Real-time measurement of free Ca2+ changes in CNS myelin by two-photon microscopy
Here we describe a technique for measuring changes in Ca 2+ in the cytosolic domain of mature compact myelin of live axons in the central nervous system (CNS). We label the myelin sheath of optic nerve and dorsal column axons by using the Ca 2+ indicator X-rhod-1 coupled with DiOC 6 (3) to produce b...
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| Veröffentlicht in: | Nature medicine 2007-07, Vol.13 (7), p.874-879 |
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| creator | Micu, Ileana Ridsdale, Andrew Zhang, Lingqing Woulfe, John McClintock, Jeff Brantner, Christine A Andrews, S Brian Stys, Peter K |
| description | Here we describe a technique for measuring changes in Ca
2+
in the cytosolic domain of mature compact myelin of live axons in the central nervous system (CNS). We label the myelin sheath of optic nerve and dorsal column axons by using the Ca
2+
indicator X-rhod-1 coupled with DiOC
6
(3) to produce bright myelin counterstaining, thereby providing unambiguous identification of the myelin sheath for analysis of two-photon excited fluorescence. We present evidence for localization of the Ca
2+
reporter to the cytosolic domain of myelin, obtained by using fluorescence lifetime, spectral measurements and Mn
2+
quenching. Chemical ischemia increased myelinic X-rhod-1 fluorescence (∼50% after 30 min) in a manner dependent on extracellular Ca
2+
. Inhibiting Na
+
-dependent glutamate transporters (with TBOA) or glycine transporters (with sarcosine and ALX-1393) reduced the ischemia-induced increase in Ca
2+
. We show that myelinic
N
-methyl-
D
-aspartate (NMDA) receptors are activated by the two conventional coagonists glutamate and glycine, which are released by specific transporters under conditions of cellular Na
+
loading and depolarization in injured white matter. This new technique facilitates detailed studies of living myelin, a vital component of the mammalian CNS. |
| doi_str_mv | 10.1038/nm1568 |
| format | Article |
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2+
in the cytosolic domain of mature compact myelin of live axons in the central nervous system (CNS). We label the myelin sheath of optic nerve and dorsal column axons by using the Ca
2+
indicator X-rhod-1 coupled with DiOC
6
(3) to produce bright myelin counterstaining, thereby providing unambiguous identification of the myelin sheath for analysis of two-photon excited fluorescence. We present evidence for localization of the Ca
2+
reporter to the cytosolic domain of myelin, obtained by using fluorescence lifetime, spectral measurements and Mn
2+
quenching. Chemical ischemia increased myelinic X-rhod-1 fluorescence (∼50% after 30 min) in a manner dependent on extracellular Ca
2+
. Inhibiting Na
+
-dependent glutamate transporters (with TBOA) or glycine transporters (with sarcosine and ALX-1393) reduced the ischemia-induced increase in Ca
2+
. We show that myelinic
N
-methyl-
D
-aspartate (NMDA) receptors are activated by the two conventional coagonists glutamate and glycine, which are released by specific transporters under conditions of cellular Na
+
loading and depolarization in injured white matter. This new technique facilitates detailed studies of living myelin, a vital component of the mammalian CNS.</description><identifier>ISSN: 1078-8956</identifier><identifier>EISSN: 1546-170X</identifier><identifier>DOI: 10.1038/nm1568</identifier><identifier>PMID: 17603496</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>Animals ; Biomedical and Life Sciences ; Biomedicine ; Calcium - metabolism ; Calcium Signaling ; Cancer Research ; Central nervous system ; Central Nervous System - cytology ; Central Nervous System - metabolism ; Fluorescence ; Fluorescent Dyes ; Infectious Diseases ; Mammals ; Metabolic Diseases ; Microscopy ; Molecular Medicine ; Myelin Sheath - metabolism ; Nervous system ; Neurons - cytology ; Neurosciences ; Rats ; Rats, Long-Evans ; technical-report ; Time Factors</subject><ispartof>Nature medicine, 2007-07, Vol.13 (7), p.874-879</ispartof><rights>Springer Nature America, Inc. 2007</rights><rights>COPYRIGHT 2007 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Jul 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c508t-7aed3a17afb49973584fabc2d929383dc97a5580b374d7e33bd3efc728792c7a3</citedby><cites>FETCH-LOGICAL-c508t-7aed3a17afb49973584fabc2d929383dc97a5580b374d7e33bd3efc728792c7a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nm1568$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nm1568$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17603496$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Micu, Ileana</creatorcontrib><creatorcontrib>Ridsdale, Andrew</creatorcontrib><creatorcontrib>Zhang, Lingqing</creatorcontrib><creatorcontrib>Woulfe, John</creatorcontrib><creatorcontrib>McClintock, Jeff</creatorcontrib><creatorcontrib>Brantner, Christine A</creatorcontrib><creatorcontrib>Andrews, S Brian</creatorcontrib><creatorcontrib>Stys, Peter K</creatorcontrib><title>Real-time measurement of free Ca2+ changes in CNS myelin by two-photon microscopy</title><title>Nature medicine</title><addtitle>Nat Med</addtitle><addtitle>Nat Med</addtitle><description>Here we describe a technique for measuring changes in Ca
2+
in the cytosolic domain of mature compact myelin of live axons in the central nervous system (CNS). We label the myelin sheath of optic nerve and dorsal column axons by using the Ca
2+
indicator X-rhod-1 coupled with DiOC
6
(3) to produce bright myelin counterstaining, thereby providing unambiguous identification of the myelin sheath for analysis of two-photon excited fluorescence. We present evidence for localization of the Ca
2+
reporter to the cytosolic domain of myelin, obtained by using fluorescence lifetime, spectral measurements and Mn
2+
quenching. Chemical ischemia increased myelinic X-rhod-1 fluorescence (∼50% after 30 min) in a manner dependent on extracellular Ca
2+
. Inhibiting Na
+
-dependent glutamate transporters (with TBOA) or glycine transporters (with sarcosine and ALX-1393) reduced the ischemia-induced increase in Ca
2+
. We show that myelinic
N
-methyl-
D
-aspartate (NMDA) receptors are activated by the two conventional coagonists glutamate and glycine, which are released by specific transporters under conditions of cellular Na
+
loading and depolarization in injured white matter. This new technique facilitates detailed studies of living myelin, a vital component of the mammalian CNS.</description><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Calcium - metabolism</subject><subject>Calcium Signaling</subject><subject>Cancer Research</subject><subject>Central nervous system</subject><subject>Central Nervous System - cytology</subject><subject>Central Nervous System - metabolism</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes</subject><subject>Infectious Diseases</subject><subject>Mammals</subject><subject>Metabolic Diseases</subject><subject>Microscopy</subject><subject>Molecular Medicine</subject><subject>Myelin Sheath - metabolism</subject><subject>Nervous system</subject><subject>Neurons - cytology</subject><subject>Neurosciences</subject><subject>Rats</subject><subject>Rats, Long-Evans</subject><subject>technical-report</subject><subject>Time Factors</subject><issn>1078-8956</issn><issn>1546-170X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>BENPR</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqN0l9rFDEQAPAgiq1VP4IEHyoiW5PNZrN5LId_CsViq-JbyGYnd1s2yZlk0fv25riD2nIPkoeE5DdDmBmEXlJyRgnr3ntHeds9QseUN21FBfn5uJyJ6KpO8vYIPUvplhDCCJdP0REVLWGNbI_R12vQU5VHB9iBTnMEBz7jYLGNAHih63fYrLRfQsKjx4svN9htYCrHfoPz71CtVyEHj91oYkgmrDfP0ROrpwQv9vsJ-v7xw7fF5-ry6tPF4vyyMpx0uRIaBqap0LZvpBSMd43VvakHWUvWscFIoTnvSM9EMwhgrB8YWCPqTsjaCM1O0Oku7zqGXzOkrNyYDEyT9hDmpARpSyLKC3z9AN6GOfryN1XXjNK2bbao2qGlnkCN3oYctVmCh6in4MGO5fqcyrppBKOy-LMDvqwBSikOBry9F1BMhj95qeeU1MXN9f_bqx_37ek_dlW6mVcpTHMeg08H4bZRKYJV6zg6HTeKErWdIbWboQJf7es19w6GO7YfmgLe7EAqT2Uy4l1BH6T6C51kyV4</recordid><startdate>20070701</startdate><enddate>20070701</enddate><creator>Micu, Ileana</creator><creator>Ridsdale, Andrew</creator><creator>Zhang, Lingqing</creator><creator>Woulfe, John</creator><creator>McClintock, Jeff</creator><creator>Brantner, Christine A</creator><creator>Andrews, S Brian</creator><creator>Stys, Peter K</creator><general>Nature Publishing Group US</general><general>Nature Publishing Group</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>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20070701</creationdate><title>Real-time measurement of free Ca2+ changes in CNS myelin by two-photon microscopy</title><author>Micu, Ileana ; Ridsdale, Andrew ; Zhang, Lingqing ; Woulfe, John ; McClintock, Jeff ; Brantner, Christine A ; Andrews, S Brian ; Stys, Peter K</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c508t-7aed3a17afb49973584fabc2d929383dc97a5580b374d7e33bd3efc728792c7a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Animals</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Calcium - 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Academic</collection><jtitle>Nature medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Micu, Ileana</au><au>Ridsdale, Andrew</au><au>Zhang, Lingqing</au><au>Woulfe, John</au><au>McClintock, Jeff</au><au>Brantner, Christine A</au><au>Andrews, S Brian</au><au>Stys, Peter K</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Real-time measurement of free Ca2+ changes in CNS myelin by two-photon microscopy</atitle><jtitle>Nature medicine</jtitle><stitle>Nat Med</stitle><addtitle>Nat Med</addtitle><date>2007-07-01</date><risdate>2007</risdate><volume>13</volume><issue>7</issue><spage>874</spage><epage>879</epage><pages>874-879</pages><issn>1078-8956</issn><eissn>1546-170X</eissn><abstract>Here we describe a technique for measuring changes in Ca
2+
in the cytosolic domain of mature compact myelin of live axons in the central nervous system (CNS). We label the myelin sheath of optic nerve and dorsal column axons by using the Ca
2+
indicator X-rhod-1 coupled with DiOC
6
(3) to produce bright myelin counterstaining, thereby providing unambiguous identification of the myelin sheath for analysis of two-photon excited fluorescence. We present evidence for localization of the Ca
2+
reporter to the cytosolic domain of myelin, obtained by using fluorescence lifetime, spectral measurements and Mn
2+
quenching. Chemical ischemia increased myelinic X-rhod-1 fluorescence (∼50% after 30 min) in a manner dependent on extracellular Ca
2+
. Inhibiting Na
+
-dependent glutamate transporters (with TBOA) or glycine transporters (with sarcosine and ALX-1393) reduced the ischemia-induced increase in Ca
2+
. We show that myelinic
N
-methyl-
D
-aspartate (NMDA) receptors are activated by the two conventional coagonists glutamate and glycine, which are released by specific transporters under conditions of cellular Na
+
loading and depolarization in injured white matter. This new technique facilitates detailed studies of living myelin, a vital component of the mammalian CNS.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>17603496</pmid><doi>10.1038/nm1568</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1078-8956 |
| ispartof | Nature medicine, 2007-07, Vol.13 (7), p.874-879 |
| issn | 1078-8956 1546-170X |
| language | eng |
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| source | MEDLINE; Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | Animals Biomedical and Life Sciences Biomedicine Calcium - metabolism Calcium Signaling Cancer Research Central nervous system Central Nervous System - cytology Central Nervous System - metabolism Fluorescence Fluorescent Dyes Infectious Diseases Mammals Metabolic Diseases Microscopy Molecular Medicine Myelin Sheath - metabolism Nervous system Neurons - cytology Neurosciences Rats Rats, Long-Evans technical-report Time Factors |
| title | Real-time measurement of free Ca2+ changes in CNS myelin by two-photon microscopy |
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