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Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices
The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extrac...
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Published in: | Proceedings of the National Academy of Sciences - PNAS 1978-10, Vol.75 (10), p.5195-5199 |
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description | The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extraction. When the slices were incubated in a standard Krebs-Ringer solution, proteins Ia and Ib were present almost entirely in the dephosphorylated form. Incubation with cyclic AMP, 8-bromo cyclic AMP, N6-monobutyryl cyclic AMP, or with a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the phosphorylation of proteins Ia and Ib in the slices. Depolarization of neuronal membranes by high K+ or by veratridine was also associated with an increased phosphorylation of proteins Ia and Ib. The effect of depolarizing agents, but not that of cyclic nucleotides or 3-isobutyl-1-methylxanthine, required the presence of external Ca2+ in the incubation medium. Tetrodotoxin blocked the stimulation of the phosphorylation of proteins Ia and Ib induced by veratridine but not that induced by the other agents tested. Incubation of the brain slices with 8-bromo cyclic AMP, 3-isobutyl-1-methylxanthine, high K+, or veratridine also increased the state of phosphorylation of two other neuronal proteins found in extracts of the slices. |
doi_str_mv | 10.1073/pnas.75.10.5195 |
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For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extraction. When the slices were incubated in a standard Krebs-Ringer solution, proteins Ia and Ib were present almost entirely in the dephosphorylated form. Incubation with cyclic AMP, 8-bromo cyclic AMP, N6-monobutyryl cyclic AMP, or with a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the phosphorylation of proteins Ia and Ib in the slices. Depolarization of neuronal membranes by high K+ or by veratridine was also associated with an increased phosphorylation of proteins Ia and Ib. The effect of depolarizing agents, but not that of cyclic nucleotides or 3-isobutyl-1-methylxanthine, required the presence of external Ca2+ in the incubation medium. Tetrodotoxin blocked the stimulation of the phosphorylation of proteins Ia and Ib induced by veratridine but not that induced by the other agents tested. Incubation of the brain slices with 8-bromo cyclic AMP, 3-isobutyl-1-methylxanthine, high K+, or veratridine also increased the state of phosphorylation of two other neuronal proteins found in extracts of the slices.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.75.10.5195</identifier><identifier>PMID: 84386</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>1-Methyl-3-isobutylxanthine - pharmacology ; Animals ; Brain ; Calcium ; Calcium - metabolism ; Cerebral cortex ; Cerebral Cortex - metabolism ; Cyclic AMP - physiology ; Cyclic nucleotides ; Gels ; Homogenization ; In Vitro Techniques ; Nerve Tissue Proteins - metabolism ; Neurons ; Phosphatases ; Phosphorylation ; Potassium - pharmacology ; Rats ; Tetrodotoxin ; Tetrodotoxin - pharmacology ; Veratridine - pharmacology</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1978-10, Vol.75 (10), p.5195-5199</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c459t-414c0588b5519a5786d2eccde540aa43ad3e9875852ead917019cd5358c1606a3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/75/10.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/69088$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/69088$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793,58238,58471</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/84386$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Forn, Javier</creatorcontrib><creatorcontrib>Greengard, Paul</creatorcontrib><title>Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extraction. When the slices were incubated in a standard Krebs-Ringer solution, proteins Ia and Ib were present almost entirely in the dephosphorylated form. Incubation with cyclic AMP, 8-bromo cyclic AMP, N6-monobutyryl cyclic AMP, or with a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the phosphorylation of proteins Ia and Ib in the slices. Depolarization of neuronal membranes by high K+ or by veratridine was also associated with an increased phosphorylation of proteins Ia and Ib. The effect of depolarizing agents, but not that of cyclic nucleotides or 3-isobutyl-1-methylxanthine, required the presence of external Ca2+ in the incubation medium. Tetrodotoxin blocked the stimulation of the phosphorylation of proteins Ia and Ib induced by veratridine but not that induced by the other agents tested. Incubation of the brain slices with 8-bromo cyclic AMP, 3-isobutyl-1-methylxanthine, high K+, or veratridine also increased the state of phosphorylation of two other neuronal proteins found in extracts of the slices.</description><subject>1-Methyl-3-isobutylxanthine - pharmacology</subject><subject>Animals</subject><subject>Brain</subject><subject>Calcium</subject><subject>Calcium - metabolism</subject><subject>Cerebral cortex</subject><subject>Cerebral Cortex - metabolism</subject><subject>Cyclic AMP - physiology</subject><subject>Cyclic nucleotides</subject><subject>Gels</subject><subject>Homogenization</subject><subject>In Vitro Techniques</subject><subject>Nerve Tissue Proteins - metabolism</subject><subject>Neurons</subject><subject>Phosphatases</subject><subject>Phosphorylation</subject><subject>Potassium - pharmacology</subject><subject>Rats</subject><subject>Tetrodotoxin</subject><subject>Tetrodotoxin - pharmacology</subject><subject>Veratridine - pharmacology</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1978</creationdate><recordtype>article</recordtype><recordid>eNp9kc9vFCEUx4mx0bV6NtHEcPM0WxhgBg4emvFn0mjT6pmwzJtdmilMgDGuV_9xGbdu7MUTeXy_Hx7vfRF6TsmakpadTd6kdStKsRZUiQdoRYmiVcMVeYhWhNRtJXnNH6MnKd0QQpSQ5BE6kZzJZoV-vYUpjCa6n85v8fkWfE7Y-B53ezs6iz_PdoSQXQ8JX8F2Hk0GnHeAL3chTbsQ9-XGBY_DgK8nsG5YIJhj8GbElzFkcD5h5_GVybiDCJtYhC7EDD_wdWkB6Sk6GcyY4NndeYq-vX_3tftYXXz58Kk7v6gsFypXnHJLhJQbUeY0opVNX4O1PQhOjOHM9AyUbIUUNZhe0ZZQZXvBhLS0IY1hp-jN4d1p3txCb8us5S96iu7WxL0Oxun7inc7vQ3fNWNNrerCnx14G0NKEYYjSolestBLFroVS71kUYiX_3Y8-v8sv6iv7tQF-6vdw1__16CHeRzLHnNxvjg4b1IO8WhtFJGS_Qb3uamj</recordid><startdate>19781001</startdate><enddate>19781001</enddate><creator>Forn, Javier</creator><creator>Greengard, Paul</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><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>5PM</scope></search><sort><creationdate>19781001</creationdate><title>Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices</title><author>Forn, Javier ; Greengard, Paul</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c459t-414c0588b5519a5786d2eccde540aa43ad3e9875852ead917019cd5358c1606a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1978</creationdate><topic>1-Methyl-3-isobutylxanthine - pharmacology</topic><topic>Animals</topic><topic>Brain</topic><topic>Calcium</topic><topic>Calcium - metabolism</topic><topic>Cerebral cortex</topic><topic>Cerebral Cortex - metabolism</topic><topic>Cyclic AMP - physiology</topic><topic>Cyclic nucleotides</topic><topic>Gels</topic><topic>Homogenization</topic><topic>In Vitro Techniques</topic><topic>Nerve Tissue Proteins - metabolism</topic><topic>Neurons</topic><topic>Phosphatases</topic><topic>Phosphorylation</topic><topic>Potassium - pharmacology</topic><topic>Rats</topic><topic>Tetrodotoxin</topic><topic>Tetrodotoxin - pharmacology</topic><topic>Veratridine - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Forn, Javier</creatorcontrib><creatorcontrib>Greengard, Paul</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Forn, Javier</au><au>Greengard, Paul</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1978-10-01</date><risdate>1978</risdate><volume>75</volume><issue>10</issue><spage>5195</spage><epage>5199</epage><pages>5195-5199</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>The regulation of the state of phosphorylation of two specific neuronal proteins, designated protein Ia and protein Ib, has been studied in slices of rat cerebral cortex incubated in vitro. For this purpose, a method was developed that prevents dephosphorylation of these proteins during their extraction. When the slices were incubated in a standard Krebs-Ringer solution, proteins Ia and Ib were present almost entirely in the dephosphorylated form. Incubation with cyclic AMP, 8-bromo cyclic AMP, N6-monobutyryl cyclic AMP, or with a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the phosphorylation of proteins Ia and Ib in the slices. Depolarization of neuronal membranes by high K+ or by veratridine was also associated with an increased phosphorylation of proteins Ia and Ib. The effect of depolarizing agents, but not that of cyclic nucleotides or 3-isobutyl-1-methylxanthine, required the presence of external Ca2+ in the incubation medium. Tetrodotoxin blocked the stimulation of the phosphorylation of proteins Ia and Ib induced by veratridine but not that induced by the other agents tested. Incubation of the brain slices with 8-bromo cyclic AMP, 3-isobutyl-1-methylxanthine, high K+, or veratridine also increased the state of phosphorylation of two other neuronal proteins found in extracts of the slices.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>84386</pmid><doi>10.1073/pnas.75.10.5195</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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subjects | 1-Methyl-3-isobutylxanthine - pharmacology Animals Brain Calcium Calcium - metabolism Cerebral cortex Cerebral Cortex - metabolism Cyclic AMP - physiology Cyclic nucleotides Gels Homogenization In Vitro Techniques Nerve Tissue Proteins - metabolism Neurons Phosphatases Phosphorylation Potassium - pharmacology Rats Tetrodotoxin Tetrodotoxin - pharmacology Veratridine - pharmacology |
title | Depolarizing Agents and Cyclic Nucleotides Regulate the Phosphorylation of Specific Neuronal Proteins in Rat Cerebral Cortex Slices |
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