Diffusional Mobility of Parvalbumin in Spiny Dendrites of Cerebellar Purkinje Neurons Quantified by Fluorescence Recovery after Photobleaching
Ca 2+-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca 2+ dynamics. Especially in thin extensions of nerve cells, Ca 2+ binding and buffered diffusion of Ca 2+ by CaBPs is assumed to effectively control the spatio-temporal extend of Ca 2+ signals. However, no quantita...
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| Veröffentlicht in: | Biophysical journal 2003-04, Vol.84 (4), p.2599-2608 |
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| creator | Schmidt, Hartmut Brown, Edward B. Schwaller, Beat Eilers, Jens |
| description | Ca
2+-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca
2+ dynamics. Especially in thin extensions of nerve cells, Ca
2+ binding and buffered diffusion of Ca
2+ by CaBPs is assumed to effectively control the spatio-temporal extend of Ca
2+ signals. However, no quantitative data about the mobility of specific CaBPs in the neuronal cytosol are available. We quantified the diffusion of the endogenous CaPB parvalbumin (PV) in spiny dendrites of cerebellar Purkinje neurons with two-photon fluorescence recovery after photobleaching. Fluorescently labeled PV diffused readily between spines and dendrites with a median time constant of 49
ms (37–61
ms, interquartile range). Based on published data on spine geometry, this value corresponds to an apparent diffusion coefficient of 43
μm
2
s
−1 (34–56
μm
2
s
−1). The absence of large or immobile binding partners for PV was confirmed in PV null-mutant mice. Our data validate the common but so far unproven assumption that PV is highly mobile in neurons and will facilitate simulations of neuronal Ca
2+ buffering. Our experimental approach represents a versatile tool for quantifying the mobility of proteins in neuronal dendrites. |
| doi_str_mv | 10.1016/S0006-3495(03)75065-6 |
| format | Article |
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2+-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca
2+ dynamics. Especially in thin extensions of nerve cells, Ca
2+ binding and buffered diffusion of Ca
2+ by CaBPs is assumed to effectively control the spatio-temporal extend of Ca
2+ signals. However, no quantitative data about the mobility of specific CaBPs in the neuronal cytosol are available. We quantified the diffusion of the endogenous CaPB parvalbumin (PV) in spiny dendrites of cerebellar Purkinje neurons with two-photon fluorescence recovery after photobleaching. Fluorescently labeled PV diffused readily between spines and dendrites with a median time constant of 49
ms (37–61
ms, interquartile range). Based on published data on spine geometry, this value corresponds to an apparent diffusion coefficient of 43
μm
2
s
−1 (34–56
μm
2
s
−1). The absence of large or immobile binding partners for PV was confirmed in PV null-mutant mice. Our data validate the common but so far unproven assumption that PV is highly mobile in neurons and will facilitate simulations of neuronal Ca
2+ buffering. Our experimental approach represents a versatile tool for quantifying the mobility of proteins in neuronal dendrites.</description><identifier>ISSN: 0006-3495</identifier><identifier>EISSN: 1542-0086</identifier><identifier>DOI: 10.1016/S0006-3495(03)75065-6</identifier><identifier>PMID: 12668468</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Cerebellum - metabolism ; Culture Techniques ; Dendrites - chemistry ; Dendrites - metabolism ; Diffusion ; Fluorescence Recovery After Photobleaching - methods ; Mice ; Parvalbumins - chemistry ; Parvalbumins - deficiency ; Parvalbumins - metabolism ; Purkinje Cells - chemistry ; Purkinje Cells - metabolism ; Rats ; Reproducibility of Results ; Sensitivity and Specificity ; Spectroscopy, Imaging, Other Techniques</subject><ispartof>Biophysical journal, 2003-04, Vol.84 (4), p.2599-2608</ispartof><rights>2003 The Biophysical Society</rights><rights>Copyright © 2003, Biophysical Society 2003</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c612t-acab97ae77c4eccf6b664b3d7bd374c212d5f39f869405faea478e17674d3db13</citedby><cites>FETCH-LOGICAL-c612t-acab97ae77c4eccf6b664b3d7bd374c212d5f39f869405faea478e17674d3db13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1302826/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0006349503750656$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,3537,27901,27902,53766,53768,65534</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12668468$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Schmidt, Hartmut</creatorcontrib><creatorcontrib>Brown, Edward B.</creatorcontrib><creatorcontrib>Schwaller, Beat</creatorcontrib><creatorcontrib>Eilers, Jens</creatorcontrib><title>Diffusional Mobility of Parvalbumin in Spiny Dendrites of Cerebellar Purkinje Neurons Quantified by Fluorescence Recovery after Photobleaching</title><title>Biophysical journal</title><addtitle>Biophys J</addtitle><description>Ca
2+-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca
2+ dynamics. Especially in thin extensions of nerve cells, Ca
2+ binding and buffered diffusion of Ca
2+ by CaBPs is assumed to effectively control the spatio-temporal extend of Ca
2+ signals. However, no quantitative data about the mobility of specific CaBPs in the neuronal cytosol are available. We quantified the diffusion of the endogenous CaPB parvalbumin (PV) in spiny dendrites of cerebellar Purkinje neurons with two-photon fluorescence recovery after photobleaching. Fluorescently labeled PV diffused readily between spines and dendrites with a median time constant of 49
ms (37–61
ms, interquartile range). Based on published data on spine geometry, this value corresponds to an apparent diffusion coefficient of 43
μm
2
s
−1 (34–56
μm
2
s
−1). The absence of large or immobile binding partners for PV was confirmed in PV null-mutant mice. Our data validate the common but so far unproven assumption that PV is highly mobile in neurons and will facilitate simulations of neuronal Ca
2+ buffering. Our experimental approach represents a versatile tool for quantifying the mobility of proteins in neuronal dendrites.</description><subject>Animals</subject><subject>Cerebellum - metabolism</subject><subject>Culture Techniques</subject><subject>Dendrites - chemistry</subject><subject>Dendrites - metabolism</subject><subject>Diffusion</subject><subject>Fluorescence Recovery After Photobleaching - methods</subject><subject>Mice</subject><subject>Parvalbumins - chemistry</subject><subject>Parvalbumins - deficiency</subject><subject>Parvalbumins - metabolism</subject><subject>Purkinje Cells - chemistry</subject><subject>Purkinje Cells - metabolism</subject><subject>Rats</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Spectroscopy, Imaging, Other Techniques</subject><issn>0006-3495</issn><issn>1542-0086</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9u1DAQhy0EokvhEUA-ITgE_C92cqFCWwpIBQqFs2U7465L1l7sZKW8BM9MtrsqcKpkyQd_M_7NfAg9peQVJVS-viSEyIqLtn5B-EtVE1lX8h5a0FqwipBG3keLW-QIPSrlmhDKakIfoiPKpGyEbBbo92nwfiwhRdPjT8mGPgwTTh5fmLw1vR3XIeL5XG5CnPApxC6HAcqOWEIGC31vMr4Y888QrwF_hjGnWPDX0cQh-AAdthM-68eUoTiIDvA3cGkLecLGDzCXrtKQbA_GrUK8eoweeNMXeHK4j9GPs3fflx-q8y_vPy7fnldOUjZUxhnbKgNKOQHOeWmlFJZ3ynZcCcco62rPW9_IVpDaGzBCNUCVVKLjnaX8GL3Z992Mdg3dnGzIptebHNYmTzqZoP9_iWGlr9JWU05Yw-Tc4PmhQU6_RiiDXod5wHkbEdJYtOJUqFryO0HaMiJbymaw3oMup1Iy-Ns0lOidcn2jXO98asL1jXK9S_Ls31H-Vh0cz8DJHoB5odsAWRcXdiq6kMENukvhji_-AHiNwGY</recordid><startdate>20030401</startdate><enddate>20030401</enddate><creator>Schmidt, Hartmut</creator><creator>Brown, Edward B.</creator><creator>Schwaller, Beat</creator><creator>Eilers, Jens</creator><general>Elsevier Inc</general><general>Biophysical Society</general><scope>6I.</scope><scope>AAFTH</scope><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>7TK</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20030401</creationdate><title>Diffusional Mobility of Parvalbumin in Spiny Dendrites of Cerebellar Purkinje Neurons Quantified by Fluorescence Recovery after Photobleaching</title><author>Schmidt, Hartmut ; Brown, Edward B. ; Schwaller, Beat ; Eilers, Jens</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c612t-acab97ae77c4eccf6b664b3d7bd374c212d5f39f869405faea478e17674d3db13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Cerebellum - metabolism</topic><topic>Culture Techniques</topic><topic>Dendrites - chemistry</topic><topic>Dendrites - metabolism</topic><topic>Diffusion</topic><topic>Fluorescence Recovery After Photobleaching - methods</topic><topic>Mice</topic><topic>Parvalbumins - chemistry</topic><topic>Parvalbumins - deficiency</topic><topic>Parvalbumins - metabolism</topic><topic>Purkinje Cells - chemistry</topic><topic>Purkinje Cells - metabolism</topic><topic>Rats</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Spectroscopy, Imaging, Other Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Schmidt, Hartmut</creatorcontrib><creatorcontrib>Brown, Edward B.</creatorcontrib><creatorcontrib>Schwaller, Beat</creatorcontrib><creatorcontrib>Eilers, Jens</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biophysical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Schmidt, Hartmut</au><au>Brown, Edward B.</au><au>Schwaller, Beat</au><au>Eilers, Jens</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Diffusional Mobility of Parvalbumin in Spiny Dendrites of Cerebellar Purkinje Neurons Quantified by Fluorescence Recovery after Photobleaching</atitle><jtitle>Biophysical journal</jtitle><addtitle>Biophys J</addtitle><date>2003-04-01</date><risdate>2003</risdate><volume>84</volume><issue>4</issue><spage>2599</spage><epage>2608</epage><pages>2599-2608</pages><issn>0006-3495</issn><eissn>1542-0086</eissn><abstract>Ca
2+-binding proteins (CaBPs) represent key factors for the modulation of cellular Ca
2+ dynamics. Especially in thin extensions of nerve cells, Ca
2+ binding and buffered diffusion of Ca
2+ by CaBPs is assumed to effectively control the spatio-temporal extend of Ca
2+ signals. However, no quantitative data about the mobility of specific CaBPs in the neuronal cytosol are available. We quantified the diffusion of the endogenous CaPB parvalbumin (PV) in spiny dendrites of cerebellar Purkinje neurons with two-photon fluorescence recovery after photobleaching. Fluorescently labeled PV diffused readily between spines and dendrites with a median time constant of 49
ms (37–61
ms, interquartile range). Based on published data on spine geometry, this value corresponds to an apparent diffusion coefficient of 43
μm
2
s
−1 (34–56
μm
2
s
−1). The absence of large or immobile binding partners for PV was confirmed in PV null-mutant mice. Our data validate the common but so far unproven assumption that PV is highly mobile in neurons and will facilitate simulations of neuronal Ca
2+ buffering. Our experimental approach represents a versatile tool for quantifying the mobility of proteins in neuronal dendrites.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>12668468</pmid><doi>10.1016/S0006-3495(03)75065-6</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Biophysical journal, 2003-04, Vol.84 (4), p.2599-2608 |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete; Cell Press Free Archives; EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Animals Cerebellum - metabolism Culture Techniques Dendrites - chemistry Dendrites - metabolism Diffusion Fluorescence Recovery After Photobleaching - methods Mice Parvalbumins - chemistry Parvalbumins - deficiency Parvalbumins - metabolism Purkinje Cells - chemistry Purkinje Cells - metabolism Rats Reproducibility of Results Sensitivity and Specificity Spectroscopy, Imaging, Other Techniques |
| title | Diffusional Mobility of Parvalbumin in Spiny Dendrites of Cerebellar Purkinje Neurons Quantified by Fluorescence Recovery after Photobleaching |
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