Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat
In the present study, the labeling efficacy of tracers Fluoro-ruby (FR), Fluoro-emerald (FE), True Blue (TB), Fluoro-Gold (FG), Diamidino Yellow (DY) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) to retrogradely label the cutaneous afferent neurons in the rat was exami...
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| Veröffentlicht in: | Journal of neuroscience methods 2010-08, Vol.191 (2), p.208-214 |
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| description | In the present study, the labeling efficacy of tracers Fluoro-ruby (FR), Fluoro-emerald (FE), True Blue (TB), Fluoro-Gold (FG), Diamidino Yellow (DY) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) to retrogradely label the cutaneous afferent neurons in the rat was examined.
The proximal stump of the transected sural nerve was exposed for 1 hour either to one of the examined dyes (FR, FE, TB, FG, DY and DiI group) in single labeling experiments, or to mixtures of two dyes (TB-FG, FG-DiI, TB-DY and TB-DiI group) in double labeling experiments (
n
=
5 for each group). After 10 days, dorsal root ganglia (DRGs) L3-S1 were harvested, cut to 20
μm thick longitudinal sections and all labeled neurons were counted.
The average numbers of labeled DRG neurons in FR group (1063
±
158; mean
±
SD) and FE group (1067
±
203) were statistically significantly lower than those in TB group (2831
±
379), FG group (2802
±
134), DY group (2888
±
262) or DiI group (2900
±
278) (
p
<
0.05). In double labeling experiments, the average number of double labeled neurons in TB-DY group (2208
±
207) was statistically significantly lower than those in TB-FG group (2775
±
316), FG-DiI group (2921
±
419), or TB-DiI group (2805
±
179) (
p
<
0.05).
Among examined tracers, TB, FG, DY and DiI, have the highest and similar labeling efficacy for retrograde labeling of cutaneous afferent neurons in the rat. The tracers TB, DiI and FG effectively label the same neuronal population in double labeling, therefore, their combinations are most suitable for double retrograde labeling studies of cutaneous afferent neurons in the rat. |
| doi_str_mv | 10.1016/j.jneumeth.2010.06.021 |
| format | Article |
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The proximal stump of the transected sural nerve was exposed for 1 hour either to one of the examined dyes (FR, FE, TB, FG, DY and DiI group) in single labeling experiments, or to mixtures of two dyes (TB-FG, FG-DiI, TB-DY and TB-DiI group) in double labeling experiments (
n
=
5 for each group). After 10 days, dorsal root ganglia (DRGs) L3-S1 were harvested, cut to 20
μm thick longitudinal sections and all labeled neurons were counted.
The average numbers of labeled DRG neurons in FR group (1063
±
158; mean
±
SD) and FE group (1067
±
203) were statistically significantly lower than those in TB group (2831
±
379), FG group (2802
±
134), DY group (2888
±
262) or DiI group (2900
±
278) (
p
<
0.05). In double labeling experiments, the average number of double labeled neurons in TB-DY group (2208
±
207) was statistically significantly lower than those in TB-FG group (2775
±
316), FG-DiI group (2921
±
419), or TB-DiI group (2805
±
179) (
p
<
0.05).
Among examined tracers, TB, FG, DY and DiI, have the highest and similar labeling efficacy for retrograde labeling of cutaneous afferent neurons in the rat. The tracers TB, DiI and FG effectively label the same neuronal population in double labeling, therefore, their combinations are most suitable for double retrograde labeling studies of cutaneous afferent neurons in the rat.</description><identifier>ISSN: 0165-0270</identifier><identifier>EISSN: 1872-678X</identifier><identifier>DOI: 10.1016/j.jneumeth.2010.06.021</identifier><identifier>PMID: 20600316</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Afferent Pathways - cytology ; Afferent Pathways - physiology ; Animals ; Axonal Transport - physiology ; Cell Count - methods ; Cutaneous afferents ; Dissection - methods ; Dorsal root ganglia ; Dorsal root ganglion ; Fluorescent dyes ; Fluorescent Dyes - pharmacokinetics ; Ganglia, Spinal - cytology ; Ganglia, Spinal - physiology ; Male ; Neuroanatomical Tract-Tracing Techniques - methods ; Neuronal Tract-Tracers - pharmacokinetics ; Neurosurgical Procedures - methods ; Peripheral nervous system ; Rat ; Rats ; Rats, Wistar ; Retrograde labeling ; Sensory axons ; Sensory Receptor Cells - cytology ; Sensory Receptor Cells - physiology ; Staining and Labeling - methods ; Sural nerve ; Sural Nerve - cytology ; Sural Nerve - physiology ; Sural Nerve - surgery</subject><ispartof>Journal of neuroscience methods, 2010-08, Vol.191 (2), p.208-214</ispartof><rights>2010 Elsevier B.V.</rights><rights>Copyright (c) 2010 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c399t-388680262b159b630fd38c021cec94e8820cc4f272cd334c2c9f9ddd14f5fbd03</citedby><cites>FETCH-LOGICAL-c399t-388680262b159b630fd38c021cec94e8820cc4f272cd334c2c9f9ddd14f5fbd03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0165027010003456$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20600316$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Žele, Tilen</creatorcontrib><creatorcontrib>Sketelj, Janez</creatorcontrib><creatorcontrib>Bajrović, Fajko F.</creatorcontrib><title>Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat</title><title>Journal of neuroscience methods</title><addtitle>J Neurosci Methods</addtitle><description>In the present study, the labeling efficacy of tracers Fluoro-ruby (FR), Fluoro-emerald (FE), True Blue (TB), Fluoro-Gold (FG), Diamidino Yellow (DY) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) to retrogradely label the cutaneous afferent neurons in the rat was examined.
The proximal stump of the transected sural nerve was exposed for 1 hour either to one of the examined dyes (FR, FE, TB, FG, DY and DiI group) in single labeling experiments, or to mixtures of two dyes (TB-FG, FG-DiI, TB-DY and TB-DiI group) in double labeling experiments (
n
=
5 for each group). After 10 days, dorsal root ganglia (DRGs) L3-S1 were harvested, cut to 20
μm thick longitudinal sections and all labeled neurons were counted.
The average numbers of labeled DRG neurons in FR group (1063
±
158; mean
±
SD) and FE group (1067
±
203) were statistically significantly lower than those in TB group (2831
±
379), FG group (2802
±
134), DY group (2888
±
262) or DiI group (2900
±
278) (
p
<
0.05). In double labeling experiments, the average number of double labeled neurons in TB-DY group (2208
±
207) was statistically significantly lower than those in TB-FG group (2775
±
316), FG-DiI group (2921
±
419), or TB-DiI group (2805
±
179) (
p
<
0.05).
Among examined tracers, TB, FG, DY and DiI, have the highest and similar labeling efficacy for retrograde labeling of cutaneous afferent neurons in the rat. The tracers TB, DiI and FG effectively label the same neuronal population in double labeling, therefore, their combinations are most suitable for double retrograde labeling studies of cutaneous afferent neurons in the rat.</description><subject>Afferent Pathways - cytology</subject><subject>Afferent Pathways - physiology</subject><subject>Animals</subject><subject>Axonal Transport - physiology</subject><subject>Cell Count - methods</subject><subject>Cutaneous afferents</subject><subject>Dissection - methods</subject><subject>Dorsal root ganglia</subject><subject>Dorsal root ganglion</subject><subject>Fluorescent dyes</subject><subject>Fluorescent Dyes - pharmacokinetics</subject><subject>Ganglia, Spinal - cytology</subject><subject>Ganglia, Spinal - physiology</subject><subject>Male</subject><subject>Neuroanatomical Tract-Tracing Techniques - methods</subject><subject>Neuronal Tract-Tracers - pharmacokinetics</subject><subject>Neurosurgical Procedures - methods</subject><subject>Peripheral nervous system</subject><subject>Rat</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>Retrograde labeling</subject><subject>Sensory axons</subject><subject>Sensory Receptor Cells - cytology</subject><subject>Sensory Receptor Cells - physiology</subject><subject>Staining and Labeling - methods</subject><subject>Sural nerve</subject><subject>Sural Nerve - cytology</subject><subject>Sural Nerve - physiology</subject><subject>Sural Nerve - surgery</subject><issn>0165-0270</issn><issn>1872-678X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUFPGzEQhS3UqgToX0B762nTsb3xem-tEIVKSFyoxM3yjsfgaLMbbG-l_HuchvSakyXrezNv3mPsmsOSA1ff18v1SPOG8utSQPkEtQTBz9iC61bUqtXPn9iigKsaRAvn7CKlNQA0Hagv7FyAApBcLZi59T6gxV01-coP8xQpIY25ytEixVSFsYqU4_QSraNqsD0NYXzZ0zhnO9I0p8p6T3EvKpbiNP4T5Veqos1X7LO3Q6KvH-8l-_Pr9unmvn54vPt98_OhRtl1uZZaKw1CiZ6vul5J8E5qLBchYdeQ1gIQGy9agU7KBgV2vnPO8cavfO9AXrJvh7nbOL3NlLLZhHLIMBwsGt225WTdNifJtmQEAqQopDqQGKeUInmzjWFj485wMPsWzNocWzD7FgwoUzwX4fXHirnfkPsvO8ZegB8HgEokfwNFkzDQiORCJMzGTeHUjndbYJ0J</recordid><startdate>20100830</startdate><enddate>20100830</enddate><creator>Žele, Tilen</creator><creator>Sketelj, Janez</creator><creator>Bajrović, Fajko F.</creator><general>Elsevier B.V</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><scope>7TK</scope></search><sort><creationdate>20100830</creationdate><title>Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat</title><author>Žele, Tilen ; Sketelj, Janez ; Bajrović, Fajko F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c399t-388680262b159b630fd38c021cec94e8820cc4f272cd334c2c9f9ddd14f5fbd03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Afferent Pathways - cytology</topic><topic>Afferent Pathways - physiology</topic><topic>Animals</topic><topic>Axonal Transport - physiology</topic><topic>Cell Count - methods</topic><topic>Cutaneous afferents</topic><topic>Dissection - methods</topic><topic>Dorsal root ganglia</topic><topic>Dorsal root ganglion</topic><topic>Fluorescent dyes</topic><topic>Fluorescent Dyes - pharmacokinetics</topic><topic>Ganglia, Spinal - cytology</topic><topic>Ganglia, Spinal - physiology</topic><topic>Male</topic><topic>Neuroanatomical Tract-Tracing Techniques - methods</topic><topic>Neuronal Tract-Tracers - pharmacokinetics</topic><topic>Neurosurgical Procedures - methods</topic><topic>Peripheral nervous system</topic><topic>Rat</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>Retrograde labeling</topic><topic>Sensory axons</topic><topic>Sensory Receptor Cells - cytology</topic><topic>Sensory Receptor Cells - physiology</topic><topic>Staining and Labeling - methods</topic><topic>Sural nerve</topic><topic>Sural Nerve - cytology</topic><topic>Sural Nerve - physiology</topic><topic>Sural Nerve - surgery</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Žele, Tilen</creatorcontrib><creatorcontrib>Sketelj, Janez</creatorcontrib><creatorcontrib>Bajrović, Fajko 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><collection>Neurosciences Abstracts</collection><jtitle>Journal of neuroscience methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Žele, Tilen</au><au>Sketelj, Janez</au><au>Bajrović, Fajko F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat</atitle><jtitle>Journal of neuroscience methods</jtitle><addtitle>J Neurosci Methods</addtitle><date>2010-08-30</date><risdate>2010</risdate><volume>191</volume><issue>2</issue><spage>208</spage><epage>214</epage><pages>208-214</pages><issn>0165-0270</issn><eissn>1872-678X</eissn><abstract>In the present study, the labeling efficacy of tracers Fluoro-ruby (FR), Fluoro-emerald (FE), True Blue (TB), Fluoro-Gold (FG), Diamidino Yellow (DY) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) to retrogradely label the cutaneous afferent neurons in the rat was examined.
The proximal stump of the transected sural nerve was exposed for 1 hour either to one of the examined dyes (FR, FE, TB, FG, DY and DiI group) in single labeling experiments, or to mixtures of two dyes (TB-FG, FG-DiI, TB-DY and TB-DiI group) in double labeling experiments (
n
=
5 for each group). After 10 days, dorsal root ganglia (DRGs) L3-S1 were harvested, cut to 20
μm thick longitudinal sections and all labeled neurons were counted.
The average numbers of labeled DRG neurons in FR group (1063
±
158; mean
±
SD) and FE group (1067
±
203) were statistically significantly lower than those in TB group (2831
±
379), FG group (2802
±
134), DY group (2888
±
262) or DiI group (2900
±
278) (
p
<
0.05). In double labeling experiments, the average number of double labeled neurons in TB-DY group (2208
±
207) was statistically significantly lower than those in TB-FG group (2775
±
316), FG-DiI group (2921
±
419), or TB-DiI group (2805
±
179) (
p
<
0.05).
Among examined tracers, TB, FG, DY and DiI, have the highest and similar labeling efficacy for retrograde labeling of cutaneous afferent neurons in the rat. The tracers TB, DiI and FG effectively label the same neuronal population in double labeling, therefore, their combinations are most suitable for double retrograde labeling studies of cutaneous afferent neurons in the rat.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>20600316</pmid><doi>10.1016/j.jneumeth.2010.06.021</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Journal of neuroscience methods, 2010-08, Vol.191 (2), p.208-214 |
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| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Afferent Pathways - cytology Afferent Pathways - physiology Animals Axonal Transport - physiology Cell Count - methods Cutaneous afferents Dissection - methods Dorsal root ganglia Dorsal root ganglion Fluorescent dyes Fluorescent Dyes - pharmacokinetics Ganglia, Spinal - cytology Ganglia, Spinal - physiology Male Neuroanatomical Tract-Tracing Techniques - methods Neuronal Tract-Tracers - pharmacokinetics Neurosurgical Procedures - methods Peripheral nervous system Rat Rats Rats, Wistar Retrograde labeling Sensory axons Sensory Receptor Cells - cytology Sensory Receptor Cells - physiology Staining and Labeling - methods Sural nerve Sural Nerve - cytology Sural Nerve - physiology Sural Nerve - surgery |
| title | Efficacy of fluorescent tracers in retrograde labeling of cutaneous afferent neurons in the rat |
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