Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging
The photoactivatable calcium sensors reported in this paper allow simultaneous highlighting of cellular morphology and recording of calcium activity, which is demonstrated in neuronal cultures, in Drosophila and in zebrafish. Circuit mapping requires knowledge of both structural and functional conne...
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| Veröffentlicht in: | Nature methods 2015-09, Vol.12 (9), p.852-858 |
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| creator | Berlin, Shai Carroll, Elizabeth C Newman, Zachary L Okada, Hitomi O Quinn, Carson M Kallman, Benjamin Rockwell, Nathan C Martin, Shelley S Lagarias, J Clark Isacoff, Ehud Y |
| description | The photoactivatable calcium sensors reported in this paper allow simultaneous highlighting of cellular morphology and recording of calcium activity, which is demonstrated in neuronal cultures, in
Drosophila
and in zebrafish.
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca
2+
indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca
2+
detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP). |
| doi_str_mv | 10.1038/nmeth.3480 |
| format | Article |
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Drosophila
and in zebrafish.
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca
2+
indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca
2+
detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).</description><identifier>ISSN: 1548-7091</identifier><identifier>EISSN: 1548-7105</identifier><identifier>DOI: 10.1038/nmeth.3480</identifier><identifier>PMID: 26167640</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>14 ; 14/19 ; 14/35 ; 631/378/87 ; Animals ; Bioinformatics ; Biological Microscopy ; Biological Techniques ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Calcium - metabolism ; Calcium Signaling - physiology ; Cell Tracking - methods ; Cells, Cultured ; Cellular biology ; Danio rerio ; Drosophila ; Fluorescent proteins ; Fruits ; Larvae ; Life Sciences ; Light ; Luminescent Proteins - genetics ; Luminescent Proteins - metabolism ; Medical research ; Medicine, Experimental ; Methods ; Microscopy, Fluorescence - methods ; Morphology ; Neuroimaging ; Neurons ; Neurons - cytology ; Neurons - physiology ; Optogenetics - methods ; Population density ; Probes ; Protein Engineering - methods ; Proteins ; Proteomics ; Rats ; Sensors ; Zebrafish</subject><ispartof>Nature methods, 2015-09, Vol.12 (9), p.852-858</ispartof><rights>Springer Nature America, Inc. 2015</rights><rights>COPYRIGHT 2015 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Sep 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c608t-22ff3649846067c5a38240c4886abb697f6405d413d6af14fb1ca07c192def613</citedby><cites>FETCH-LOGICAL-c608t-22ff3649846067c5a38240c4886abb697f6405d413d6af14fb1ca07c192def613</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/nmeth.3480$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nmeth.3480$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26167640$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Berlin, Shai</creatorcontrib><creatorcontrib>Carroll, Elizabeth C</creatorcontrib><creatorcontrib>Newman, Zachary L</creatorcontrib><creatorcontrib>Okada, Hitomi O</creatorcontrib><creatorcontrib>Quinn, Carson M</creatorcontrib><creatorcontrib>Kallman, Benjamin</creatorcontrib><creatorcontrib>Rockwell, Nathan C</creatorcontrib><creatorcontrib>Martin, Shelley S</creatorcontrib><creatorcontrib>Lagarias, J Clark</creatorcontrib><creatorcontrib>Isacoff, Ehud Y</creatorcontrib><title>Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging</title><title>Nature methods</title><addtitle>Nat Methods</addtitle><addtitle>Nat Methods</addtitle><description>The photoactivatable calcium sensors reported in this paper allow simultaneous highlighting of cellular morphology and recording of calcium activity, which is demonstrated in neuronal cultures, in
Drosophila
and in zebrafish.
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca
2+
indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca
2+
detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).</description><subject>14</subject><subject>14/19</subject><subject>14/35</subject><subject>631/378/87</subject><subject>Animals</subject><subject>Bioinformatics</subject><subject>Biological Microscopy</subject><subject>Biological Techniques</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>Calcium - metabolism</subject><subject>Calcium Signaling - physiology</subject><subject>Cell Tracking - methods</subject><subject>Cells, Cultured</subject><subject>Cellular biology</subject><subject>Danio rerio</subject><subject>Drosophila</subject><subject>Fluorescent proteins</subject><subject>Fruits</subject><subject>Larvae</subject><subject>Life Sciences</subject><subject>Light</subject><subject>Luminescent Proteins - genetics</subject><subject>Luminescent Proteins - metabolism</subject><subject>Medical research</subject><subject>Medicine, Experimental</subject><subject>Methods</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Morphology</subject><subject>Neuroimaging</subject><subject>Neurons</subject><subject>Neurons - cytology</subject><subject>Neurons - physiology</subject><subject>Optogenetics - methods</subject><subject>Population density</subject><subject>Probes</subject><subject>Protein Engineering - 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Y</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging</atitle><jtitle>Nature methods</jtitle><stitle>Nat Methods</stitle><addtitle>Nat Methods</addtitle><date>2015-09-01</date><risdate>2015</risdate><volume>12</volume><issue>9</issue><spage>852</spage><epage>858</epage><pages>852-858</pages><issn>1548-7091</issn><eissn>1548-7105</eissn><abstract>The photoactivatable calcium sensors reported in this paper allow simultaneous highlighting of cellular morphology and recording of calcium activity, which is demonstrated in neuronal cultures, in
Drosophila
and in zebrafish.
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca
2+
indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca
2+
detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><pmid>26167640</pmid><doi>10.1038/nmeth.3480</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 14 14/19 14/35 631/378/87 Animals Bioinformatics Biological Microscopy Biological Techniques Biomedical and Life Sciences Biomedical Engineering/Biotechnology Calcium - metabolism Calcium Signaling - physiology Cell Tracking - methods Cells, Cultured Cellular biology Danio rerio Drosophila Fluorescent proteins Fruits Larvae Life Sciences Light Luminescent Proteins - genetics Luminescent Proteins - metabolism Medical research Medicine, Experimental Methods Microscopy, Fluorescence - methods Morphology Neuroimaging Neurons Neurons - cytology Neurons - physiology Optogenetics - methods Population density Probes Protein Engineering - methods Proteins Proteomics Rats Sensors Zebrafish |
| title | Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging |
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