Microglial Cells in the Central Nervous System of the Rabbit and Rat: Cytochemical Identification Using Two Different Lectins

Boya, J; Carbonell, A.L.; Calvo, J.; Borregón, A.

doi:10.1159/000147064

Cited by 31 publications

(20 citation statements)

References 12 publications

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“…2A). These cells displayed a ramified morphology characteristic of resting microglia in mouse, rat, rabbit, human and cat retinae described by earlier workers (Boya et al, 1991;Chan-Ling, 1994;Perry et al, 1985;Schnitzer, 1989;Penfold et al;1993). In contrast to the NDPase-labeled resting microglia, varicosities were not prominent on their processes.…”

Section: Griffonia Simplicifolia Lectin-b4 Histochemistrymentioning

confidence: 49%

Early activation of microglia in the pathogenesis of fatal murine cerebral malaria

Medana

Hunt

Chan‐Ling

1997

Glia

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Section: Griffonia Simplicifolia Lectin-b4 Histochemistrymentioning

confidence: 49%

Early activation of microglia in the pathogenesis of fatal murine cerebral malaria

Medana

Hunt

Chan‐Ling

1997

Glia

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“…However, astrocytes were negative for both markers (data not shown). Lectin receptors which are widely accepted as another marker for microglia [28][29][30] have been used to verify astrocyte-supported microglia. RCA 120, which recognizes ß-D-galactose (associated with glycoproteins on the membrane of microglia), is present on over 79% of astrocyte-supported microglia ( fig.…”

Section: Resultsmentioning

confidence: 99%

IL-1-Alpha and TNF-Alpha Differentially Regulate CD4 and Mac-1 Expression in Mouse Microglia

Zhang²,

Magistretti³

et al. 1998

Neuroimmunomodulation

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The regulatory effects of the proinflammatory cytokines, interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α) were investigated on CD4 and Mac-1 expression in mouse microglial cultures. The identity of the microglia in cultures was confirmed by multiple indices including morphology, uptake of acetylated low-density lipoprotein and lectin RCA 120 staining. Microglia growing on a monolayer of astrocytes (astrocyte-supported microglia) were both CD4- and Mac-1 positive (out of 94.5% Mac-1-positive cells, 85.3% were also CD4 positive). When astrocyte-supported microglia were replated directly onto culture dishes (plate-supported microglia), the percentage of CD4- and Mac-1-positive cells decreased to 12–29 and 20–25% resepctively. The addition of IL-1α or TNF-α to plate-supported microglia led to an upregulation of Mac-1 expression in a time- and dose-dependent manner with different EC50s (0.5 ng/ml for IL-1α and 2 ng/ml for TNF-α) but exhibited similar time-to-peak responses (over 12 h). The addition of IL-1α, but not TNF-α, also led to an increase in CD4 expression on plate-supported microglia with a similar dose response and time course. IL-1α treatment gave rise to an increase in the level of CD4 mRNA as assessed by RT-PCR. The possibility that cell proliferation was responsible for the observed effects on microglia was excluded by an analysis of ³H-thymidine incorporation. Our results suggest that cultured mouse microglia express CD4 molecules which can be upregulated by IL-1α while Mac-1 can be upregulated by both IL-1α and TNF-α.

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“…HAPI cells possess the morphology of macrophages, are capable of phagocytosis and label with isolectin B4. B4 is derived from Griffinia simplicifolia and recognizes ␣-D-galactose residues commonly found in microglia/macrophages; it is used as a selective marker for microglia (Streit and Kreutzberg, 1987;Streit, 1990;Boya et al, 1991;Kaur and Ling, 1991;Colton et al, 1992, Cheepsunthorn et al, 1998). We further demonstrate that HAPI cells, like microglia and brain macrophages, express complement 3 receptors, which are recognized by monoclonal antibody OX-42 (Milligan et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

Characterization of a novel brain‐derived microglial cell line isolated from neonatal rat brain

Cheepsunthorn

Radov

Menzies

et al. 2001

Glia

148

114

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We observed highly aggressively proliferating immortalized (HAPI) cells growing in cultures that had been enriched for microglia. The cells were initially obtained from mixed glial cultures prepared from 3-day-old rat brains. HAPI cells are typically round with few or no processes when cultured in 10% serum containing medium. As the percentage of serum in the medium is decreased, the HAPI cells have more processes. HAPI cells stain for the isolectin B4, OX-42, and GLUT5, which are markers for microglial cells, but the cells do not immunolabel with A2B5, a marker of cells in the oligodendroglial cell lineage, or with the astrocyte-specific marker, glial fibrillary aciidic protein (GFAP). In addition, HAPI cells are capable of phagocytosis. We conclude that HAPI cells are of microglia/macrophage lineage. Exposing HAPI cells to lipopolysaccharide (LPS) induces the mRNAs for tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS). LPS exposure also induces secretion of TNF-alpha and production of nitric oxide (NO) in HAPI cells. Because activation of microglia is associated with an increase in iron accumulation and ferritin expression, we tested the hypothesis that iron status affects the production of TNF-alpha and NO. Our studies demonstrate that both iron chelation and iron loading diminished the LPS-induced effect of TNF-alpha and NO. The results of this study indicate that HAPI cells possess the characteristics of microglia/brain macrophages, providing an alternative cell culture model for the study of microglia. In addition, we demonstrate that the activation of microglial cells could be modified by iron.

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Microglial Cells in the Central Nervous System of the Rabbit and Rat: Cytochemical Identification Using Two Different Lectins

Cited by 31 publications

References 12 publications

Early activation of microglia in the pathogenesis of fatal murine cerebral malaria

Early activation of microglia in the pathogenesis of fatal murine cerebral malaria

IL-1-Alpha and TNF-Alpha Differentially Regulate CD4 and Mac-1 Expression in Mouse Microglia

Characterization of a novel brain‐derived microglial cell line isolated from neonatal rat brain

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