Luisa Michelle Volk, Jan-Erik Bruun, Sandra Trautmann, Dominique Thomas, Stephanie Schwalm, Josef Pfeilschifter, Dagmar Meyer Zu Heringdorf
{"title":"质膜 Ca2+ ATP 酶在鞘氨醇激酶-1 调节细胞 Ca2+ 平衡中的作用","authors":"Luisa Michelle Volk, Jan-Erik Bruun, Sandra Trautmann, Dominique Thomas, Stephanie Schwalm, Josef Pfeilschifter, Dagmar Meyer Zu Heringdorf","doi":"10.1007/s00424-024-03027-7","DOIUrl":null,"url":null,"abstract":"<p><p>Sphingosine-1-phosphate (S1P) is a ubiquitous lipid mediator, acting via specific G-protein-coupled receptors (GPCR) and intracellularly. Previous work has shown that deletion of S1P lyase caused a chronic elevation of cytosolic [Ca<sup>2+</sup>]<sub>i</sub> and enhanced Ca<sup>2+</sup> storage in mouse embryonic fibroblasts. Here, we studied the role of sphingosine kinase (SphK)-1 in Ca<sup>2+</sup> signaling, using two independently generated EA.hy926 cell lines with stable knockdown of SphK1 (SphK1-KD1/2). Resting [Ca<sup>2+</sup>]<sub>i</sub> and thapsigargin-induced [Ca<sup>2+</sup>]<sub>i</sub> increases were reduced in both SphK1-KD1 and -KD2 cells. Agonist-induced [Ca<sup>2+</sup>]<sub>i</sub> increases, measured in SphK1-KD1, were blunted. In the absence of extracellular Ca<sup>2+</sup>, thapsigargin-induced [Ca<sup>2+</sup>]<sub>i</sub> increases declined rapidly, indicating enhanced removal of Ca<sup>2+</sup> from the cytosol. In agreement, plasma membrane Ca<sup>2+</sup> ATPase (PMCA)-1 and -4 and their auxiliary subunit, basigin, were strongly upregulated. Activation of S1P-GPCR by specific agonists or extracellular S1P did not rescue the effects of SphK1 knockdown, indicating that S1P-GPCR were not involved. Lipid measurements indicated that not only S1P but also dihydro-sphingosine, ceramides, and lactosylceramides were markedly depleted in SphK1-KD2 cells. SphK2 and S1P lyase were upregulated, suggesting enhanced flux via the sphingolipid degradation pathway. Finally, histone acetylation was enhanced in SphK1-KD2 cells, and the histone deacetylase inhibitor, vorinostat, induced upregulation of PMCA1 and basigin on mRNA and protein levels in EA.hy926 cells. These data show for the first time a transcriptional regulation of PMCA1 and basigin by S1P metabolism. It is concluded that SphK1 knockdown in EA.hy926 cells caused long-term alterations in cellular Ca<sup>2+</sup> homeostasis by upregulating PMCA via increased histone acetylation.</p>","PeriodicalId":19954,"journal":{"name":"Pflugers Archiv : European journal of physiology","volume":null,"pages":null},"PeriodicalIF":2.9000,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A role for plasma membrane Ca<sup>2+</sup> ATPases in regulation of cellular Ca<sup>2+</sup> homeostasis by sphingosine kinase-1.\",\"authors\":\"Luisa Michelle Volk, Jan-Erik Bruun, Sandra Trautmann, Dominique Thomas, Stephanie Schwalm, Josef Pfeilschifter, Dagmar Meyer Zu Heringdorf\",\"doi\":\"10.1007/s00424-024-03027-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Sphingosine-1-phosphate (S1P) is a ubiquitous lipid mediator, acting via specific G-protein-coupled receptors (GPCR) and intracellularly. Previous work has shown that deletion of S1P lyase caused a chronic elevation of cytosolic [Ca<sup>2+</sup>]<sub>i</sub> and enhanced Ca<sup>2+</sup> storage in mouse embryonic fibroblasts. Here, we studied the role of sphingosine kinase (SphK)-1 in Ca<sup>2+</sup> signaling, using two independently generated EA.hy926 cell lines with stable knockdown of SphK1 (SphK1-KD1/2). Resting [Ca<sup>2+</sup>]<sub>i</sub> and thapsigargin-induced [Ca<sup>2+</sup>]<sub>i</sub> increases were reduced in both SphK1-KD1 and -KD2 cells. Agonist-induced [Ca<sup>2+</sup>]<sub>i</sub> increases, measured in SphK1-KD1, were blunted. In the absence of extracellular Ca<sup>2+</sup>, thapsigargin-induced [Ca<sup>2+</sup>]<sub>i</sub> increases declined rapidly, indicating enhanced removal of Ca<sup>2+</sup> from the cytosol. In agreement, plasma membrane Ca<sup>2+</sup> ATPase (PMCA)-1 and -4 and their auxiliary subunit, basigin, were strongly upregulated. Activation of S1P-GPCR by specific agonists or extracellular S1P did not rescue the effects of SphK1 knockdown, indicating that S1P-GPCR were not involved. Lipid measurements indicated that not only S1P but also dihydro-sphingosine, ceramides, and lactosylceramides were markedly depleted in SphK1-KD2 cells. SphK2 and S1P lyase were upregulated, suggesting enhanced flux via the sphingolipid degradation pathway. Finally, histone acetylation was enhanced in SphK1-KD2 cells, and the histone deacetylase inhibitor, vorinostat, induced upregulation of PMCA1 and basigin on mRNA and protein levels in EA.hy926 cells. These data show for the first time a transcriptional regulation of PMCA1 and basigin by S1P metabolism. It is concluded that SphK1 knockdown in EA.hy926 cells caused long-term alterations in cellular Ca<sup>2+</sup> homeostasis by upregulating PMCA via increased histone acetylation.</p>\",\"PeriodicalId\":19954,\"journal\":{\"name\":\"Pflugers Archiv : European journal of physiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.9000,\"publicationDate\":\"2024-10-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Pflugers Archiv : European journal of physiology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1007/s00424-024-03027-7\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"PHYSIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pflugers Archiv : European journal of physiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s00424-024-03027-7","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PHYSIOLOGY","Score":null,"Total":0}
A role for plasma membrane Ca2+ ATPases in regulation of cellular Ca2+ homeostasis by sphingosine kinase-1.
Sphingosine-1-phosphate (S1P) is a ubiquitous lipid mediator, acting via specific G-protein-coupled receptors (GPCR) and intracellularly. Previous work has shown that deletion of S1P lyase caused a chronic elevation of cytosolic [Ca2+]i and enhanced Ca2+ storage in mouse embryonic fibroblasts. Here, we studied the role of sphingosine kinase (SphK)-1 in Ca2+ signaling, using two independently generated EA.hy926 cell lines with stable knockdown of SphK1 (SphK1-KD1/2). Resting [Ca2+]i and thapsigargin-induced [Ca2+]i increases were reduced in both SphK1-KD1 and -KD2 cells. Agonist-induced [Ca2+]i increases, measured in SphK1-KD1, were blunted. In the absence of extracellular Ca2+, thapsigargin-induced [Ca2+]i increases declined rapidly, indicating enhanced removal of Ca2+ from the cytosol. In agreement, plasma membrane Ca2+ ATPase (PMCA)-1 and -4 and their auxiliary subunit, basigin, were strongly upregulated. Activation of S1P-GPCR by specific agonists or extracellular S1P did not rescue the effects of SphK1 knockdown, indicating that S1P-GPCR were not involved. Lipid measurements indicated that not only S1P but also dihydro-sphingosine, ceramides, and lactosylceramides were markedly depleted in SphK1-KD2 cells. SphK2 and S1P lyase were upregulated, suggesting enhanced flux via the sphingolipid degradation pathway. Finally, histone acetylation was enhanced in SphK1-KD2 cells, and the histone deacetylase inhibitor, vorinostat, induced upregulation of PMCA1 and basigin on mRNA and protein levels in EA.hy926 cells. These data show for the first time a transcriptional regulation of PMCA1 and basigin by S1P metabolism. It is concluded that SphK1 knockdown in EA.hy926 cells caused long-term alterations in cellular Ca2+ homeostasis by upregulating PMCA via increased histone acetylation.
期刊介绍:
Pflügers Archiv European Journal of Physiology publishes those results of original research that are seen as advancing the physiological sciences, especially those providing mechanistic insights into physiological functions at the molecular and cellular level, and clearly conveying a physiological message. Submissions are encouraged that deal with the evaluation of molecular and cellular mechanisms of disease, ideally resulting in translational research. Purely descriptive papers covering applied physiology or clinical papers will be excluded. Papers on methodological topics will be considered if they contribute to the development of novel tools for further investigation of (patho)physiological mechanisms.