Organization of the secretory machinery in the rodent brain: Distribution of the t-SNAREs, SNAP-25 and SNAP-23

Dong Chen; Stephen L. Minger; William G. Honer; S. W. Whiteheart

doi:10.1016/S0006-8993(99)01371-2

Organization of the secretory machinery in the rodent brain: Distribution of the t-SNAREs, SNAP-25 and SNAP-23

Dong Chen, Stephen L. Minger, William G. Honer, S. W. Whiteheart

Molecular and Cellular Biochemistry

Research output: Contribution to journal › Article › peer-review

44 Scopus citations

Abstract

Vesicular transport events appear to be facilitated by the VAMP/synaptobrevin family of membrane proteins in the vesicle (v-SNAREs) and a heterodimeric complex of syntaxin and SNAP-23/25 family members in the target membrane (t-SNAREs). In this manuscript we examine the tissue distribution and composition of the heterodimeric t-SNARE complexes in adult rodent brain. Analysis of protein extracts from brain regions shows that SNAP-25, syntaxin 1, and 4 are broadly distributed, while SNAP-23, syntaxin 3, and 7 show distinct patterns of expression. Further immunohistochemistry and fractionation studies show that while SNAP-25 is enriched in axons and nerve terminals, SNAP-23 is concentrated in cell bodies. Both SNAP-23 and SNAP-25 associate with the plasma membrane and can be metabolically labeled with [³H] palmitate in AtT-20 cells. Anti-SNAP-25 antibodies co- immunoprecipitate t-SNARE heterodimers from brain extracts that predominantly contain syntaxin 1 and 2. Contrary to results from in vitro binding assays, SNAP-23 was found predominantly associated with syntaxin 3. These observations suggest that t-SNARE, heterodimer composition is governed more by SNARE expression and localization than by simple protein-protein affinity.

Original language	English
Pages (from-to)	11-24
Number of pages	14
Journal	Brain Research
Volume	831
Issue number	1-2
DOIs	https://doi.org/10.1016/S0006-8993(99)01371-2
State	Published - Jun 12 1999

Bibliographical note

Funding Information:
We thank Ramona Alcala for technical assistance and Dr. John Slevin for helpful discussions. We also thank Dr. Mark K. Bennet, Dr. Reinhard Jahn, and Dr. A.L. Hubbard for the generous gifts of their antibodies. We thank Paula Lemons, Atish Mukerjee, and Audrey Bernstein for their comments on this manuscript. This work is supported by a grant from the National Institutes of Health to S.W.W.; S.L.M. is supported by the Welcome Trust.

Funding

We thank Ramona Alcala for technical assistance and Dr. John Slevin for helpful discussions. We also thank Dr. Mark K. Bennet, Dr. Reinhard Jahn, and Dr. A.L. Hubbard for the generous gifts of their antibodies. We thank Paula Lemons, Atish Mukerjee, and Audrey Bernstein for their comments on this manuscript. This work is supported by a grant from the National Institutes of Health to S.W.W.; S.L.M. is supported by the Welcome Trust.

Funders	Funder number
National Institutes of Health (NIH)
Wellcome Trust

Keywords

Membrane trafficking
Neurons
SNAP-23
SNAP-25
Syntaxin
t-SNAREs

ASJC Scopus subject areas

General Neuroscience
Molecular Biology
Clinical Neurology
Developmental Biology

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10.1016/S0006-8993(99)01371-2

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title = "Organization of the secretory machinery in the rodent brain: Distribution of the t-SNAREs, SNAP-25 and SNAP-23",

abstract = "Vesicular transport events appear to be facilitated by the VAMP/synaptobrevin family of membrane proteins in the vesicle (v-SNAREs) and a heterodimeric complex of syntaxin and SNAP-23/25 family members in the target membrane (t-SNAREs). In this manuscript we examine the tissue distribution and composition of the heterodimeric t-SNARE complexes in adult rodent brain. Analysis of protein extracts from brain regions shows that SNAP-25, syntaxin 1, and 4 are broadly distributed, while SNAP-23, syntaxin 3, and 7 show distinct patterns of expression. Further immunohistochemistry and fractionation studies show that while SNAP-25 is enriched in axons and nerve terminals, SNAP-23 is concentrated in cell bodies. Both SNAP-23 and SNAP-25 associate with the plasma membrane and can be metabolically labeled with [3H] palmitate in AtT-20 cells. Anti-SNAP-25 antibodies co- immunoprecipitate t-SNARE heterodimers from brain extracts that predominantly contain syntaxin 1 and 2. Contrary to results from in vitro binding assays, SNAP-23 was found predominantly associated with syntaxin 3. These observations suggest that t-SNARE, heterodimer composition is governed more by SNARE expression and localization than by simple protein-protein affinity.",

keywords = "Membrane trafficking, Neurons, SNAP-23, SNAP-25, Syntaxin, t-SNAREs",

author = "Dong Chen and Minger, {Stephen L.} and Honer, {William G.} and Whiteheart, {S. W.}",

note = "Funding Information: We thank Ramona Alcala for technical assistance and Dr. John Slevin for helpful discussions. We also thank Dr. Mark K. Bennet, Dr. Reinhard Jahn, and Dr. A.L. Hubbard for the generous gifts of their antibodies. We thank Paula Lemons, Atish Mukerjee, and Audrey Bernstein for their comments on this manuscript. This work is supported by a grant from the National Institutes of Health to S.W.W.; S.L.M. is supported by the Welcome Trust.",

year = "1999",

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doi = "10.1016/S0006-8993(99)01371-2",

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AU - Honer, William G.

AU - Whiteheart, S. W.

N1 - Funding Information: We thank Ramona Alcala for technical assistance and Dr. John Slevin for helpful discussions. We also thank Dr. Mark K. Bennet, Dr. Reinhard Jahn, and Dr. A.L. Hubbard for the generous gifts of their antibodies. We thank Paula Lemons, Atish Mukerjee, and Audrey Bernstein for their comments on this manuscript. This work is supported by a grant from the National Institutes of Health to S.W.W.; S.L.M. is supported by the Welcome Trust.

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N2 - Vesicular transport events appear to be facilitated by the VAMP/synaptobrevin family of membrane proteins in the vesicle (v-SNAREs) and a heterodimeric complex of syntaxin and SNAP-23/25 family members in the target membrane (t-SNAREs). In this manuscript we examine the tissue distribution and composition of the heterodimeric t-SNARE complexes in adult rodent brain. Analysis of protein extracts from brain regions shows that SNAP-25, syntaxin 1, and 4 are broadly distributed, while SNAP-23, syntaxin 3, and 7 show distinct patterns of expression. Further immunohistochemistry and fractionation studies show that while SNAP-25 is enriched in axons and nerve terminals, SNAP-23 is concentrated in cell bodies. Both SNAP-23 and SNAP-25 associate with the plasma membrane and can be metabolically labeled with [3H] palmitate in AtT-20 cells. Anti-SNAP-25 antibodies co- immunoprecipitate t-SNARE heterodimers from brain extracts that predominantly contain syntaxin 1 and 2. Contrary to results from in vitro binding assays, SNAP-23 was found predominantly associated with syntaxin 3. These observations suggest that t-SNARE, heterodimer composition is governed more by SNARE expression and localization than by simple protein-protein affinity.

AB - Vesicular transport events appear to be facilitated by the VAMP/synaptobrevin family of membrane proteins in the vesicle (v-SNAREs) and a heterodimeric complex of syntaxin and SNAP-23/25 family members in the target membrane (t-SNAREs). In this manuscript we examine the tissue distribution and composition of the heterodimeric t-SNARE complexes in adult rodent brain. Analysis of protein extracts from brain regions shows that SNAP-25, syntaxin 1, and 4 are broadly distributed, while SNAP-23, syntaxin 3, and 7 show distinct patterns of expression. Further immunohistochemistry and fractionation studies show that while SNAP-25 is enriched in axons and nerve terminals, SNAP-23 is concentrated in cell bodies. Both SNAP-23 and SNAP-25 associate with the plasma membrane and can be metabolically labeled with [3H] palmitate in AtT-20 cells. Anti-SNAP-25 antibodies co- immunoprecipitate t-SNARE heterodimers from brain extracts that predominantly contain syntaxin 1 and 2. Contrary to results from in vitro binding assays, SNAP-23 was found predominantly associated with syntaxin 3. These observations suggest that t-SNARE, heterodimer composition is governed more by SNARE expression and localization than by simple protein-protein affinity.

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KW - Neurons

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KW - SNAP-25

KW - Syntaxin

KW - t-SNAREs

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SN - 0006-8993

VL - 831

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EP - 24

JO - Brain Research

JF - Brain Research

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ER -

Organization of the secretory machinery in the rodent brain: Distribution of the t-SNAREs, SNAP-25 and SNAP-23

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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