Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells
In this study, we investigated whether nucleoplasmic free Ca2+ in aortic vascular smooth muscle cells (VSMCs) might be independently regulated from cytosolic free Ca2+. Understanding mechanisms and pathways responsible for this regulation is especially relevant given the role of a numerous intranucl...
Published in: | Canadian Journal of Physiology and Pharmacology |
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Online Access: | http://hdl.handle.net/1993/2915 https://doi.org/10.1139/y03-005 |
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ftunivmanitoba:oai:mspace.lib.umanitoba.ca:1993/2915 2023-06-18T03:41:16+02:00 Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells Abrenica, B Pierce, GN Gilchrist, JSC 2003-09-30 931128 bytes application/pdf http://hdl.handle.net/1993/2915 https://doi.org/10.1139/y03-005 eng eng CAN J EARTH SCI, SEP 2003, vol. 40, no. 9, p.1259 to 1278. http://hdl.handle.net/1993/2915 http://dx.doi.org/10.1139/y03-005 No part of the NRC Research Press electronic journals may be reproduced, stored, or transmitted in any form or by any means, without the written permission of the publisher, except as stated below. Under the Canadian Copyright Act, individuals may download or print single copies of articles for personal research or study. Any person may reproduce short excerpts from articles in the journals for any purpose that respects the moral rights of authors, provided that the source is fully acknowledged. As a courtesy, the consent of authors of such material should be obtained directly from the author. Authorization to reproduce items for other than personal research or study, as stated above, may be obtained via Access © upon payment of the copyright fee of $10.00 per copy. NRC Research Press also extends certain additional rights to authors. The above rights do not extend to copying or reproduction for general distribution, for advertising or promotional purposes, for creating new collective works, or for resale. For such copying or reproduction, arrangements must be made with NRC Research Press. open access LAURENTIDE ICE-SHEET ABRUPT CLIMATE-CHANGE C-14 YR BP 3-OUTLET CONTROL LARGE BEACHES VOLUME BASIN SEDIMENTATION SASKATCHEWAN BATHYMETRY journal article 2003 ftunivmanitoba https://doi.org/10.1139/y03-005 2023-06-04T17:45:18Z In this study, we investigated whether nucleoplasmic free Ca2+ in aortic vascular smooth muscle cells (VSMCs) might be independently regulated from cytosolic free Ca2+. Understanding mechanisms and pathways responsible for this regulation is especially relevant given the role of a numerous intranuclear Ca2+-sensitive proteins in transcriptional regulation, apoptosis and cell division. The question of an independent regulatory mechanism remains largely unsettled because the previous use of intensitometric fluorophores (e.g., Fluo-3) has been criticized on technical grounds. To circumvent the potential problem of fluorescence artifact, we utilized confocal laser scanning microscopy to image intracellular Ca2+ movements with the ratiometric fluorophore Indo-1. In cultured rabbit VSMCs, we found sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA) pumps and ryanodine receptor (RyR) Ca2+ channel proteins to be discretely arranged within a perinuclear locus, as determined by fluorescent staining patterns of BODIPY(R) FL thapsigargin and BODIPY(R) FL-X Ry. When intracellular Ca2+ stores were mobilized by addition of thapsigargin (5 muM) and activatory concentrations of ryanodine (1 muM), Indo-1 ratiometric signals were largely restricted to the nucleoplasm. Cytosolic signals, by comparison, were relatively small and even then its spatial distribution was largely perinuclear rather homogeneous. These observations indicate perinuclear RyR and SERCA proteins are intimately involved in regulating VSMC nucleoplasmic Ca2+ concentrations. We also observed a similar pattern of largely nucleoplasmic Ca2+ mobilization upon exposure of cells to the immunosuppressant drug FK506 (tacrolimus), which binds to the RyR-associated immunophillin-binding proteins FKBP12 and FKBP12.6. However, initial FK506-induced nucleoplasmic Ca2+ mobilization was followed by marked reduction of Indo-1 signal intensity close to pretreatment levels. This suggested FK506 exerts both activatory and inhibitory effects upon RyR channels. The latter was reinforced ... Article in Journal/Newspaper Ice Sheet MSpace at the University of Manitoba Canadian Journal of Physiology and Pharmacology 81 3 301 310 |
institution |
Open Polar |
collection |
MSpace at the University of Manitoba |
op_collection_id |
ftunivmanitoba |
language |
English |
topic |
LAURENTIDE ICE-SHEET ABRUPT CLIMATE-CHANGE C-14 YR BP 3-OUTLET CONTROL LARGE BEACHES VOLUME BASIN SEDIMENTATION SASKATCHEWAN BATHYMETRY |
spellingShingle |
LAURENTIDE ICE-SHEET ABRUPT CLIMATE-CHANGE C-14 YR BP 3-OUTLET CONTROL LARGE BEACHES VOLUME BASIN SEDIMENTATION SASKATCHEWAN BATHYMETRY Abrenica, B Pierce, GN Gilchrist, JSC Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
topic_facet |
LAURENTIDE ICE-SHEET ABRUPT CLIMATE-CHANGE C-14 YR BP 3-OUTLET CONTROL LARGE BEACHES VOLUME BASIN SEDIMENTATION SASKATCHEWAN BATHYMETRY |
description |
In this study, we investigated whether nucleoplasmic free Ca2+ in aortic vascular smooth muscle cells (VSMCs) might be independently regulated from cytosolic free Ca2+. Understanding mechanisms and pathways responsible for this regulation is especially relevant given the role of a numerous intranuclear Ca2+-sensitive proteins in transcriptional regulation, apoptosis and cell division. The question of an independent regulatory mechanism remains largely unsettled because the previous use of intensitometric fluorophores (e.g., Fluo-3) has been criticized on technical grounds. To circumvent the potential problem of fluorescence artifact, we utilized confocal laser scanning microscopy to image intracellular Ca2+ movements with the ratiometric fluorophore Indo-1. In cultured rabbit VSMCs, we found sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA) pumps and ryanodine receptor (RyR) Ca2+ channel proteins to be discretely arranged within a perinuclear locus, as determined by fluorescent staining patterns of BODIPY(R) FL thapsigargin and BODIPY(R) FL-X Ry. When intracellular Ca2+ stores were mobilized by addition of thapsigargin (5 muM) and activatory concentrations of ryanodine (1 muM), Indo-1 ratiometric signals were largely restricted to the nucleoplasm. Cytosolic signals, by comparison, were relatively small and even then its spatial distribution was largely perinuclear rather homogeneous. These observations indicate perinuclear RyR and SERCA proteins are intimately involved in regulating VSMC nucleoplasmic Ca2+ concentrations. We also observed a similar pattern of largely nucleoplasmic Ca2+ mobilization upon exposure of cells to the immunosuppressant drug FK506 (tacrolimus), which binds to the RyR-associated immunophillin-binding proteins FKBP12 and FKBP12.6. However, initial FK506-induced nucleoplasmic Ca2+ mobilization was followed by marked reduction of Indo-1 signal intensity close to pretreatment levels. This suggested FK506 exerts both activatory and inhibitory effects upon RyR channels. The latter was reinforced ... |
format |
Article in Journal/Newspaper |
author |
Abrenica, B Pierce, GN Gilchrist, JSC |
author_facet |
Abrenica, B Pierce, GN Gilchrist, JSC |
author_sort |
Abrenica, B |
title |
Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
title_short |
Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
title_full |
Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
title_fullStr |
Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
title_full_unstemmed |
Nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
title_sort |
nucleoplasmic calcium regulation in rabbit aortic vascular smooth muscle cells |
publishDate |
2003 |
url |
http://hdl.handle.net/1993/2915 https://doi.org/10.1139/y03-005 |
genre |
Ice Sheet |
genre_facet |
Ice Sheet |
op_relation |
CAN J EARTH SCI, SEP 2003, vol. 40, no. 9, p.1259 to 1278. http://hdl.handle.net/1993/2915 http://dx.doi.org/10.1139/y03-005 |
op_rights |
No part of the NRC Research Press electronic journals may be reproduced, stored, or transmitted in any form or by any means, without the written permission of the publisher, except as stated below. Under the Canadian Copyright Act, individuals may download or print single copies of articles for personal research or study. Any person may reproduce short excerpts from articles in the journals for any purpose that respects the moral rights of authors, provided that the source is fully acknowledged. As a courtesy, the consent of authors of such material should be obtained directly from the author. Authorization to reproduce items for other than personal research or study, as stated above, may be obtained via Access © upon payment of the copyright fee of $10.00 per copy. NRC Research Press also extends certain additional rights to authors. The above rights do not extend to copying or reproduction for general distribution, for advertising or promotional purposes, for creating new collective works, or for resale. For such copying or reproduction, arrangements must be made with NRC Research Press. open access |
op_doi |
https://doi.org/10.1139/y03-005 |
container_title |
Canadian Journal of Physiology and Pharmacology |
container_volume |
81 |
container_issue |
3 |
container_start_page |
301 |
op_container_end_page |
310 |
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1769006754405810176 |