Regulation of Podosome Formation in Macrophages by a Splice Variant of the Sodium Channel SCN8A
Voltage-gated sodium channels initiate electrical signaling in excitable cells such as muscle and neurons. They also are expressed in non-excitable cells such as macrophages and neoplastic cells. Previously, in macrophages, we demonstrated expression of SCN8A, the gene that encodes the channel NaV1....
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Veröffentlicht in: | The Journal of biological chemistry 2009-03, Vol.284 (12), p.8114-8126 |
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creator | Carrithers, Michael D. Chatterjee, Gouri Carrithers, Lisette M. Offoha, Roosevelt Iheagwara, Uzoma Rahner, Christoph Graham, Morven Waxman, Stephen G. |
description | Voltage-gated sodium channels initiate electrical signaling in excitable cells such as muscle and neurons. They also are expressed in non-excitable cells such as macrophages and neoplastic cells. Previously, in macrophages, we demonstrated expression of SCN8A, the gene that encodes the channel NaV1.6, and intracellular localization of NaV1.6 to regions near F-actin bundles, particularly at areas of cell attachment. Here we show that a splice variant of NaV1.6 regulates cellular invasion through its effects on podosome and invadopodia formation in macrophages and melanoma cells. cDNA sequence analysis of SCN8A from THP-1 cells, a human monocyte-macrophage cell line, confirmed the expression of a full-length splice variant that lacks exon 18. Immunoelectron microscopy demonstrated NaV1.6-positive staining within the electron dense podosome rosette structure. Pharmacologic antagonism with tetrodotoxin (TTX) in differentiated THP-1 cells or absence of functional NaV1.6 through a naturally occurring mutation (med) in mouse peritoneal macrophages inhibited podosome formation. Agonist-mediated activation of the channel with veratridine caused release of sodium from cationic vesicular compartments, uptake by mitochondria, and mitochondrial calcium release through the Na/Ca exchanger. Invasion by differentiated THP-1 and HTB-66 cells, an invasive melanoma cell line, through extracellular matrix was inhibited by TTX. THP-1 invasion also was inhibited by small hairpin RNA knockdown of SCN8A. These results demonstrate that a variant of NaV1.6 participates in the control of podosome and invadopodia formation and suggest that intracellular sodium release mediated by NaV1.6 may regulate cellular invasion of macrophages and melanoma cells. |
doi_str_mv | 10.1074/jbc.M801892200 |
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They also are expressed in non-excitable cells such as macrophages and neoplastic cells. Previously, in macrophages, we demonstrated expression of SCN8A, the gene that encodes the channel NaV1.6, and intracellular localization of NaV1.6 to regions near F-actin bundles, particularly at areas of cell attachment. Here we show that a splice variant of NaV1.6 regulates cellular invasion through its effects on podosome and invadopodia formation in macrophages and melanoma cells. cDNA sequence analysis of SCN8A from THP-1 cells, a human monocyte-macrophage cell line, confirmed the expression of a full-length splice variant that lacks exon 18. Immunoelectron microscopy demonstrated NaV1.6-positive staining within the electron dense podosome rosette structure. Pharmacologic antagonism with tetrodotoxin (TTX) in differentiated THP-1 cells or absence of functional NaV1.6 through a naturally occurring mutation (med) in mouse peritoneal macrophages inhibited podosome formation. Agonist-mediated activation of the channel with veratridine caused release of sodium from cationic vesicular compartments, uptake by mitochondria, and mitochondrial calcium release through the Na/Ca exchanger. Invasion by differentiated THP-1 and HTB-66 cells, an invasive melanoma cell line, through extracellular matrix was inhibited by TTX. THP-1 invasion also was inhibited by small hairpin RNA knockdown of SCN8A. These results demonstrate that a variant of NaV1.6 participates in the control of podosome and invadopodia formation and suggest that intracellular sodium release mediated by NaV1.6 may regulate cellular invasion of macrophages and melanoma cells.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M801892200</identifier><identifier>PMID: 19136557</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Alternative Splicing ; Animals ; Calcium - metabolism ; Cell Differentiation - drug effects ; Cell Line, Tumor ; Cell Membrane Structures - metabolism ; Exons ; Humans ; Macrophages, Peritoneal - metabolism ; Macrophages, Peritoneal - pathology ; Melanoma - metabolism ; Melanoma - pathology ; Mice ; Mice, Mutant Strains ; Mitochondria - metabolism ; Mutation ; NAV1.6 Voltage-Gated Sodium Channel ; Neoplasm Invasiveness ; Nerve Tissue Proteins - metabolism ; Sodium Channel Blockers - pharmacology ; Sodium Channels - metabolism ; Tetrodotoxin - pharmacology ; Veratridine - pharmacology</subject><ispartof>The Journal of biological chemistry, 2009-03, Vol.284 (12), p.8114-8126</ispartof><rights>2009 © 2009 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c465t-6d123c726c7febd3a42cf3b03e350594f93ba7b2ffe3351d6cbe68c8a213d0ad3</citedby><cites>FETCH-LOGICAL-c465t-6d123c726c7febd3a42cf3b03e350594f93ba7b2ffe3351d6cbe68c8a213d0ad3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19136557$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Carrithers, Michael D.</creatorcontrib><creatorcontrib>Chatterjee, Gouri</creatorcontrib><creatorcontrib>Carrithers, Lisette M.</creatorcontrib><creatorcontrib>Offoha, Roosevelt</creatorcontrib><creatorcontrib>Iheagwara, Uzoma</creatorcontrib><creatorcontrib>Rahner, Christoph</creatorcontrib><creatorcontrib>Graham, Morven</creatorcontrib><creatorcontrib>Waxman, Stephen G.</creatorcontrib><title>Regulation of Podosome Formation in Macrophages by a Splice Variant of the Sodium Channel SCN8A</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Voltage-gated sodium channels initiate electrical signaling in excitable cells such as muscle and neurons. They also are expressed in non-excitable cells such as macrophages and neoplastic cells. Previously, in macrophages, we demonstrated expression of SCN8A, the gene that encodes the channel NaV1.6, and intracellular localization of NaV1.6 to regions near F-actin bundles, particularly at areas of cell attachment. Here we show that a splice variant of NaV1.6 regulates cellular invasion through its effects on podosome and invadopodia formation in macrophages and melanoma cells. cDNA sequence analysis of SCN8A from THP-1 cells, a human monocyte-macrophage cell line, confirmed the expression of a full-length splice variant that lacks exon 18. Immunoelectron microscopy demonstrated NaV1.6-positive staining within the electron dense podosome rosette structure. Pharmacologic antagonism with tetrodotoxin (TTX) in differentiated THP-1 cells or absence of functional NaV1.6 through a naturally occurring mutation (med) in mouse peritoneal macrophages inhibited podosome formation. Agonist-mediated activation of the channel with veratridine caused release of sodium from cationic vesicular compartments, uptake by mitochondria, and mitochondrial calcium release through the Na/Ca exchanger. Invasion by differentiated THP-1 and HTB-66 cells, an invasive melanoma cell line, through extracellular matrix was inhibited by TTX. THP-1 invasion also was inhibited by small hairpin RNA knockdown of SCN8A. These results demonstrate that a variant of NaV1.6 participates in the control of podosome and invadopodia formation and suggest that intracellular sodium release mediated by NaV1.6 may regulate cellular invasion of macrophages and melanoma cells.</description><subject>Alternative Splicing</subject><subject>Animals</subject><subject>Calcium - metabolism</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Line, Tumor</subject><subject>Cell Membrane Structures - metabolism</subject><subject>Exons</subject><subject>Humans</subject><subject>Macrophages, Peritoneal - metabolism</subject><subject>Macrophages, Peritoneal - pathology</subject><subject>Melanoma - metabolism</subject><subject>Melanoma - pathology</subject><subject>Mice</subject><subject>Mice, Mutant Strains</subject><subject>Mitochondria - metabolism</subject><subject>Mutation</subject><subject>NAV1.6 Voltage-Gated Sodium Channel</subject><subject>Neoplasm Invasiveness</subject><subject>Nerve Tissue Proteins - metabolism</subject><subject>Sodium Channel Blockers - pharmacology</subject><subject>Sodium Channels - metabolism</subject><subject>Tetrodotoxin - pharmacology</subject><subject>Veratridine - pharmacology</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1v1DAQhi0EokvhyhEsIXHL4o84cY7VigJSC4iliJvlj_HGVRJv7QTUf4-rrNQTcxlp9MyrmQeh15RsKWnrD7fGbq8lobJjjJAnaEOJ5BUX9PdTtCGE0apjQp6hFznfklJ1R5-jM9pR3gjRbpD6AYdl0HOIE44ef48u5jgCvoxpXKdhwtfapnjs9QEyNvdY4_1xCBbwL52CnuaHxbkHvI8uLCPe9XqaYMD73Vd58RI983rI8OrUz9HN5cefu8_V1bdPX3YXV5WtGzFXjaOM25Y1tvVgHNc1s54bwoELIrrad9zo1jDvgZfnXGMNNNJKzSh3RDt-jt6vuccU7xbIsxpDtjAMeoK4ZMWKFyY6WcDtCpaXck7g1TGFUad7RYl6UKqKUvWotCy8OSUvZgT3iJ8cFuDdCvTh0P8NCZQJ0fYwKiZrRZmSlNaFertSXkelDylkdbNnhHJCGyKamhVCrgQUT38CJJVtgMmCK5l2Vi6G_934Dw8LmR4</recordid><startdate>20090320</startdate><enddate>20090320</enddate><creator>Carrithers, Michael D.</creator><creator>Chatterjee, Gouri</creator><creator>Carrithers, Lisette M.</creator><creator>Offoha, Roosevelt</creator><creator>Iheagwara, Uzoma</creator><creator>Rahner, Christoph</creator><creator>Graham, Morven</creator><creator>Waxman, Stephen G.</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T5</scope><scope>H94</scope></search><sort><creationdate>20090320</creationdate><title>Regulation of Podosome Formation in Macrophages by a Splice Variant of the Sodium Channel SCN8A</title><author>Carrithers, Michael D. ; Chatterjee, Gouri ; Carrithers, Lisette M. ; Offoha, Roosevelt ; Iheagwara, Uzoma ; Rahner, Christoph ; Graham, Morven ; Waxman, Stephen G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c465t-6d123c726c7febd3a42cf3b03e350594f93ba7b2ffe3351d6cbe68c8a213d0ad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Alternative Splicing</topic><topic>Animals</topic><topic>Calcium - metabolism</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Line, Tumor</topic><topic>Cell Membrane Structures - metabolism</topic><topic>Exons</topic><topic>Humans</topic><topic>Macrophages, Peritoneal - metabolism</topic><topic>Macrophages, Peritoneal - pathology</topic><topic>Melanoma - metabolism</topic><topic>Melanoma - pathology</topic><topic>Mice</topic><topic>Mice, Mutant Strains</topic><topic>Mitochondria - metabolism</topic><topic>Mutation</topic><topic>NAV1.6 Voltage-Gated Sodium Channel</topic><topic>Neoplasm Invasiveness</topic><topic>Nerve Tissue Proteins - metabolism</topic><topic>Sodium Channel Blockers - pharmacology</topic><topic>Sodium Channels - metabolism</topic><topic>Tetrodotoxin - pharmacology</topic><topic>Veratridine - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Carrithers, Michael D.</creatorcontrib><creatorcontrib>Chatterjee, Gouri</creatorcontrib><creatorcontrib>Carrithers, Lisette M.</creatorcontrib><creatorcontrib>Offoha, Roosevelt</creatorcontrib><creatorcontrib>Iheagwara, Uzoma</creatorcontrib><creatorcontrib>Rahner, Christoph</creatorcontrib><creatorcontrib>Graham, Morven</creatorcontrib><creatorcontrib>Waxman, Stephen G.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Carrithers, Michael D.</au><au>Chatterjee, Gouri</au><au>Carrithers, Lisette M.</au><au>Offoha, Roosevelt</au><au>Iheagwara, Uzoma</au><au>Rahner, Christoph</au><au>Graham, Morven</au><au>Waxman, Stephen G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Regulation of Podosome Formation in Macrophages by a Splice Variant of the Sodium Channel SCN8A</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2009-03-20</date><risdate>2009</risdate><volume>284</volume><issue>12</issue><spage>8114</spage><epage>8126</epage><pages>8114-8126</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Voltage-gated sodium channels initiate electrical signaling in excitable cells such as muscle and neurons. They also are expressed in non-excitable cells such as macrophages and neoplastic cells. Previously, in macrophages, we demonstrated expression of SCN8A, the gene that encodes the channel NaV1.6, and intracellular localization of NaV1.6 to regions near F-actin bundles, particularly at areas of cell attachment. Here we show that a splice variant of NaV1.6 regulates cellular invasion through its effects on podosome and invadopodia formation in macrophages and melanoma cells. cDNA sequence analysis of SCN8A from THP-1 cells, a human monocyte-macrophage cell line, confirmed the expression of a full-length splice variant that lacks exon 18. Immunoelectron microscopy demonstrated NaV1.6-positive staining within the electron dense podosome rosette structure. Pharmacologic antagonism with tetrodotoxin (TTX) in differentiated THP-1 cells or absence of functional NaV1.6 through a naturally occurring mutation (med) in mouse peritoneal macrophages inhibited podosome formation. Agonist-mediated activation of the channel with veratridine caused release of sodium from cationic vesicular compartments, uptake by mitochondria, and mitochondrial calcium release through the Na/Ca exchanger. Invasion by differentiated THP-1 and HTB-66 cells, an invasive melanoma cell line, through extracellular matrix was inhibited by TTX. THP-1 invasion also was inhibited by small hairpin RNA knockdown of SCN8A. These results demonstrate that a variant of NaV1.6 participates in the control of podosome and invadopodia formation and suggest that intracellular sodium release mediated by NaV1.6 may regulate cellular invasion of macrophages and melanoma cells.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>19136557</pmid><doi>10.1074/jbc.M801892200</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Alternative Splicing Animals Calcium - metabolism Cell Differentiation - drug effects Cell Line, Tumor Cell Membrane Structures - metabolism Exons Humans Macrophages, Peritoneal - metabolism Macrophages, Peritoneal - pathology Melanoma - metabolism Melanoma - pathology Mice Mice, Mutant Strains Mitochondria - metabolism Mutation NAV1.6 Voltage-Gated Sodium Channel Neoplasm Invasiveness Nerve Tissue Proteins - metabolism Sodium Channel Blockers - pharmacology Sodium Channels - metabolism Tetrodotoxin - pharmacology Veratridine - pharmacology |
title | Regulation of Podosome Formation in Macrophages by a Splice Variant of the Sodium Channel SCN8A |
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