Evidence for a Specific Uptake and Retention Mechanism for 25-Hydroxyvitamin D (25OHD) in Skeletal Muscle Cells
Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2...
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| Veröffentlicht in: | Endocrinology (Philadelphia) 2013-09, Vol.154 (9), p.3022-3030 |
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| creator | Abboud, M Puglisi, D. A Davies, B. N Rybchyn, M Whitehead, N. P Brock, K. E Cole, L Gordon-Thomson, C Fraser, D. R Mason, R. S |
| description | Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2 murine muscle cells to take up and release 25OHD, in comparison with other cell types and the involvement of the membrane protein megalin in these mechanisms. When C2 cells are differentiated into myotubes, the time-dependent uptake of labeled 25OHD is 2–3 times higher than in undifferentiated myoblasts or nonmuscle osteoblastic MG63 cells (P < .001). During in vitro release experiments (after 25OHD uptake), myotubes released only 32% ± 6% stored 25OHD after 4 hours, whereas this figure was 60% ± 2% for osteoblasts (P < .01). Using immunofluorescence, C2 myotubes and primary rat muscle fibers were, for the first time, shown to express megalin and cubilin, endocytotic receptors for the vitamin D binding protein (DBP), which binds nearly all 25OHD in the blood. DBP has a high affinity for actin in skeletal muscle. A time-dependent uptake of Alexafluor-488-labeled DBP into mature muscle cells was observed by confocal microscopy. Incubation of C2 myotubes (for 24 hours) with receptor-associated protein, a megalin inhibitor, led to a 40% decrease in 25OHD uptake (P < .01). These data support the proposal that 25OHD, after uptake into mature muscle cells, is held there by DBP, which has been internalized via membrane megalin and is retained by binding to actin. |
| doi_str_mv | 10.1210/en.2012-2245 |
| format | Article |
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A ; Davies, B. N ; Rybchyn, M ; Whitehead, N. P ; Brock, K. E ; Cole, L ; Gordon-Thomson, C ; Fraser, D. R ; Mason, R. S</creator><creatorcontrib>Abboud, M ; Puglisi, D. A ; Davies, B. N ; Rybchyn, M ; Whitehead, N. P ; Brock, K. E ; Cole, L ; Gordon-Thomson, C ; Fraser, D. R ; Mason, R. S</creatorcontrib><description>Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2 murine muscle cells to take up and release 25OHD, in comparison with other cell types and the involvement of the membrane protein megalin in these mechanisms. When C2 cells are differentiated into myotubes, the time-dependent uptake of labeled 25OHD is 2–3 times higher than in undifferentiated myoblasts or nonmuscle osteoblastic MG63 cells (P < .001). During in vitro release experiments (after 25OHD uptake), myotubes released only 32% ± 6% stored 25OHD after 4 hours, whereas this figure was 60% ± 2% for osteoblasts (P < .01). Using immunofluorescence, C2 myotubes and primary rat muscle fibers were, for the first time, shown to express megalin and cubilin, endocytotic receptors for the vitamin D binding protein (DBP), which binds nearly all 25OHD in the blood. DBP has a high affinity for actin in skeletal muscle. A time-dependent uptake of Alexafluor-488-labeled DBP into mature muscle cells was observed by confocal microscopy. Incubation of C2 myotubes (for 24 hours) with receptor-associated protein, a megalin inhibitor, led to a 40% decrease in 25OHD uptake (P < .01). These data support the proposal that 25OHD, after uptake into mature muscle cells, is held there by DBP, which has been internalized via membrane megalin and is retained by binding to actin.</description><identifier>ISSN: 0013-7227</identifier><identifier>EISSN: 1945-7170</identifier><identifier>DOI: 10.1210/en.2012-2245</identifier><identifier>PMID: 23825120</identifier><identifier>CODEN: ENDOAO</identifier><language>eng</language><publisher>Chevy Chase, MD: Endocrine Society</publisher><subject>25-Hydroxyvitamin D ; Actin ; Animals ; Biological and medical sciences ; Blood ; Calcifediol - blood ; Calcifediol - metabolism ; Cell Differentiation ; Cell Line ; Cells, Cultured ; Confocal microscopy ; Endocytosis ; Extracellular Fluid - metabolism ; Female ; Fundamental and applied biological sciences. Psychology ; Immunofluorescence ; Low Density Lipoprotein Receptor-Related Protein-2 - antagonists & inhibitors ; Low Density Lipoprotein Receptor-Related Protein-2 - metabolism ; Male ; Membranes ; Mice ; Mice, Inbred BALB C ; Motor Activity ; Muscle Fibers, Skeletal - cytology ; Muscle Fibers, Skeletal - metabolism ; Muscle, Skeletal - cytology ; Muscle, Skeletal - metabolism ; Muscles ; Musculoskeletal system ; Myoblasts ; Myoblasts, Skeletal - cytology ; Myoblasts, Skeletal - metabolism ; Myotubes ; Osteoblasts ; Osteoblasts - cytology ; Osteoblasts - metabolism ; Proteins ; Random Allocation ; Receptors ; Receptors, Cell Surface - metabolism ; Skeletal muscle ; Striated muscle. Tendons ; Time dependence ; Vertebrates: endocrinology ; Vertebrates: osteoarticular system, musculoskeletal system ; Vitamin D ; Vitamin D-Binding Protein - metabolism</subject><ispartof>Endocrinology (Philadelphia), 2013-09, Vol.154 (9), p.3022-3030</ispartof><rights>Copyright © 2013 by The Endocrine Society</rights><rights>Copyright © 2013 by The Endocrine Society 2013</rights><rights>2014 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c529t-904c5b61432d7d666cbd3195a01ca6bc1dc4ed33fc379e1f85a6c6ffd2b57ff23</citedby><cites>FETCH-LOGICAL-c529t-904c5b61432d7d666cbd3195a01ca6bc1dc4ed33fc379e1f85a6c6ffd2b57ff23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=27663264$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23825120$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Abboud, M</creatorcontrib><creatorcontrib>Puglisi, D. A</creatorcontrib><creatorcontrib>Davies, B. N</creatorcontrib><creatorcontrib>Rybchyn, M</creatorcontrib><creatorcontrib>Whitehead, N. P</creatorcontrib><creatorcontrib>Brock, K. E</creatorcontrib><creatorcontrib>Cole, L</creatorcontrib><creatorcontrib>Gordon-Thomson, C</creatorcontrib><creatorcontrib>Fraser, D. R</creatorcontrib><creatorcontrib>Mason, R. S</creatorcontrib><title>Evidence for a Specific Uptake and Retention Mechanism for 25-Hydroxyvitamin D (25OHD) in Skeletal Muscle Cells</title><title>Endocrinology (Philadelphia)</title><addtitle>Endocrinology</addtitle><description>Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2 murine muscle cells to take up and release 25OHD, in comparison with other cell types and the involvement of the membrane protein megalin in these mechanisms. When C2 cells are differentiated into myotubes, the time-dependent uptake of labeled 25OHD is 2–3 times higher than in undifferentiated myoblasts or nonmuscle osteoblastic MG63 cells (P < .001). During in vitro release experiments (after 25OHD uptake), myotubes released only 32% ± 6% stored 25OHD after 4 hours, whereas this figure was 60% ± 2% for osteoblasts (P < .01). Using immunofluorescence, C2 myotubes and primary rat muscle fibers were, for the first time, shown to express megalin and cubilin, endocytotic receptors for the vitamin D binding protein (DBP), which binds nearly all 25OHD in the blood. DBP has a high affinity for actin in skeletal muscle. A time-dependent uptake of Alexafluor-488-labeled DBP into mature muscle cells was observed by confocal microscopy. Incubation of C2 myotubes (for 24 hours) with receptor-associated protein, a megalin inhibitor, led to a 40% decrease in 25OHD uptake (P < .01). These data support the proposal that 25OHD, after uptake into mature muscle cells, is held there by DBP, which has been internalized via membrane megalin and is retained by binding to actin.</description><subject>25-Hydroxyvitamin D</subject><subject>Actin</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Blood</subject><subject>Calcifediol - blood</subject><subject>Calcifediol - metabolism</subject><subject>Cell Differentiation</subject><subject>Cell Line</subject><subject>Cells, Cultured</subject><subject>Confocal microscopy</subject><subject>Endocytosis</subject><subject>Extracellular Fluid - metabolism</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Immunofluorescence</subject><subject>Low Density Lipoprotein Receptor-Related Protein-2 - antagonists & inhibitors</subject><subject>Low Density Lipoprotein Receptor-Related Protein-2 - metabolism</subject><subject>Male</subject><subject>Membranes</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Motor Activity</subject><subject>Muscle Fibers, Skeletal - cytology</subject><subject>Muscle Fibers, Skeletal - metabolism</subject><subject>Muscle, Skeletal - cytology</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Muscles</subject><subject>Musculoskeletal system</subject><subject>Myoblasts</subject><subject>Myoblasts, Skeletal - cytology</subject><subject>Myoblasts, Skeletal - metabolism</subject><subject>Myotubes</subject><subject>Osteoblasts</subject><subject>Osteoblasts - cytology</subject><subject>Osteoblasts - metabolism</subject><subject>Proteins</subject><subject>Random Allocation</subject><subject>Receptors</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Skeletal muscle</subject><subject>Striated muscle. Tendons</subject><subject>Time dependence</subject><subject>Vertebrates: endocrinology</subject><subject>Vertebrates: osteoarticular system, musculoskeletal system</subject><subject>Vitamin D</subject><subject>Vitamin D-Binding Protein - metabolism</subject><issn>0013-7227</issn><issn>1945-7170</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10d9rFDEQB_BFFHutvvksAREruDWZ_NjbR7lWT2gpWPu8ZJMJpt1Ntslu8f777nmnBdGnMPBhZjLfonjF6AkDRj9iOAHKoAQQ8kmxYLWQZcUq-rRYUMp4WQFUB8VhzjdzKYTgz4sD4EuQDOiiiGf33mIwSFxMRJOrAY133pDrYdS3SHSw5BuOGEYfA7lA80MHn_tfGmS53tgUf27u_ah7H8gpOQZ5uT59T-bi6hY7HHVHLqZsOiQr7Lr8onjmdJfx5f49Kq4_n31frcvzyy9fV5_OSyOhHsuaCiNbxQQHW1mllGktZ7XUlBmtWsOsEWg5d4ZXNTK3lFoZ5ZyFVlbOAT8qjnd9hxTvJsxj0_ts5g10wDjlhglYQgWyrmf65i96E6cU5u0azjhVIFjFZ_Vhp0yKOSd0zZB8r9OmYbTZBtFgaLZBNNsgZv5633Rqe7R_8O_Lz-DtHuhsdOeSDsbnR1cpxUGJ2b3buTgN_xtZ7kfyncRgo0k-4JAw58ff_HPRB-1MqzU</recordid><startdate>20130901</startdate><enddate>20130901</enddate><creator>Abboud, M</creator><creator>Puglisi, D. A</creator><creator>Davies, B. N</creator><creator>Rybchyn, M</creator><creator>Whitehead, N. P</creator><creator>Brock, K. E</creator><creator>Cole, L</creator><creator>Gordon-Thomson, C</creator><creator>Fraser, D. R</creator><creator>Mason, R. S</creator><general>Endocrine Society</general><general>Oxford University Press</general><scope>IQODW</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>7QG</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7TM</scope><scope>7TO</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20130901</creationdate><title>Evidence for a Specific Uptake and Retention Mechanism for 25-Hydroxyvitamin D (25OHD) in Skeletal Muscle Cells</title><author>Abboud, M ; Puglisi, D. A ; Davies, B. N ; Rybchyn, M ; Whitehead, N. P ; Brock, K. E ; Cole, L ; Gordon-Thomson, C ; Fraser, D. R ; Mason, R. S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c529t-904c5b61432d7d666cbd3195a01ca6bc1dc4ed33fc379e1f85a6c6ffd2b57ff23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>25-Hydroxyvitamin D</topic><topic>Actin</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Blood</topic><topic>Calcifediol - blood</topic><topic>Calcifediol - metabolism</topic><topic>Cell Differentiation</topic><topic>Cell Line</topic><topic>Cells, Cultured</topic><topic>Confocal microscopy</topic><topic>Endocytosis</topic><topic>Extracellular Fluid - metabolism</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Immunofluorescence</topic><topic>Low Density Lipoprotein Receptor-Related Protein-2 - antagonists & inhibitors</topic><topic>Low Density Lipoprotein Receptor-Related Protein-2 - metabolism</topic><topic>Male</topic><topic>Membranes</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Motor Activity</topic><topic>Muscle Fibers, Skeletal - cytology</topic><topic>Muscle Fibers, Skeletal - metabolism</topic><topic>Muscle, Skeletal - cytology</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Muscles</topic><topic>Musculoskeletal system</topic><topic>Myoblasts</topic><topic>Myoblasts, Skeletal - cytology</topic><topic>Myoblasts, Skeletal - metabolism</topic><topic>Myotubes</topic><topic>Osteoblasts</topic><topic>Osteoblasts - cytology</topic><topic>Osteoblasts - metabolism</topic><topic>Proteins</topic><topic>Random Allocation</topic><topic>Receptors</topic><topic>Receptors, Cell Surface - metabolism</topic><topic>Skeletal muscle</topic><topic>Striated muscle. Tendons</topic><topic>Time dependence</topic><topic>Vertebrates: endocrinology</topic><topic>Vertebrates: osteoarticular system, musculoskeletal system</topic><topic>Vitamin D</topic><topic>Vitamin D-Binding Protein - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Abboud, M</creatorcontrib><creatorcontrib>Puglisi, D. A</creatorcontrib><creatorcontrib>Davies, B. N</creatorcontrib><creatorcontrib>Rybchyn, M</creatorcontrib><creatorcontrib>Whitehead, N. P</creatorcontrib><creatorcontrib>Brock, K. E</creatorcontrib><creatorcontrib>Cole, L</creatorcontrib><creatorcontrib>Gordon-Thomson, C</creatorcontrib><creatorcontrib>Fraser, D. R</creatorcontrib><creatorcontrib>Mason, R. S</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Immunology Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Endocrinology (Philadelphia)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Abboud, M</au><au>Puglisi, D. A</au><au>Davies, B. N</au><au>Rybchyn, M</au><au>Whitehead, N. P</au><au>Brock, K. E</au><au>Cole, L</au><au>Gordon-Thomson, C</au><au>Fraser, D. R</au><au>Mason, R. S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evidence for a Specific Uptake and Retention Mechanism for 25-Hydroxyvitamin D (25OHD) in Skeletal Muscle Cells</atitle><jtitle>Endocrinology (Philadelphia)</jtitle><addtitle>Endocrinology</addtitle><date>2013-09-01</date><risdate>2013</risdate><volume>154</volume><issue>9</issue><spage>3022</spage><epage>3030</epage><pages>3022-3030</pages><issn>0013-7227</issn><eissn>1945-7170</eissn><coden>ENDOAO</coden><abstract>Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2 murine muscle cells to take up and release 25OHD, in comparison with other cell types and the involvement of the membrane protein megalin in these mechanisms. When C2 cells are differentiated into myotubes, the time-dependent uptake of labeled 25OHD is 2–3 times higher than in undifferentiated myoblasts or nonmuscle osteoblastic MG63 cells (P < .001). During in vitro release experiments (after 25OHD uptake), myotubes released only 32% ± 6% stored 25OHD after 4 hours, whereas this figure was 60% ± 2% for osteoblasts (P < .01). Using immunofluorescence, C2 myotubes and primary rat muscle fibers were, for the first time, shown to express megalin and cubilin, endocytotic receptors for the vitamin D binding protein (DBP), which binds nearly all 25OHD in the blood. DBP has a high affinity for actin in skeletal muscle. A time-dependent uptake of Alexafluor-488-labeled DBP into mature muscle cells was observed by confocal microscopy. Incubation of C2 myotubes (for 24 hours) with receptor-associated protein, a megalin inhibitor, led to a 40% decrease in 25OHD uptake (P < .01). These data support the proposal that 25OHD, after uptake into mature muscle cells, is held there by DBP, which has been internalized via membrane megalin and is retained by binding to actin.</abstract><cop>Chevy Chase, MD</cop><pub>Endocrine Society</pub><pmid>23825120</pmid><doi>10.1210/en.2012-2245</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Endocrinology (Philadelphia), 2013-09, Vol.154 (9), p.3022-3030 |
| issn | 0013-7227 1945-7170 |
| language | eng |
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| source | Journals@Ovid Ovid Autoload; MEDLINE; Oxford Journals Online; Alma/SFX Local Collection; EZB Electronic Journals Library |
| subjects | 25-Hydroxyvitamin D Actin Animals Biological and medical sciences Blood Calcifediol - blood Calcifediol - metabolism Cell Differentiation Cell Line Cells, Cultured Confocal microscopy Endocytosis Extracellular Fluid - metabolism Female Fundamental and applied biological sciences. Psychology Immunofluorescence Low Density Lipoprotein Receptor-Related Protein-2 - antagonists & inhibitors Low Density Lipoprotein Receptor-Related Protein-2 - metabolism Male Membranes Mice Mice, Inbred BALB C Motor Activity Muscle Fibers, Skeletal - cytology Muscle Fibers, Skeletal - metabolism Muscle, Skeletal - cytology Muscle, Skeletal - metabolism Muscles Musculoskeletal system Myoblasts Myoblasts, Skeletal - cytology Myoblasts, Skeletal - metabolism Myotubes Osteoblasts Osteoblasts - cytology Osteoblasts - metabolism Proteins Random Allocation Receptors Receptors, Cell Surface - metabolism Skeletal muscle Striated muscle. Tendons Time dependence Vertebrates: endocrinology Vertebrates: osteoarticular system, musculoskeletal system Vitamin D Vitamin D-Binding Protein - metabolism |
| title | Evidence for a Specific Uptake and Retention Mechanism for 25-Hydroxyvitamin D (25OHD) in Skeletal Muscle Cells |
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