters and incubated in the absence or presence of PPM or IPM in the apical compartment for the indicated times. An early (63E) and a late (63L) passage (corresponding respectively to passage 6 and 28 after transfection) of Cell population 63 were compared to Cell population 64 at passage 28. Cell lysates were analyzed in SR-BI/CLA-1 and phospho-ERK1/2 (P-ERK) immunoblots. Total ERK (ERK) and E-cadherin (E-cadh) were used as loading controls. Lower panel, the ratio of P-ERK expression normalized to total ERK expression in PPM-treated cells versus IPM-treated cells, set at 100% for Cell population 64. Results show the means±SEM of three independent sets of experiments. *P

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Expressing, shRNA, Transfection, Cell Culture, Incubation, Western Blot

    Subcellular localization of SR-BI/CLA-1 after the supply of postprandial micelles. (A) Immunoelectron micrograph of SR-BI/CLA-1 in untreated differentiated Caco-2/TC7 cells. MV, microvilli; TW, terminal web (bar, 0.5 µm). Note the significant amount of intracellular trafficking SR-BI/CLA-1 in addition to its main apical localization (arrowheads). (B) Immunolocalization of SR-BI/CLA-1 (green channel) and sucrase isomaltase (SI, red channel) in differentiated Caco-2/TC7 cells before (T0) and after 5, 10 and 15 min of apical PPM supply. Panels represent XY acquisitions at the apical level (bar, 10 µm). Arrowheads show clusters of SR-BI/CLA-1. (C) Immunolocalization of SR-BI/CLA-1 in differentiated Caco-2/TC7 cells in the absence (control) or presence of PPM or IPM for 20 min (bar, 20 µm). Arrowheads show clusters of SR-BI/CLA-1 (D) Immunoelectron micrograph of SR-BI/CLA-1 in Caco-2/TC7 cells supplied with PPM (MV, microvilli). Arrowheads indicate SR-BI/CLA-1 clusters (bar, 100 nm). (E) Cell surface biotinylation assay for apical SR-BI/CLA-1. Caco-2/TC7 cells were cultured in the absence (0) or presence of PPM for the indicated times. Cells were then selectively labeled with non-permeant biotin at the apical (left panel) or basal surface (right panel). Biotinylated fractions were obtained as described in Material and Methods . Total cell lysates (total), apical and basal biotinylated fractions (left and right panel respectively) and non-apical fractions (non-apical) were analyzed in immunoblots of SR-BI/CLA-1, E-cadherin being used as a basolateral membrane marker.

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: Subcellular localization of SR-BI/CLA-1 after the supply of postprandial micelles. (A) Immunoelectron micrograph of SR-BI/CLA-1 in untreated differentiated Caco-2/TC7 cells. MV, microvilli; TW, terminal web (bar, 0.5 µm). Note the significant amount of intracellular trafficking SR-BI/CLA-1 in addition to its main apical localization (arrowheads). (B) Immunolocalization of SR-BI/CLA-1 (green channel) and sucrase isomaltase (SI, red channel) in differentiated Caco-2/TC7 cells before (T0) and after 5, 10 and 15 min of apical PPM supply. Panels represent XY acquisitions at the apical level (bar, 10 µm). Arrowheads show clusters of SR-BI/CLA-1. (C) Immunolocalization of SR-BI/CLA-1 in differentiated Caco-2/TC7 cells in the absence (control) or presence of PPM or IPM for 20 min (bar, 20 µm). Arrowheads show clusters of SR-BI/CLA-1 (D) Immunoelectron micrograph of SR-BI/CLA-1 in Caco-2/TC7 cells supplied with PPM (MV, microvilli). Arrowheads indicate SR-BI/CLA-1 clusters (bar, 100 nm). (E) Cell surface biotinylation assay for apical SR-BI/CLA-1. Caco-2/TC7 cells were cultured in the absence (0) or presence of PPM for the indicated times. Cells were then selectively labeled with non-permeant biotin at the apical (left panel) or basal surface (right panel). Biotinylated fractions were obtained as described in Material and Methods . Total cell lysates (total), apical and basal biotinylated fractions (left and right panel respectively) and non-apical fractions (non-apical) were analyzed in immunoblots of SR-BI/CLA-1, E-cadherin being used as a basolateral membrane marker.

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Cell Surface Biotinylation Assay, Cell Culture, Labeling, Western Blot, Marker

    PPM supply induces movement of SR-BI/CLA-1 towards raft microdomains. (A) Caco-2/TC7 cells were harvested in the presence of Triton X-100 and the lysate fractionated on a 5–40% sucrose gradient. Eleven fractions were collected for immunoblots of SR-BI/CLA-1, EEA1 (early endosome antigen 1) and flottilin-1 (raft marker). (B) Caco-2/TC7 cells were cultured in the absence (control) or presence of PPM or IPM for 10 min and then harvested in the presence of Triton X-100. Cell lysates were applied to a 5–40% sucrose gradient and eleven fractions collected. Fractions 3 to 8 were analyzed by immunoblotting with antibodies against SR-BI/CLA-1 (left panel) and flottilin-1 (right panel). (C) Immunolocalization of SR-BI/CLA-1 and alkaline phosphatase (PLAP, used as raft marker) in the brush border of Caco-2/TC7 cells supplied with PPM. SR-BI/CLA-1 is labelled with anti-rabbit immunoglobulin-gold complexes (18-nm particles) and PLAP with anti-sheep immunoglobulin-gold complexes (12-nm particles). MV, microvilli; bar, 100 nm.

    Journal: PLoS ONE

    Article Title: Sensing of Dietary Lipids by Enterocytes: A New Role for SR-BI/CLA-1

    doi: 10.1371/journal.pone.0004278

    Figure Lengend Snippet: PPM supply induces movement of SR-BI/CLA-1 towards raft microdomains. (A) Caco-2/TC7 cells were harvested in the presence of Triton X-100 and the lysate fractionated on a 5–40% sucrose gradient. Eleven fractions were collected for immunoblots of SR-BI/CLA-1, EEA1 (early endosome antigen 1) and flottilin-1 (raft marker). (B) Caco-2/TC7 cells were cultured in the absence (control) or presence of PPM or IPM for 10 min and then harvested in the presence of Triton X-100. Cell lysates were applied to a 5–40% sucrose gradient and eleven fractions collected. Fractions 3 to 8 were analyzed by immunoblotting with antibodies against SR-BI/CLA-1 (left panel) and flottilin-1 (right panel). (C) Immunolocalization of SR-BI/CLA-1 and alkaline phosphatase (PLAP, used as raft marker) in the brush border of Caco-2/TC7 cells supplied with PPM. SR-BI/CLA-1 is labelled with anti-rabbit immunoglobulin-gold complexes (18-nm particles) and PLAP with anti-sheep immunoglobulin-gold complexes (12-nm particles). MV, microvilli; bar, 100 nm.

    Article Snippet: Sections were analyzed in a Jeol 100 CX II electron microscope.

    Techniques: Western Blot, Marker, Cell Culture

    Immunoreactive COX-2 expression in vasculature-associated cells in response to IL-1 versus LPS. Bright-field images of COX-2-ir cells associated with the vasculature in the forebrain ( top ) and medulla ( bottom ) from rats killed 4 hr after intravenous injection of IL-1 (1.87 μg, left ) or LPS (100 μg/kg, right ). As with IL-1, at 4 hr after LPS treatment, a clear increase in the number and staining intensity of COX-2-positive cells is seen within perivascular regions throughout the brain. However, the predominant cell types manifesting enzyme expression after each treatment are morphologically distinct. COX-2-positive polygonal/multipolar cells ( open arrows ) are seen in response to each treatment and exclusively in material from IL-1-treated animals. COX-2-positive round cells ( arrowheads ) are evident only in rats treated with LPS. Scale bar, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Immunoreactive COX-2 expression in vasculature-associated cells in response to IL-1 versus LPS. Bright-field images of COX-2-ir cells associated with the vasculature in the forebrain ( top ) and medulla ( bottom ) from rats killed 4 hr after intravenous injection of IL-1 (1.87 μg, left ) or LPS (100 μg/kg, right ). As with IL-1, at 4 hr after LPS treatment, a clear increase in the number and staining intensity of COX-2-positive cells is seen within perivascular regions throughout the brain. However, the predominant cell types manifesting enzyme expression after each treatment are morphologically distinct. COX-2-positive polygonal/multipolar cells ( open arrows ) are seen in response to each treatment and exclusively in material from IL-1-treated animals. COX-2-positive round cells ( arrowheads ) are evident only in rats treated with LPS. Scale bar, 100 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Expressing, Injection, Staining

    LPS-induced COX-2 expression in vasculature-associated cells. SCLM images show dual immunostaining for COX-2 ( green , left ) and RECA-1, a marker for endothelial cells ( red , middle ), in blood vessels in the forebrain. Results of dual immunolabeling of material from rats challenged with 100 μg/kg LPS revealed that many round COX-2-ir cells coexpress the endothelial marker RECA-1 ( right panel ). Another population of COX-2-ir cells, polygonal or multipolar in form, stained positively for the ED2 antigen (data not shown) in LPS-treated rats. The yellow color in the merged image ( right ) represents a positive signal for both markers and is consistent with a perinuclear distribution of COX-2-ir in activated endothelial cells. Arrowhead indicates a COX-2- and RECA-1-positive cell. Arrow indicates a multipolar COX-2-positive cell that did not express RECA-1. Scale bar, 50 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: LPS-induced COX-2 expression in vasculature-associated cells. SCLM images show dual immunostaining for COX-2 ( green , left ) and RECA-1, a marker for endothelial cells ( red , middle ), in blood vessels in the forebrain. Results of dual immunolabeling of material from rats challenged with 100 μg/kg LPS revealed that many round COX-2-ir cells coexpress the endothelial marker RECA-1 ( right panel ). Another population of COX-2-ir cells, polygonal or multipolar in form, stained positively for the ED2 antigen (data not shown) in LPS-treated rats. The yellow color in the merged image ( right ) represents a positive signal for both markers and is consistent with a perinuclear distribution of COX-2-ir in activated endothelial cells. Arrowhead indicates a COX-2- and RECA-1-positive cell. Arrow indicates a multipolar COX-2-positive cell that did not express RECA-1. Scale bar, 50 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Expressing, Immunostaining, Marker, Immunolabeling, Staining

    Fine structure of LPS-sensitive vasculature-associated cells. Electron micrographs showing pre-embedding immunoperoxidase labeling for COX-2 in vasculature-associated cells in the forebrain of a rat treated with 100 μg/kg LPS. COX-2-ir is distributed diffusely within the cytoplasm of a cell ( top panel , dotted line ) that is not an integral component of the vascular wall, is segregated from the brain parenchyma by a basal lamina, and displays morphological features similar to ED2-positive perivascular cells. The bottom panels show examples of COX-2-ir within endothelial cells. Note the perinuclear distribution of the reaction product, consistent with the light-level appearance of COX-2-ir in this cell type. Arrows indicate positive labeling for COX-2. N , Nucleus; bl , basal lamina; EC , endothelial cell; bv , blood vessel. Scale bar, 1 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Fine structure of LPS-sensitive vasculature-associated cells. Electron micrographs showing pre-embedding immunoperoxidase labeling for COX-2 in vasculature-associated cells in the forebrain of a rat treated with 100 μg/kg LPS. COX-2-ir is distributed diffusely within the cytoplasm of a cell ( top panel , dotted line ) that is not an integral component of the vascular wall, is segregated from the brain parenchyma by a basal lamina, and displays morphological features similar to ED2-positive perivascular cells. The bottom panels show examples of COX-2-ir within endothelial cells. Note the perinuclear distribution of the reaction product, consistent with the light-level appearance of COX-2-ir in this cell type. Arrows indicate positive labeling for COX-2. N , Nucleus; bl , basal lamina; EC , endothelial cell; bv , blood vessel. Scale bar, 1 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Labeling

    Central prostaglandin synthesis blockade disrupts systemic IL-1-induced activation of the paraventricular nucleus and its aminergic afferents. Bright-field photomicrographs show IL-1-induced Fos-ir expression in the paraventricular nucleus ( PVH , top ) and the C1 region of the ventrolateral medulla ( VLM , bottom ) in rats pretreated by intracerebroventricular injection of vehicle ( left ) or indomethacin (10 μg/5 μl) ( right ). As reported previously, intravenous IL-1 (1.87 μg/kg) evokes a robust Fos response within the PVH and C1 regions. However, pretreatment with central infusion of indomethacin, a nonselective inhibitor of COX activity, results in a marked diminution of IL-1 effects at the levels of both medulla and hypothalamus. This finding supports the view that induced synthesis of prostaglandins within the brain is required for the activation of HPA control systems in response to systemic (intravenous) IL-1. Scale bar, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Central prostaglandin synthesis blockade disrupts systemic IL-1-induced activation of the paraventricular nucleus and its aminergic afferents. Bright-field photomicrographs show IL-1-induced Fos-ir expression in the paraventricular nucleus ( PVH , top ) and the C1 region of the ventrolateral medulla ( VLM , bottom ) in rats pretreated by intracerebroventricular injection of vehicle ( left ) or indomethacin (10 μg/5 μl) ( right ). As reported previously, intravenous IL-1 (1.87 μg/kg) evokes a robust Fos response within the PVH and C1 regions. However, pretreatment with central infusion of indomethacin, a nonselective inhibitor of COX activity, results in a marked diminution of IL-1 effects at the levels of both medulla and hypothalamus. This finding supports the view that induced synthesis of prostaglandins within the brain is required for the activation of HPA control systems in response to systemic (intravenous) IL-1. Scale bar, 100 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Activation Assay, Expressing, Injection, Activity Assay

    Vascular COX-2-ir induction as a function of IL-1 dose. Bright-field images show blood vessels stained for COX-2 ( top row ) or the PVN labeled for Fos-ir ( bottom row ), from rats given vehicle ( left panels ), 1.87 μg/kg ( middle panels ), or 30 μg/kg IL-1 ( right panels ). To provide an index of the strength of the stimulus, Fos-ir induction in the PVH seen in response to the same treatments is shown ( bottom row ). In vehicle-treated rats, few to no COX-2-ir cells are found in association with blood vessels ( top left ), and Fos expression is not detected within the PVH ( bottom left ). As documented previously, 1.87 μg/kg doses of IL-1 stimulate COX-2-expression within polygonal or multipolar cells presumed to conform to ED2-positive perivascular cells ( top , middle , open arrows ); moderate Fos-ir induction is localized principally to the medial parvocellular ( mp ) part of the PVH, with lesser involvement of the dorsal parvocellular ( dp )and posterior magnocellular ( pm ) subdivisions ( bottom , middle ). The 30 μg/kg IL-1 dose produces more robust Fos induction in the PVH ( bottom , right ), comparable to that seen in response to 2 μg/kg LPS. Nevertheless, only elements exhibiting perivascular cell morphology manifest COX-2-ir in response to the higher IL-1 dose ( top , right ). Scale bar, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Vascular COX-2-ir induction as a function of IL-1 dose. Bright-field images show blood vessels stained for COX-2 ( top row ) or the PVN labeled for Fos-ir ( bottom row ), from rats given vehicle ( left panels ), 1.87 μg/kg ( middle panels ), or 30 μg/kg IL-1 ( right panels ). To provide an index of the strength of the stimulus, Fos-ir induction in the PVH seen in response to the same treatments is shown ( bottom row ). In vehicle-treated rats, few to no COX-2-ir cells are found in association with blood vessels ( top left ), and Fos expression is not detected within the PVH ( bottom left ). As documented previously, 1.87 μg/kg doses of IL-1 stimulate COX-2-expression within polygonal or multipolar cells presumed to conform to ED2-positive perivascular cells ( top , middle , open arrows ); moderate Fos-ir induction is localized principally to the medial parvocellular ( mp ) part of the PVH, with lesser involvement of the dorsal parvocellular ( dp )and posterior magnocellular ( pm ) subdivisions ( bottom , middle ). The 30 μg/kg IL-1 dose produces more robust Fos induction in the PVH ( bottom , right ), comparable to that seen in response to 2 μg/kg LPS. Nevertheless, only elements exhibiting perivascular cell morphology manifest COX-2-ir in response to the higher IL-1 dose ( top , right ). Scale bar, 100 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Staining, Labeling, Expressing

    Strength and locus of COX-2 induction as a function of LPS dose. Bright-field images show vessels stained for COX-2 ( top row ) from rats given 0.1 μg/kg ( left ), 2 μg/kg ( middle ), or 100 μg/kg ( right ) LPS. In rats treated with 0.1 μg/kg LPS ( left ), only cells exhibiting perivascular cell morphology manifest COX-2-ir ( left , open arrows ). Even this low dose provokes significant activation of neurons within the PVH, especially its CRF-rich medial parvocellular ( mp ) subdivision. In response to the 2 μg/kg LPS dose ( middle ), both polygonal/multipolar ( open arrows ) and round-shaped cells ( closed arrowheads ) exhibit COX-2-ir, suggesting involvement of both perivascular and endothelial cells. Fos induction under this condition is marginally increased, with greater involvement of the dorsal parvocellular ( dp ) and posterior magnocellular ( pm ) aspects of the PVH. The 100 μg/kg LPS dose ( right ) also provokes COX-2-ir expression in both polygonal/multipolar ( open arrows ) and round ( arrowheads ) cells, whose number and staining intensity are enhanced. Fos induction in the PVH is most robust under this condition and distributed uniformly throughout all subregions of the nucleus. Scale bar, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Strength and locus of COX-2 induction as a function of LPS dose. Bright-field images show vessels stained for COX-2 ( top row ) from rats given 0.1 μg/kg ( left ), 2 μg/kg ( middle ), or 100 μg/kg ( right ) LPS. In rats treated with 0.1 μg/kg LPS ( left ), only cells exhibiting perivascular cell morphology manifest COX-2-ir ( left , open arrows ). Even this low dose provokes significant activation of neurons within the PVH, especially its CRF-rich medial parvocellular ( mp ) subdivision. In response to the 2 μg/kg LPS dose ( middle ), both polygonal/multipolar ( open arrows ) and round-shaped cells ( closed arrowheads ) exhibit COX-2-ir, suggesting involvement of both perivascular and endothelial cells. Fos induction under this condition is marginally increased, with greater involvement of the dorsal parvocellular ( dp ) and posterior magnocellular ( pm ) aspects of the PVH. The 100 μg/kg LPS dose ( right ) also provokes COX-2-ir expression in both polygonal/multipolar ( open arrows ) and round ( arrowheads ) cells, whose number and staining intensity are enhanced. Fos induction in the PVH is most robust under this condition and distributed uniformly throughout all subregions of the nucleus. Scale bar, 100 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Staining, Activation Assay, Expressing

    Basal and IL-1-stimulated COX-2 mRNA expression. Dark-field photomicrographs of sections from rats killed 1 hr after intravenous injection of vehicle ( top row ) or 1.87 μg/kg IL-1 ( bottom row ), at the levels of the preoptic area ( left ), paraventricular nucleus ( middle ), and medulla ( right ). In vehicle-treated rats, COX-2 mRNA is evident throughout the isocortex, hippocampal formation, and the area postrema. Some signal is also clearly evident within the meninges ( men ) and a few blood vessels ( bv ). IL-1 treatment does not appear to alter neuronal expression of COX-2 mRNA, although expression by cells associated with the vasculature and the meninges is clearly increased throughout the brain. Scale bar, 100 μm. ac , Anterior commissure; och , optic chiasm; men , meninges; OT , olfactory tubercule; CP , caudate putamen; Pir , piriform cortex; iso , isocortex; DG , dentate gyrus; CA3 , field CA3 of ammon's horn; NLOT , nucleus of lateral olfactory tract; AP , area postrema; cc , central canal; SNV , spinal nucleus of trigeminal.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Basal and IL-1-stimulated COX-2 mRNA expression. Dark-field photomicrographs of sections from rats killed 1 hr after intravenous injection of vehicle ( top row ) or 1.87 μg/kg IL-1 ( bottom row ), at the levels of the preoptic area ( left ), paraventricular nucleus ( middle ), and medulla ( right ). In vehicle-treated rats, COX-2 mRNA is evident throughout the isocortex, hippocampal formation, and the area postrema. Some signal is also clearly evident within the meninges ( men ) and a few blood vessels ( bv ). IL-1 treatment does not appear to alter neuronal expression of COX-2 mRNA, although expression by cells associated with the vasculature and the meninges is clearly increased throughout the brain. Scale bar, 100 μm. ac , Anterior commissure; och , optic chiasm; men , meninges; OT , olfactory tubercule; CP , caudate putamen; Pir , piriform cortex; iso , isocortex; DG , dentate gyrus; CA3 , field CA3 of ammon's horn; NLOT , nucleus of lateral olfactory tract; AP , area postrema; cc , central canal; SNV , spinal nucleus of trigeminal.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Expressing, Injection

    Immunoreactive COX-2 expression in the brain. Bright-field images of COX-2-ir cells in the isocortex and meninges ( top ) and cells associated with the vasculature in the forebrain ( middle ) and medulla ( bottom ), from rats killed 4 hr after vehicle ( left ) or IL-1 injection ( right ). In agreement with findings at the mRNA level, constitutive COX-2-ir is seen within some cortical neurons and, at lower levels, in the meninges and perivascular regions. At 4 hr after IL-1 (1.87 μg/kg) treatment, a clear increase in the number and staining intensity of COX-2-positive cells is seen within the meninges and perivascular regions, but not in neurons. Scale bar, 100 μm.

    Journal: The Journal of Neuroscience

    Article Title: Distinct Brain Vascular Cell Types Manifest Inducible Cyclooxygenase Expression as a Function of the Strength and Nature of Immune Insults

    doi: 10.1523/JNEUROSCI.22-13-05606.2002

    Figure Lengend Snippet: Immunoreactive COX-2 expression in the brain. Bright-field images of COX-2-ir cells in the isocortex and meninges ( top ) and cells associated with the vasculature in the forebrain ( middle ) and medulla ( bottom ), from rats killed 4 hr after vehicle ( left ) or IL-1 injection ( right ). In agreement with findings at the mRNA level, constitutive COX-2-ir is seen within some cortical neurons and, at lower levels, in the meninges and perivascular regions. At 4 hr after IL-1 (1.87 μg/kg) treatment, a clear increase in the number and staining intensity of COX-2-positive cells is seen within the meninges and perivascular regions, but not in neurons. Scale bar, 100 μm.

    Article Snippet: The material was examined in a JEOL 100 CX II transmission electron microscope.

    Techniques: Expressing, Injection, Staining