Growth-dependent modulation of capacitative calcium entry in normal rat kidney fibroblasts |
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| Authors: | MM Dernison WHMA Almirza JMAM Kusters WPM van Meerwijk CCAM Gielen EJJ van Zoelen APR Theuvenet |
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| Institution: | 1. Department of Cell Biology, Radboud University Nijmegen, Heyendaalseweg 135, 6525 AJ Nijmegen, The Netherlands;2. Department of Biophysics, Radboud University Nijmegen, Geert Grooteplein 21, 6525 EZ Nijmegen, The Netherlands;1. Programa de Pós-graduação em Zootecnia, Centro de Educação Superior do Oeste (CEO), Universidade do Estado de Santa Catarina, UDESC, Rua Beloni Trombeta Zanin 680E, 89815-630 Chapecó, Santa Catarina, Brazil;2. Universidade do Contestado, Rua Victor Sopelsa, 3000, 89700-000 Concórdia, Santa Catarina, Brazil;3. Embrapa Suínos e Aves, Rodovia BR-153, Km 110, Distrito de Tamanduá, Caixa Postal: 321, 89715-899 Concórdia, Santa Catarina, Brazil;4. Departamento de Zootecnia, Universidade do Estado de Santa Catarina, Avenida Luiz de Camões, 2090, 88520-000 Lages, Santa Catarina, Brazil;1. University of Groningen, University Medical Center Groningen, Department of Epidemiology, GRIAC Research Institute, Groningen, The Netherlands;2. University of Groningen, University Medical Center Groningen, Department of Pediatrics, Beatrix Children''s Hospital, GRIAC Research Institute, Groningen, The Netherlands;3. Systems and Modeling Unit, Montefiore Institute, University of Liege, Liege, Belgium;4. Bioinformatics and Modeling, GIGA-R, University of Liege, Liege, Belgium;5. School of Social and Community Medicine, University of Bristol, Bristol, United Kingdom;6. University of Groningen, University Medical Center Groningen, Department of Pediatric Pulmonology and Pediatric Allergology, Beatrix Children''s Hospital, GRIAC Research Institute, Groningen, The Netherlands;7. Department of Pediatrics/Respiratory Medicine, Erasmus University Medical Center–Sophia Children''s Hospital, Rotterdam, The Netherlands;8. Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht, The Netherlands;9. Institute for Risk Assessment Sciences, Utrecht University, Utrecht, The Netherlands;10. University of Groningen, University Medical Center Groningen, Department of Pulmonology, GRIAC Research Institute, Groningen, The Netherlands;1. Department of Internal Medicine and Nephrology, University Hospital Giessen and Marburg, Philipps-University Marburg, 35033 Marburg, Germany;2. Department of Cell Biology and Cell Pathology, Philipps-University Marburg, 35037 Marburg, Germany;1. Embrapa Suínos e Aves, BR153 Km 110 – Vila Tamanduá, PO Box 321, 89715-899, Concórdia, SC, Brazil;2. Faculdade de Agronomia e Medicina Veterinária, Curso de Medicina Veterinária, Universidade de Passo Fundo, BR 285, São José, 99052-900, Passo Fundo, RS, Brazil;2. Orthopaedic Biomechanics Laboratories, Michigan State University, East Lansing Michigan 48824;3. Department of Animal and Food Sciences, Texas Tech University, Lubbock, Texas 79409 |
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| Abstract: | Normal rat kidney (NRK) fibroblasts have electrophysiological properties and intracellular calcium dynamics that are dependent upon their growth stage. In the present study we show that this differential behavior coincides with a differential calcium entry that can be either capacitative or non-capacitative. Confluent cells made quiescent by serum deprivation, which have a stable membrane potential near ? 70 mV and do not show spontaneous intracellular calcium oscillations, primarily exhibit the capacitative mechanism for calcium entry, also called store-operated calcium entry (SOCE). When the quiescent cells are grown to density-arrest in the presence of EGF as the sole polypeptide growth factor, these cells characteristically fire spontaneously repetitive calcium action potentials, which propagate throughout the whole monolayer and are accompanied by intracellular calcium transients. These density-arrested cells appear to exhibit in addition to SOCE also receptor-operated calcium entry (ROCE) as a mechanism for calcium entry. Furthermore we show that, in contrast to earlier studies, the employed SOCs and ROCs are permeable for both calcium and strontium ions. We examined the expression of the canonical transient receptor potential channels (Trpcs) that may be involved in SOCE and ROCE. We show that NRK fibroblasts express the genes encoding Trpc1, Trpc5 and Trpc6, and that the levels of their expression are dependent upon the growth stage of the cells. In addition we examined the growth stage dependent expression of the genes encoding Orai1 and Stim1, two proteins that have recently been shown to be involved in SOCE. Our results suggest that the differential expression of Trpc5, Trpc6, Orai1 and Stim1 in quiescent and density-arrested NRK fibroblasts is responsible for the difference in regulation of calcium entry between these cells. Finally, we show that inhibition or potentiation of SOCE and ROCE by pharmacological agents has profound effects on calcium dynamics in NRK fibroblasts. |
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