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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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Related in: MedlinePlus

Expression of the  7C8 antigen in COS cells. (A)  Homogenates of COS cells  transfected with two of the  pcDM8 expression plasmids  isolated from immunoreactive CHO cell clones were  stained on a Western blot with the 7C8 antibody. Clone 112 expressed a protein that comigrated with the 7C8 antigen expressed  by PC12 cells, but untransfected COS cells (−) and cells transfected with plasmid 11 did not. (B) The cDNA insert from plasmid 112 was subcloned into the expression vector pcDNA I-Amp.  Homogenates from COS cells transfected with the cDNA insert  in the sense (S) but not the antisense (A) of the Thy-1 cDNA reacted with the 7C8 antibody.
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Figure 4: Expression of the 7C8 antigen in COS cells. (A) Homogenates of COS cells transfected with two of the pcDM8 expression plasmids isolated from immunoreactive CHO cell clones were stained on a Western blot with the 7C8 antibody. Clone 112 expressed a protein that comigrated with the 7C8 antigen expressed by PC12 cells, but untransfected COS cells (−) and cells transfected with plasmid 11 did not. (B) The cDNA insert from plasmid 112 was subcloned into the expression vector pcDNA I-Amp. Homogenates from COS cells transfected with the cDNA insert in the sense (S) but not the antisense (A) of the Thy-1 cDNA reacted with the 7C8 antibody.

Mentions: To determine which of these two plasmids encoded the 7C8 antigen, we transfected each plasmid separately into COS cells, and probed Western blots of the cell homogenates with the 7C8 antibody. Only plasmid 112 showed an immunoreactive protein of the expected size (Fig. 4 A). Analysis of the 1.6-kb insert showed sequence identity to rat Thy-1 (Moriuchi et al., 1983; Seki et al., 1985) beginning 16 nucleotides upstream of the initiation codon and extending through the entire protein-coding regions as well as an additional 1,053 base pairs of 3′ untranslated DNA. Thy-1 is a GPI-linked protein known to be on the surface of T lymphocytes and mature neurons, and has an apparent molecular weight of ∼27 kD; it has been reported to run occasionally as a doublet due to heterogeneity in its N-linked glycosylation (Barboni et al., 1995).


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)

Expression of the  7C8 antigen in COS cells. (A)  Homogenates of COS cells  transfected with two of the  pcDM8 expression plasmids  isolated from immunoreactive CHO cell clones were  stained on a Western blot with the 7C8 antibody. Clone 112 expressed a protein that comigrated with the 7C8 antigen expressed  by PC12 cells, but untransfected COS cells (−) and cells transfected with plasmid 11 did not. (B) The cDNA insert from plasmid 112 was subcloned into the expression vector pcDNA I-Amp.  Homogenates from COS cells transfected with the cDNA insert  in the sense (S) but not the antisense (A) of the Thy-1 cDNA reacted with the 7C8 antibody.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2140167&req=5

Figure 4: Expression of the 7C8 antigen in COS cells. (A) Homogenates of COS cells transfected with two of the pcDM8 expression plasmids isolated from immunoreactive CHO cell clones were stained on a Western blot with the 7C8 antibody. Clone 112 expressed a protein that comigrated with the 7C8 antigen expressed by PC12 cells, but untransfected COS cells (−) and cells transfected with plasmid 11 did not. (B) The cDNA insert from plasmid 112 was subcloned into the expression vector pcDNA I-Amp. Homogenates from COS cells transfected with the cDNA insert in the sense (S) but not the antisense (A) of the Thy-1 cDNA reacted with the 7C8 antibody.
Mentions: To determine which of these two plasmids encoded the 7C8 antigen, we transfected each plasmid separately into COS cells, and probed Western blots of the cell homogenates with the 7C8 antibody. Only plasmid 112 showed an immunoreactive protein of the expected size (Fig. 4 A). Analysis of the 1.6-kb insert showed sequence identity to rat Thy-1 (Moriuchi et al., 1983; Seki et al., 1985) beginning 16 nucleotides upstream of the initiation codon and extending through the entire protein-coding regions as well as an additional 1,053 base pairs of 3′ untranslated DNA. Thy-1 is a GPI-linked protein known to be on the surface of T lymphocytes and mature neurons, and has an apparent molecular weight of ∼27 kD; it has been reported to run occasionally as a doublet due to heterogeneity in its N-linked glycosylation (Barboni et al., 1995).

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