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Expression and immunolocalization of Gpnmb, a glioma-associated glycoprotein, in normal and inflamed central nervous systems of adult rats.

Huang JJ, Ma WJ, Yokoyama S - Brain Behav (2012)

Bottom Line: Double immunofluorescence staining showed that these cells were co-stained most frequently with the microglia/macrophage marker OX42, and occasionally with the radial glia marker RC2 or the neuronal marker NeuN.Furthermore, an intraperitoneal injection of bacterial endotoxin lipopolysaccharide increased the number of Gpnmb and OX42 double-positive cells in the area postrema, which is one of the circumventricular organs, indicating infiltration of hematogenous macrophages.These results suggest that Gpnmb, which is expressed in microglia and macrophages in non-tumorous neural tissues, plays an important role in the regulation of immune/inflammatory responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysical Genetics, Kanazawa University Graduate School of Medicine Kanazawa 920-8640, Japan.

ABSTRACT
Glycoprotein nonmetastatic melanoma B (Gpnmb) is a type I transmembrane protein implicated in cell differentiation, inflammation, tissue regeneration, and tumor progression. Gpnmb, which is highly expressed in glioblastoma cells, is a potential therapeutic target. However, little is known about its expression, cellular localization, and roles in non-tumorous neural tissues. In this study, we examined Gpnmb expression in the central nervous system of adult rats under both normal and inflammatory conditions. Reverse transcription-polymerase chain reaction analysis revealed that Gpnmb mRNA was expressed in the cerebrum, cerebellum, brain stem, and spinal cord of normal adult rats. Immunoperoxidase staining revealed that Gpnmb-immunoreactive cells were widely distributed in the parenchyma of all brain regions examined, with the cells being most prevalent in the hippocampal dentate gyrus, cerebellar cortex, spinal dorsal horn, choroid plexus, ependyma, periventricular regions, and in layers II and III of the cerebral cortex. Double immunofluorescence staining showed that these cells were co-stained most frequently with the microglia/macrophage marker OX42, and occasionally with the radial glia marker RC2 or the neuronal marker NeuN. Furthermore, an intraperitoneal injection of bacterial endotoxin lipopolysaccharide increased the number of Gpnmb and OX42 double-positive cells in the area postrema, which is one of the circumventricular organs, indicating infiltration of hematogenous macrophages. These results suggest that Gpnmb, which is expressed in microglia and macrophages in non-tumorous neural tissues, plays an important role in the regulation of immune/inflammatory responses.

No MeSH data available.


Related in: MedlinePlus

Gpnmb-IR in rat cerebellum. (A–C) Immunoperoxidase staining of rat cerebellar cortex. (A, B) Low magnification images. Sections obtained from adult rats were stained with the anti-Gpnmb antibody before [ A, ads (-)] or after [ B, ads (+)] adsorption with the antigenic peptide and then visualized with DAB. (C) Gpnmb-IR in cell bodies in the Purkinje cell layer and processes in the molecular layer are indicated by arrows and arrowheads, respectively. (D–F) Double immunofluorescence staining of rat cerebellar cortex. Sections from adult rats were double-stained for Gpnmb (green) and the indicated markers (red). Note that Gpnmb-IR is co-localized with GFAP (D) and RC2 (E) but not with calbindin D-28K (F). ML, molecular layer; PCL, Purkinje cell layer; GCL, granule cell layer. Scale bars: A, B, 30 μm; C–F, 20 μm.
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fig07: Gpnmb-IR in rat cerebellum. (A–C) Immunoperoxidase staining of rat cerebellar cortex. (A, B) Low magnification images. Sections obtained from adult rats were stained with the anti-Gpnmb antibody before [ A, ads (-)] or after [ B, ads (+)] adsorption with the antigenic peptide and then visualized with DAB. (C) Gpnmb-IR in cell bodies in the Purkinje cell layer and processes in the molecular layer are indicated by arrows and arrowheads, respectively. (D–F) Double immunofluorescence staining of rat cerebellar cortex. Sections from adult rats were double-stained for Gpnmb (green) and the indicated markers (red). Note that Gpnmb-IR is co-localized with GFAP (D) and RC2 (E) but not with calbindin D-28K (F). ML, molecular layer; PCL, Purkinje cell layer; GCL, granule cell layer. Scale bars: A, B, 30 μm; C–F, 20 μm.

Mentions: In the cerebellum, cell bodies in the Purkinje cell layer and fine processes in the molecular layer were stained (Fig. 7A, C). Staining was abolished by preadsorbing the primary antibody with the peptide used for immunaization (Fig. 7B). The Gpnmb-IR cells were co-stained with anti-GFAP (Fig. 7D) and anti-RC2 (Fig. 7E) antibodies, but not at all with an antibody against calbindin D-28K, a specific marker for Purkinje neurons (Fig. 7F). Therefore, we concluded that Gpnmb-IR cells were Bergmann glial cells.


Expression and immunolocalization of Gpnmb, a glioma-associated glycoprotein, in normal and inflamed central nervous systems of adult rats.

Huang JJ, Ma WJ, Yokoyama S - Brain Behav (2012)

Gpnmb-IR in rat cerebellum. (A–C) Immunoperoxidase staining of rat cerebellar cortex. (A, B) Low magnification images. Sections obtained from adult rats were stained with the anti-Gpnmb antibody before [ A, ads (-)] or after [ B, ads (+)] adsorption with the antigenic peptide and then visualized with DAB. (C) Gpnmb-IR in cell bodies in the Purkinje cell layer and processes in the molecular layer are indicated by arrows and arrowheads, respectively. (D–F) Double immunofluorescence staining of rat cerebellar cortex. Sections from adult rats were double-stained for Gpnmb (green) and the indicated markers (red). Note that Gpnmb-IR is co-localized with GFAP (D) and RC2 (E) but not with calbindin D-28K (F). ML, molecular layer; PCL, Purkinje cell layer; GCL, granule cell layer. Scale bars: A, B, 30 μm; C–F, 20 μm.
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Related In: Results  -  Collection

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fig07: Gpnmb-IR in rat cerebellum. (A–C) Immunoperoxidase staining of rat cerebellar cortex. (A, B) Low magnification images. Sections obtained from adult rats were stained with the anti-Gpnmb antibody before [ A, ads (-)] or after [ B, ads (+)] adsorption with the antigenic peptide and then visualized with DAB. (C) Gpnmb-IR in cell bodies in the Purkinje cell layer and processes in the molecular layer are indicated by arrows and arrowheads, respectively. (D–F) Double immunofluorescence staining of rat cerebellar cortex. Sections from adult rats were double-stained for Gpnmb (green) and the indicated markers (red). Note that Gpnmb-IR is co-localized with GFAP (D) and RC2 (E) but not with calbindin D-28K (F). ML, molecular layer; PCL, Purkinje cell layer; GCL, granule cell layer. Scale bars: A, B, 30 μm; C–F, 20 μm.
Mentions: In the cerebellum, cell bodies in the Purkinje cell layer and fine processes in the molecular layer were stained (Fig. 7A, C). Staining was abolished by preadsorbing the primary antibody with the peptide used for immunaization (Fig. 7B). The Gpnmb-IR cells were co-stained with anti-GFAP (Fig. 7D) and anti-RC2 (Fig. 7E) antibodies, but not at all with an antibody against calbindin D-28K, a specific marker for Purkinje neurons (Fig. 7F). Therefore, we concluded that Gpnmb-IR cells were Bergmann glial cells.

Bottom Line: Double immunofluorescence staining showed that these cells were co-stained most frequently with the microglia/macrophage marker OX42, and occasionally with the radial glia marker RC2 or the neuronal marker NeuN.Furthermore, an intraperitoneal injection of bacterial endotoxin lipopolysaccharide increased the number of Gpnmb and OX42 double-positive cells in the area postrema, which is one of the circumventricular organs, indicating infiltration of hematogenous macrophages.These results suggest that Gpnmb, which is expressed in microglia and macrophages in non-tumorous neural tissues, plays an important role in the regulation of immune/inflammatory responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysical Genetics, Kanazawa University Graduate School of Medicine Kanazawa 920-8640, Japan.

ABSTRACT
Glycoprotein nonmetastatic melanoma B (Gpnmb) is a type I transmembrane protein implicated in cell differentiation, inflammation, tissue regeneration, and tumor progression. Gpnmb, which is highly expressed in glioblastoma cells, is a potential therapeutic target. However, little is known about its expression, cellular localization, and roles in non-tumorous neural tissues. In this study, we examined Gpnmb expression in the central nervous system of adult rats under both normal and inflammatory conditions. Reverse transcription-polymerase chain reaction analysis revealed that Gpnmb mRNA was expressed in the cerebrum, cerebellum, brain stem, and spinal cord of normal adult rats. Immunoperoxidase staining revealed that Gpnmb-immunoreactive cells were widely distributed in the parenchyma of all brain regions examined, with the cells being most prevalent in the hippocampal dentate gyrus, cerebellar cortex, spinal dorsal horn, choroid plexus, ependyma, periventricular regions, and in layers II and III of the cerebral cortex. Double immunofluorescence staining showed that these cells were co-stained most frequently with the microglia/macrophage marker OX42, and occasionally with the radial glia marker RC2 or the neuronal marker NeuN. Furthermore, an intraperitoneal injection of bacterial endotoxin lipopolysaccharide increased the number of Gpnmb and OX42 double-positive cells in the area postrema, which is one of the circumventricular organs, indicating infiltration of hematogenous macrophages. These results suggest that Gpnmb, which is expressed in microglia and macrophages in non-tumorous neural tissues, plays an important role in the regulation of immune/inflammatory responses.

No MeSH data available.


Related in: MedlinePlus