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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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Identification of 7C8 antigen as Thy-1. (A) OX7 immunoprecipitation. Postnuclear supernatant of PC12 cells (PC12 S1)  was solubilized with Det-HBS, and immunoprecipitated with OX7  coated-SAC (OX7) or with normal mouse serum coated-SAC  (NMS). Supernatant (S) and pellet (P) fractions after low speed  centrifugation were subjected to a nonreducing SDS-PAGE and  immunoblotting with OX7 (lanes 1–5) or 7C8 (lanes 6–10) antibodies. (B) Immunoblots of OX7 and 7C8. Homogenates (100 μg  total protein per lane) of PC12 cells, rat brain, BALB/c mouse  brain, and AKR mouse brain were loaded on a nonreducing gel,  transferred to nitrocellulose membranes, and probed with OX7  (lanes 1–4) or 7C8 (lanes 5–8) antibodies.
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Figure 5: Identification of 7C8 antigen as Thy-1. (A) OX7 immunoprecipitation. Postnuclear supernatant of PC12 cells (PC12 S1) was solubilized with Det-HBS, and immunoprecipitated with OX7 coated-SAC (OX7) or with normal mouse serum coated-SAC (NMS). Supernatant (S) and pellet (P) fractions after low speed centrifugation were subjected to a nonreducing SDS-PAGE and immunoblotting with OX7 (lanes 1–5) or 7C8 (lanes 6–10) antibodies. (B) Immunoblots of OX7 and 7C8. Homogenates (100 μg total protein per lane) of PC12 cells, rat brain, BALB/c mouse brain, and AKR mouse brain were loaded on a nonreducing gel, transferred to nitrocellulose membranes, and probed with OX7 (lanes 1–4) or 7C8 (lanes 5–8) antibodies.

Mentions: Biochemical experiments confirmed that the 7C8 antigen is Thy-1. Fig. 5 A shows that the 7C8 antibody binds to the same protein recognized by a commercially available monoclonal antibody against Thy-1 (OX7). We immunoprecipitated Thy-1 from a detergent extract of a PC12 cell postnuclear supernatant. As shown in the first panel, illustrating an immunoblot probed with the same OX7 anti– Thy-1 antibody used for the immunoprecipitation, all of the Thy-1 in the homogenate was pelleted in the immunoprecipitation; in contrast, normal mouse serum leaves all the Thy-1 in the supernatant. The second panel in Fig. 5 A illustrates that probing these samples with the 7C8 antibody gives exactly the same pattern as for OX7; i.e., the 7C8 antigen is precipitated by the OX7 antibody, but not by control serum. A comparison of the two panels in Fig. 5 A also illustrates the identical mobility of the proteins recognized by these two antibodies.


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)

Identification of 7C8 antigen as Thy-1. (A) OX7 immunoprecipitation. Postnuclear supernatant of PC12 cells (PC12 S1)  was solubilized with Det-HBS, and immunoprecipitated with OX7  coated-SAC (OX7) or with normal mouse serum coated-SAC  (NMS). Supernatant (S) and pellet (P) fractions after low speed  centrifugation were subjected to a nonreducing SDS-PAGE and  immunoblotting with OX7 (lanes 1–5) or 7C8 (lanes 6–10) antibodies. (B) Immunoblots of OX7 and 7C8. Homogenates (100 μg  total protein per lane) of PC12 cells, rat brain, BALB/c mouse  brain, and AKR mouse brain were loaded on a nonreducing gel,  transferred to nitrocellulose membranes, and probed with OX7  (lanes 1–4) or 7C8 (lanes 5–8) antibodies.
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Related In: Results  -  Collection

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Figure 5: Identification of 7C8 antigen as Thy-1. (A) OX7 immunoprecipitation. Postnuclear supernatant of PC12 cells (PC12 S1) was solubilized with Det-HBS, and immunoprecipitated with OX7 coated-SAC (OX7) or with normal mouse serum coated-SAC (NMS). Supernatant (S) and pellet (P) fractions after low speed centrifugation were subjected to a nonreducing SDS-PAGE and immunoblotting with OX7 (lanes 1–5) or 7C8 (lanes 6–10) antibodies. (B) Immunoblots of OX7 and 7C8. Homogenates (100 μg total protein per lane) of PC12 cells, rat brain, BALB/c mouse brain, and AKR mouse brain were loaded on a nonreducing gel, transferred to nitrocellulose membranes, and probed with OX7 (lanes 1–4) or 7C8 (lanes 5–8) antibodies.
Mentions: Biochemical experiments confirmed that the 7C8 antigen is Thy-1. Fig. 5 A shows that the 7C8 antibody binds to the same protein recognized by a commercially available monoclonal antibody against Thy-1 (OX7). We immunoprecipitated Thy-1 from a detergent extract of a PC12 cell postnuclear supernatant. As shown in the first panel, illustrating an immunoblot probed with the same OX7 anti– Thy-1 antibody used for the immunoprecipitation, all of the Thy-1 in the homogenate was pelleted in the immunoprecipitation; in contrast, normal mouse serum leaves all the Thy-1 in the supernatant. The second panel in Fig. 5 A illustrates that probing these samples with the 7C8 antibody gives exactly the same pattern as for OX7; i.e., the 7C8 antigen is precipitated by the OX7 antibody, but not by control serum. A comparison of the two panels in Fig. 5 A also illustrates the identical mobility of the proteins recognized by these two antibodies.

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