Morphological Differentiation of N1E-115 Neuroblastoma Cells by Dimethyl Sulfoxide Activation of Lipid Second Messengers
Quantitative changes in the lipid second messenger diacylglycerol (DAG) were studied in the rat neuroblastoma N1E-115 following exposure to the differentiating agent dimethylsulfoxide (DMSO). Relatively high basal levels of DAG are present in these cells, and addition of 2% DMSO elicited a biphasic...
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Veröffentlicht in: | Experimental cell research 1996-04, Vol.224 (1), p.16-27 |
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description | Quantitative changes in the lipid second messenger diacylglycerol (DAG) were studied in the rat neuroblastoma N1E-115 following exposure to the differentiating agent dimethylsulfoxide (DMSO). Relatively high basal levels of DAG are present in these cells, and addition of 2% DMSO elicited a biphasic increase in DAG levels, dependent on the presence of extracellular Ca2+. Exposure to DMSO also elicited a rapid increase in inositol phosphate and a slight increase in phosphatidic acid (PA), trailing that of DAG. The molecular species (MS) of DAG were analyzed. Within 60 s of DMSO application there were transient increases of DAG representative of phosphatidylinositol (PI) hydrolysis. At longer intervals, more DAG originated from phosphatidylcholine. The MS composition of newly formed PA resembled that of PI and native DAG. Inhibition studies indicated that DAG is formed in the DMSO-treated cells by phospholipases C and that PA formed later is a result of DAG phosphorylation and not activity of phospholipase D (PLD). Undifferentiated cells exhibited an active PLD pathway. In contrast, PLD in DMSO-differentiated cells was not active. In examining the involvement of the sphingomyelin pathway, DMSO exposure was found to increase ceramide levels with a concomitant decrease in sphingomyelin. Addition of the exogenous, soluble analog C6-ceramide to undifferentiated cells resulted in dramatic reductions in DAG and PA levels and PLD activity. These results indicate that DMSO treatment inactivates PLD while activating phospholipases C and the sphingomyelin pathway, suggesting a “switch” between signal transduction pathways in the undifferentiated and differentiated states of N1E-115. |
doi_str_mv | 10.1006/excr.1996.0107 |
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Relatively high basal levels of DAG are present in these cells, and addition of 2% DMSO elicited a biphasic increase in DAG levels, dependent on the presence of extracellular Ca2+. Exposure to DMSO also elicited a rapid increase in inositol phosphate and a slight increase in phosphatidic acid (PA), trailing that of DAG. The molecular species (MS) of DAG were analyzed. Within 60 s of DMSO application there were transient increases of DAG representative of phosphatidylinositol (PI) hydrolysis. At longer intervals, more DAG originated from phosphatidylcholine. The MS composition of newly formed PA resembled that of PI and native DAG. Inhibition studies indicated that DAG is formed in the DMSO-treated cells by phospholipases C and that PA formed later is a result of DAG phosphorylation and not activity of phospholipase D (PLD). Undifferentiated cells exhibited an active PLD pathway. In contrast, PLD in DMSO-differentiated cells was not active. In examining the involvement of the sphingomyelin pathway, DMSO exposure was found to increase ceramide levels with a concomitant decrease in sphingomyelin. Addition of the exogenous, soluble analog C6-ceramide to undifferentiated cells resulted in dramatic reductions in DAG and PA levels and PLD activity. These results indicate that DMSO treatment inactivates PLD while activating phospholipases C and the sphingomyelin pathway, suggesting a “switch” between signal transduction pathways in the undifferentiated and differentiated states of N1E-115.</description><identifier>ISSN: 0014-4827</identifier><identifier>EISSN: 1090-2422</identifier><identifier>DOI: 10.1006/excr.1996.0107</identifier><identifier>PMID: 8612681</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Calcium - metabolism ; Cell Differentiation ; Ceramides - metabolism ; Diacylglycerol Kinase ; Diglycerides - metabolism ; Dimethyl Sulfoxide - pharmacology ; Enzyme Inhibitors - pharmacology ; Fatty Acids - analysis ; Inositol 1,4,5-Trisphosphate - metabolism ; Neuroblastoma ; Neurons - cytology ; Neurons - physiology ; Phosphatidate Phosphatase - antagonists & inhibitors ; Phosphatidic Acids - metabolism ; Phospholipase D - analysis ; Phosphotransferases (Alcohol Group Acceptor) - antagonists & inhibitors ; Second Messenger Systems ; Sphingomyelins - metabolism ; Tumor Cells, Cultured</subject><ispartof>Experimental cell research, 1996-04, Vol.224 (1), p.16-27</ispartof><rights>1996 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c339t-7e0cfcb35515bc9e0879c6e5d51c8481ba6d8de0d75a07889764e39927784cf53</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S001448279690107X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8612681$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Clejan, Sanda</creatorcontrib><creatorcontrib>Dotson, Robert S.</creatorcontrib><creatorcontrib>Wolf, Erich W.</creatorcontrib><creatorcontrib>Corb, Michael P.</creatorcontrib><creatorcontrib>Ide, Charles F.</creatorcontrib><title>Morphological Differentiation of N1E-115 Neuroblastoma Cells by Dimethyl Sulfoxide Activation of Lipid Second Messengers</title><title>Experimental cell research</title><addtitle>Exp Cell Res</addtitle><description>Quantitative changes in the lipid second messenger diacylglycerol (DAG) were studied in the rat neuroblastoma N1E-115 following exposure to the differentiating agent dimethylsulfoxide (DMSO). Relatively high basal levels of DAG are present in these cells, and addition of 2% DMSO elicited a biphasic increase in DAG levels, dependent on the presence of extracellular Ca2+. Exposure to DMSO also elicited a rapid increase in inositol phosphate and a slight increase in phosphatidic acid (PA), trailing that of DAG. The molecular species (MS) of DAG were analyzed. Within 60 s of DMSO application there were transient increases of DAG representative of phosphatidylinositol (PI) hydrolysis. At longer intervals, more DAG originated from phosphatidylcholine. The MS composition of newly formed PA resembled that of PI and native DAG. Inhibition studies indicated that DAG is formed in the DMSO-treated cells by phospholipases C and that PA formed later is a result of DAG phosphorylation and not activity of phospholipase D (PLD). Undifferentiated cells exhibited an active PLD pathway. In contrast, PLD in DMSO-differentiated cells was not active. In examining the involvement of the sphingomyelin pathway, DMSO exposure was found to increase ceramide levels with a concomitant decrease in sphingomyelin. Addition of the exogenous, soluble analog C6-ceramide to undifferentiated cells resulted in dramatic reductions in DAG and PA levels and PLD activity. These results indicate that DMSO treatment inactivates PLD while activating phospholipases C and the sphingomyelin pathway, suggesting a “switch” between signal transduction pathways in the undifferentiated and differentiated states of N1E-115.</description><subject>Calcium - metabolism</subject><subject>Cell Differentiation</subject><subject>Ceramides - metabolism</subject><subject>Diacylglycerol Kinase</subject><subject>Diglycerides - metabolism</subject><subject>Dimethyl Sulfoxide - pharmacology</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Fatty Acids - analysis</subject><subject>Inositol 1,4,5-Trisphosphate - metabolism</subject><subject>Neuroblastoma</subject><subject>Neurons - cytology</subject><subject>Neurons - physiology</subject><subject>Phosphatidate Phosphatase - antagonists & inhibitors</subject><subject>Phosphatidic Acids - metabolism</subject><subject>Phospholipase D - analysis</subject><subject>Phosphotransferases (Alcohol Group Acceptor) - antagonists & inhibitors</subject><subject>Second Messenger Systems</subject><subject>Sphingomyelins - metabolism</subject><subject>Tumor Cells, Cultured</subject><issn>0014-4827</issn><issn>1090-2422</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kDtv2zAUhYmgQeqkWbsV4NRNzqUefIyB67QBnGRIOhMUeRWzkESXlAz730eGDW-Z7nAeOPcj5DuDOQPgd7izcc6U4nNgIC7IjIGCLC_z_AuZAbAyK2UuvpLrlP4BgJSMX5EryVnOJZuR3VOIm3Vow7u3pqW_fNNgxH7wZvChp6Ghz2yZMVbRZxxjqFuThtAZusC2TbTeT4kOh_W-pa9j24Sdd0jv7eC35_zKb7yjr2hD7-gTpoT9O8b0jVw2pk14e7o35O_D8m3xJ1u9_H5c3K8yWxRqyASCbWxdVBWraqsQpFCWY-UqZmUpWW24kw7BicqAkFIJXmKhVC6ELG1TFTfk57F3E8P_EdOgO5_stN70GMakhVBScgGTcX402hhSitjoTfSdiXvNQB9Q6wNqfUCtD6inwI9T81h36M72E9tJl0cdp_e2HqNO1mNv0fmIdtAu-M-qPwBbKY5B</recordid><startdate>19960410</startdate><enddate>19960410</enddate><creator>Clejan, Sanda</creator><creator>Dotson, Robert S.</creator><creator>Wolf, Erich W.</creator><creator>Corb, Michael P.</creator><creator>Ide, Charles F.</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>19960410</creationdate><title>Morphological Differentiation of N1E-115 Neuroblastoma Cells by Dimethyl Sulfoxide Activation of Lipid Second Messengers</title><author>Clejan, Sanda ; Dotson, Robert S. ; Wolf, Erich W. ; Corb, Michael P. ; Ide, Charles F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c339t-7e0cfcb35515bc9e0879c6e5d51c8481ba6d8de0d75a07889764e39927784cf53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Calcium - metabolism</topic><topic>Cell Differentiation</topic><topic>Ceramides - metabolism</topic><topic>Diacylglycerol Kinase</topic><topic>Diglycerides - metabolism</topic><topic>Dimethyl Sulfoxide - pharmacology</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Fatty Acids - analysis</topic><topic>Inositol 1,4,5-Trisphosphate - metabolism</topic><topic>Neuroblastoma</topic><topic>Neurons - cytology</topic><topic>Neurons - physiology</topic><topic>Phosphatidate Phosphatase - antagonists & inhibitors</topic><topic>Phosphatidic Acids - metabolism</topic><topic>Phospholipase D - analysis</topic><topic>Phosphotransferases (Alcohol Group Acceptor) - antagonists & inhibitors</topic><topic>Second Messenger Systems</topic><topic>Sphingomyelins - metabolism</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Clejan, Sanda</creatorcontrib><creatorcontrib>Dotson, Robert S.</creatorcontrib><creatorcontrib>Wolf, Erich W.</creatorcontrib><creatorcontrib>Corb, Michael P.</creatorcontrib><creatorcontrib>Ide, Charles F.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental cell research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Clejan, Sanda</au><au>Dotson, Robert S.</au><au>Wolf, Erich W.</au><au>Corb, Michael P.</au><au>Ide, Charles F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Morphological Differentiation of N1E-115 Neuroblastoma Cells by Dimethyl Sulfoxide Activation of Lipid Second Messengers</atitle><jtitle>Experimental cell research</jtitle><addtitle>Exp Cell Res</addtitle><date>1996-04-10</date><risdate>1996</risdate><volume>224</volume><issue>1</issue><spage>16</spage><epage>27</epage><pages>16-27</pages><issn>0014-4827</issn><eissn>1090-2422</eissn><abstract>Quantitative changes in the lipid second messenger diacylglycerol (DAG) were studied in the rat neuroblastoma N1E-115 following exposure to the differentiating agent dimethylsulfoxide (DMSO). Relatively high basal levels of DAG are present in these cells, and addition of 2% DMSO elicited a biphasic increase in DAG levels, dependent on the presence of extracellular Ca2+. Exposure to DMSO also elicited a rapid increase in inositol phosphate and a slight increase in phosphatidic acid (PA), trailing that of DAG. The molecular species (MS) of DAG were analyzed. Within 60 s of DMSO application there were transient increases of DAG representative of phosphatidylinositol (PI) hydrolysis. At longer intervals, more DAG originated from phosphatidylcholine. The MS composition of newly formed PA resembled that of PI and native DAG. Inhibition studies indicated that DAG is formed in the DMSO-treated cells by phospholipases C and that PA formed later is a result of DAG phosphorylation and not activity of phospholipase D (PLD). Undifferentiated cells exhibited an active PLD pathway. In contrast, PLD in DMSO-differentiated cells was not active. In examining the involvement of the sphingomyelin pathway, DMSO exposure was found to increase ceramide levels with a concomitant decrease in sphingomyelin. Addition of the exogenous, soluble analog C6-ceramide to undifferentiated cells resulted in dramatic reductions in DAG and PA levels and PLD activity. These results indicate that DMSO treatment inactivates PLD while activating phospholipases C and the sphingomyelin pathway, suggesting a “switch” between signal transduction pathways in the undifferentiated and differentiated states of N1E-115.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>8612681</pmid><doi>10.1006/excr.1996.0107</doi><tpages>12</tpages></addata></record> |
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subjects | Calcium - metabolism Cell Differentiation Ceramides - metabolism Diacylglycerol Kinase Diglycerides - metabolism Dimethyl Sulfoxide - pharmacology Enzyme Inhibitors - pharmacology Fatty Acids - analysis Inositol 1,4,5-Trisphosphate - metabolism Neuroblastoma Neurons - cytology Neurons - physiology Phosphatidate Phosphatase - antagonists & inhibitors Phosphatidic Acids - metabolism Phospholipase D - analysis Phosphotransferases (Alcohol Group Acceptor) - antagonists & inhibitors Second Messenger Systems Sphingomyelins - metabolism Tumor Cells, Cultured |
title | Morphological Differentiation of N1E-115 Neuroblastoma Cells by Dimethyl Sulfoxide Activation of Lipid Second Messengers |
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