| Different roles of inositol 1,4,5-trisphosphate receptor subtypes in prostaglandin F(2alpha)-induced calcium oscillations and pacemaking activity of NRK fibroblasts. | |
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MedLine Citation:
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PMID: 20542562 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We investigated the role of inositol 1,4,5-trisphosphate (IP(3))-receptor isoforms in the prostaglandin F(2alpha) (PGF(2alpha))-induced calcium oscillations and pacemaking activity of normal rat kidney (NRK) fibroblasts. Reverse transcription polymerase chain reaction (RT-PCR) studies revealed that NRK fibroblasts express only the IP(3)-receptor subtypes IP(3)R1 and IP(3)R3. Quantitative RT-PCR analysis demonstrated that their expression levels varied as a function of the growth status of NRK cells; NRK cells made quiescent (Q) by serum deprivation expressed significantly higher levels of subtypes 1 and 3 than cells grown to density-arrest (DA). Using Ca(2+)-imaging techniques, we show that the frequency of PGF(2alpha)-induced calcium oscillations in DA-cells is lower than in Q-cells. To study whether these differences in the frequency of calcium oscillations relate to the relative amounts of IP(3)-receptor subtypes expressed by the cells, we knocked down the genes for either IP(3)-receptor subtype by using an shRNA approach. Knockdown of the IP(3)R1 gene significantly decreased the frequency of the PGF(2alpha)-induced calcium oscillations in both Q- and DA-cells. It also reduced the frequency of the repetitive firing of calcium action potentials by DA-cells. In contrast, knockdown of the IP(3)R3 gene caused an increase in the frequency of both processes, suggesting a role for this receptor subtype as an anti-Ca(2+)-oscillatory unit in NRK fibroblasts. Our findings indicate that the reduction in the frequency of PGF(2alpha)-induced calcium oscillations in DA-cells compared with Q-cells results from the reduced expression ratio of IP(3)R1 versus IP(3)R3 receptors in DA-cells. Moreover, these data provide direct evidence that the frequency of IP(3)-dependent calcium oscillations determines the periodicity of action potential firing by density-arrested NRK fibroblasts. |
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Authors:
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W H M Almirza; P H J Peters; W P M van Meerwijk; E J J van Zoelen; A P R Theuvenet |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-06-09 |
Journal Detail:
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Title: Cell calcium Volume: 47 ISSN: 1532-1991 ISO Abbreviation: Cell Calcium Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-07-08 Completed Date: 2010-10-07 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8006226 Medline TA: Cell Calcium Country: Netherlands |
Other Details:
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Languages: eng Pagination: 544-53 Citation Subset: IM |
Copyright Information:
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2010 Elsevier Ltd. All rights reserved. |
Affiliation:
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Department of Cell Biology, Radboud University Nijmegen, The Netherlands. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Calcium / metabolism* Dinoprost / pharmacology* Fibroblasts / metabolism* Gene Knockdown Techniques Inositol 1,4,5-Trisphosphate Receptors / genetics, metabolism, physiology* Kidney / cytology Patch-Clamp Techniques Protein Isoforms / genetics, metabolism, physiology RNA Interference Rats |
| Chemical | |
Reg. No./Substance:
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0/Inositol 1,4,5-Trisphosphate Receptors; 0/Protein Isoforms; 551-11-1/Dinoprost; 7440-70-2/Calcium |
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