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Thy-1 is a component common to multiple populations of synaptic vesicles.

Jeng CJ, McCarroll SA, Martin TF, Floor E, Adams J, Krantz D, Butz S, Edwards R, Schweitzer ES - J. Cell Biol. (1998)

Bottom Line: Thy-1 is also present in synaptic vesicles in rat central nervous system.Experiments on permeabilized PC12 cells demonstrate that antibodies against Thy-1 inhibit the regulated release of neurotransmitter; this inhibition appears to be independent of any effect on the Ca2+ channel.These findings suggest Thy-1 is an integral component of many types of regulated secretory vesicles, and plays an important role in the regulated vesicular release of neurotransmitter at the synapse.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology and Brain Research Institute, UCLA School of Medicine, Los Angeles, California 90095-1763, USA.

ABSTRACT
Thy-1, a glycosylphosphatidylinositol-linked integral membrane protein of the immunoglobulin superfamily, is a component of both large dense-core and small clear vesicles in PC12 cells. A majority of this protein, formerly recognized only on the plasma membrane of neurons, is localized to regulated secretory vesicles. Thy-1 is also present in synaptic vesicles in rat central nervous system. Experiments on permeabilized PC12 cells demonstrate that antibodies against Thy-1 inhibit the regulated release of neurotransmitter; this inhibition appears to be independent of any effect on the Ca2+ channel. These findings suggest Thy-1 is an integral component of many types of regulated secretory vesicles, and plays an important role in the regulated vesicular release of neurotransmitter at the synapse.

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Localization of Thy-1 immunoreactivity in rat brain. (A–C) Localization of 7C8  immunoreactivity in the hippocampus. Sections of rat hippocampus were stained for  Thy-1 (A), SV2 (B), or synaptophysin (brown  HRP reaction product), and counterstained  with methylene green. In coronal sections of  the hippocampus, 7C8 immunoreactivity was  found throughout the synaptic fields, including the stratum oriens, stratum radiatum, and  the molecular layer of the dentate gyrus. In  contrast, there was nothing labeled with the  antibody in the pyramidal cell layer, stratum  lacunosum moleculare and granular layer of  the dentate gyrus. Both SV2 and synaptophysin immunoreactivities were similar to  that of Thy-1, and were distributed throughout the synaptic fields, but not in the cell  body layers. (D–F) Thy-1 copurifies with  large and small synaptic vesicles. The distribution of Thy-1 (D), SV2 (E), and synaptophysin (F) immunoreactivities were assayed  by dot blot after fractionation of membranes  by Sephacryl S-1000 column chromatography.  The three arrows indicate the positions of the  void volume of the column (V0), the elution  position of large, dense-core vesicles (LDV),  and the elution position of the small synaptic  vesicles (SSV). Data shown are the mean ±  range of duplicated determinations. (G–I)  Localization of 7C8 immunoreactivity in the  retina. Sections of rat retina were stained for  immunoreactivity to 7C8 and then counterstained with methylene green. The sections  are oriented with the photoreceptor layer at  the bottom. 7C8 immunoreactivity was prominently in the ganglion cell layer (gc) and  faintly in the inner plexiform layer (ip). In  contrast to SV2 (H), and synaptophysin (I),  7C8 immunoreactivity was largely absent  from the inner nuclear layer (in), outer plexiform layer (op), and outer nuclear layer (on).  Bars: (A–C) 500 μm; (G–I) 100 μm.
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Figure 9: Localization of Thy-1 immunoreactivity in rat brain. (A–C) Localization of 7C8 immunoreactivity in the hippocampus. Sections of rat hippocampus were stained for Thy-1 (A), SV2 (B), or synaptophysin (brown HRP reaction product), and counterstained with methylene green. In coronal sections of the hippocampus, 7C8 immunoreactivity was found throughout the synaptic fields, including the stratum oriens, stratum radiatum, and the molecular layer of the dentate gyrus. In contrast, there was nothing labeled with the antibody in the pyramidal cell layer, stratum lacunosum moleculare and granular layer of the dentate gyrus. Both SV2 and synaptophysin immunoreactivities were similar to that of Thy-1, and were distributed throughout the synaptic fields, but not in the cell body layers. (D–F) Thy-1 copurifies with large and small synaptic vesicles. The distribution of Thy-1 (D), SV2 (E), and synaptophysin (F) immunoreactivities were assayed by dot blot after fractionation of membranes by Sephacryl S-1000 column chromatography. The three arrows indicate the positions of the void volume of the column (V0), the elution position of large, dense-core vesicles (LDV), and the elution position of the small synaptic vesicles (SSV). Data shown are the mean ± range of duplicated determinations. (G–I) Localization of 7C8 immunoreactivity in the retina. Sections of rat retina were stained for immunoreactivity to 7C8 and then counterstained with methylene green. The sections are oriented with the photoreceptor layer at the bottom. 7C8 immunoreactivity was prominently in the ganglion cell layer (gc) and faintly in the inner plexiform layer (ip). In contrast to SV2 (H), and synaptophysin (I), 7C8 immunoreactivity was largely absent from the inner nuclear layer (in), outer plexiform layer (op), and outer nuclear layer (on). Bars: (A–C) 500 μm; (G–I) 100 μm.

Mentions: To demonstrate that the vesicular localization of Thy-1 is not unique to the PC12 cell line, we analyzed rat brain tissue for the presence and localization of Thy-1. Western blot analysis of various brain regions and other tissues demonstrated that 7C8 bound to virtually all synapse-containing regions of the central nervous system, but was undetectable in peripheral tissues, including liver, kidney, pancreas, and muscle (data not shown). Extensive immunohistochemical mapping of the distribution of 7C8 binding throughout the nervous system showed that 7C8 binding was prominent in the gray matter in synapse-rich areas, and that this staining paralleled that seen with other vesicle markers. One example of this general colocalization is shown in Fig. 9 A, in which the synaptic layers of the hippocampus stain prominently with 7C8 (A), SV2 (B), and synaptophysin (C). Although we can identify subtle differences in the relative staining of vesicles markers in some regions (for example, in the dentate gyrus of the hippocampus, there is relatively less SV2 staining in the vicinity of the granule cell layer as compared with 7C8 or synaptophysin); most areas show similar staining of all these vesicle markers. This result suggests that Thy-1 is somehow associated with synapses.


Thy-1 is a component common to multiple populations of synaptic vesicles.

Jeng CJ, McCarroll SA, Martin TF, Floor E, Adams J, Krantz D, Butz S, Edwards R, Schweitzer ES - J. Cell Biol. (1998)

Localization of Thy-1 immunoreactivity in rat brain. (A–C) Localization of 7C8  immunoreactivity in the hippocampus. Sections of rat hippocampus were stained for  Thy-1 (A), SV2 (B), or synaptophysin (brown  HRP reaction product), and counterstained  with methylene green. In coronal sections of  the hippocampus, 7C8 immunoreactivity was  found throughout the synaptic fields, including the stratum oriens, stratum radiatum, and  the molecular layer of the dentate gyrus. In  contrast, there was nothing labeled with the  antibody in the pyramidal cell layer, stratum  lacunosum moleculare and granular layer of  the dentate gyrus. Both SV2 and synaptophysin immunoreactivities were similar to  that of Thy-1, and were distributed throughout the synaptic fields, but not in the cell  body layers. (D–F) Thy-1 copurifies with  large and small synaptic vesicles. The distribution of Thy-1 (D), SV2 (E), and synaptophysin (F) immunoreactivities were assayed  by dot blot after fractionation of membranes  by Sephacryl S-1000 column chromatography.  The three arrows indicate the positions of the  void volume of the column (V0), the elution  position of large, dense-core vesicles (LDV),  and the elution position of the small synaptic  vesicles (SSV). Data shown are the mean ±  range of duplicated determinations. (G–I)  Localization of 7C8 immunoreactivity in the  retina. Sections of rat retina were stained for  immunoreactivity to 7C8 and then counterstained with methylene green. The sections  are oriented with the photoreceptor layer at  the bottom. 7C8 immunoreactivity was prominently in the ganglion cell layer (gc) and  faintly in the inner plexiform layer (ip). In  contrast to SV2 (H), and synaptophysin (I),  7C8 immunoreactivity was largely absent  from the inner nuclear layer (in), outer plexiform layer (op), and outer nuclear layer (on).  Bars: (A–C) 500 μm; (G–I) 100 μm.
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Related In: Results  -  Collection

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Figure 9: Localization of Thy-1 immunoreactivity in rat brain. (A–C) Localization of 7C8 immunoreactivity in the hippocampus. Sections of rat hippocampus were stained for Thy-1 (A), SV2 (B), or synaptophysin (brown HRP reaction product), and counterstained with methylene green. In coronal sections of the hippocampus, 7C8 immunoreactivity was found throughout the synaptic fields, including the stratum oriens, stratum radiatum, and the molecular layer of the dentate gyrus. In contrast, there was nothing labeled with the antibody in the pyramidal cell layer, stratum lacunosum moleculare and granular layer of the dentate gyrus. Both SV2 and synaptophysin immunoreactivities were similar to that of Thy-1, and were distributed throughout the synaptic fields, but not in the cell body layers. (D–F) Thy-1 copurifies with large and small synaptic vesicles. The distribution of Thy-1 (D), SV2 (E), and synaptophysin (F) immunoreactivities were assayed by dot blot after fractionation of membranes by Sephacryl S-1000 column chromatography. The three arrows indicate the positions of the void volume of the column (V0), the elution position of large, dense-core vesicles (LDV), and the elution position of the small synaptic vesicles (SSV). Data shown are the mean ± range of duplicated determinations. (G–I) Localization of 7C8 immunoreactivity in the retina. Sections of rat retina were stained for immunoreactivity to 7C8 and then counterstained with methylene green. The sections are oriented with the photoreceptor layer at the bottom. 7C8 immunoreactivity was prominently in the ganglion cell layer (gc) and faintly in the inner plexiform layer (ip). In contrast to SV2 (H), and synaptophysin (I), 7C8 immunoreactivity was largely absent from the inner nuclear layer (in), outer plexiform layer (op), and outer nuclear layer (on). Bars: (A–C) 500 μm; (G–I) 100 μm.
Mentions: To demonstrate that the vesicular localization of Thy-1 is not unique to the PC12 cell line, we analyzed rat brain tissue for the presence and localization of Thy-1. Western blot analysis of various brain regions and other tissues demonstrated that 7C8 bound to virtually all synapse-containing regions of the central nervous system, but was undetectable in peripheral tissues, including liver, kidney, pancreas, and muscle (data not shown). Extensive immunohistochemical mapping of the distribution of 7C8 binding throughout the nervous system showed that 7C8 binding was prominent in the gray matter in synapse-rich areas, and that this staining paralleled that seen with other vesicle markers. One example of this general colocalization is shown in Fig. 9 A, in which the synaptic layers of the hippocampus stain prominently with 7C8 (A), SV2 (B), and synaptophysin (C). Although we can identify subtle differences in the relative staining of vesicles markers in some regions (for example, in the dentate gyrus of the hippocampus, there is relatively less SV2 staining in the vicinity of the granule cell layer as compared with 7C8 or synaptophysin); most areas show similar staining of all these vesicle markers. This result suggests that Thy-1 is somehow associated with synapses.

Bottom Line: Thy-1 is also present in synaptic vesicles in rat central nervous system.Experiments on permeabilized PC12 cells demonstrate that antibodies against Thy-1 inhibit the regulated release of neurotransmitter; this inhibition appears to be independent of any effect on the Ca2+ channel.These findings suggest Thy-1 is an integral component of many types of regulated secretory vesicles, and plays an important role in the regulated vesicular release of neurotransmitter at the synapse.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurobiology and Brain Research Institute, UCLA School of Medicine, Los Angeles, California 90095-1763, USA.

ABSTRACT
Thy-1, a glycosylphosphatidylinositol-linked integral membrane protein of the immunoglobulin superfamily, is a component of both large dense-core and small clear vesicles in PC12 cells. A majority of this protein, formerly recognized only on the plasma membrane of neurons, is localized to regulated secretory vesicles. Thy-1 is also present in synaptic vesicles in rat central nervous system. Experiments on permeabilized PC12 cells demonstrate that antibodies against Thy-1 inhibit the regulated release of neurotransmitter; this inhibition appears to be independent of any effect on the Ca2+ channel. These findings suggest Thy-1 is an integral component of many types of regulated secretory vesicles, and plays an important role in the regulated vesicular release of neurotransmitter at the synapse.

Show MeSH
Related in: MedlinePlus