Synergistic effects of agonists and two-pore-domain potassium channels on secretory responses of human pancreatic duct cells Capan-1
Mechanisms of synergistic agonist stimulation and modulation of the electrochemical driving force for anion secretion are still not fully explored in human pancreatic duct epithelial cells. The first objective of this study was therefore to test whether combined agonist stimulation augments anion tr...
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| Veröffentlicht in: | Pflügers Archiv 2023-03, Vol.475 (3), p.361-379 |
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| creator | Sørensen, Christiane E. Trauzold, Anna Christensen, Nynne M. Tawfik, Doaa Szczepanowski, Monika Novak, Ivana |
| description | Mechanisms of synergistic agonist stimulation and modulation of the electrochemical driving force for anion secretion are still not fully explored in human pancreatic duct epithelial cells. The first objective of this study was therefore to test whether combined agonist stimulation augments anion transport responses in the Capan-1 monolayer model of human pancreatic duct epithelium. The second objective was to test the influence of H
+
,K
+
-ATPase inhibition on anion transport in Capan-1 monolayers. The third objective was to analyze the expression and function of K
+
channels in Capan-1, which could support anion secretion and cooperate with H
+
,K
+
-ATPases in pH and potassium homeostasis. The human pancreatic adenocarcinoma cell line Capan-1 was cultured conventionally or as polarized monolayers that were analyzed by Ussing chamber electrophysiological recordings. Single-cell intracellular calcium was assayed with Fura-2. mRNA isolated from Capan-1 was analyzed by use of the nCounter assay or RT-PCR. Protein expression was assessed by immunofluorescence and western blot analyses. Combined stimulation with different physiological agonists enhanced anion transport responses compared to single agonist stimulation. The responsiveness of Capan-1 cells to histamine was also revealed in these experiments. The H
+
,K
+
-ATPase inhibitor omeprazole reduced carbachol- and riluzole-induced anion transport responses. Transcript analyses revealed abundant TASK-2, TWIK-1, TWIK-2, TASK-5, K
Ca3.1
, and KCNQ1 mRNA expression. KCNE1 mRNA and TREK-1, TREK-2, TASK-2, and KCNQ1 protein expression were also shown. This study shows that the Capan-1 model recapitulates key physiological aspects of a bicarbonate-secreting epithelium and constitutes a valuable model for functional studies on human pancreatic duct epithelium. |
| doi_str_mv | 10.1007/s00424-022-02782-9 |
| format | Article |
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+
,K
+
-ATPase inhibition on anion transport in Capan-1 monolayers. The third objective was to analyze the expression and function of K
+
channels in Capan-1, which could support anion secretion and cooperate with H
+
,K
+
-ATPases in pH and potassium homeostasis. The human pancreatic adenocarcinoma cell line Capan-1 was cultured conventionally or as polarized monolayers that were analyzed by Ussing chamber electrophysiological recordings. Single-cell intracellular calcium was assayed with Fura-2. mRNA isolated from Capan-1 was analyzed by use of the nCounter assay or RT-PCR. Protein expression was assessed by immunofluorescence and western blot analyses. Combined stimulation with different physiological agonists enhanced anion transport responses compared to single agonist stimulation. The responsiveness of Capan-1 cells to histamine was also revealed in these experiments. The H
+
,K
+
-ATPase inhibitor omeprazole reduced carbachol- and riluzole-induced anion transport responses. Transcript analyses revealed abundant TASK-2, TWIK-1, TWIK-2, TASK-5, K
Ca3.1
, and KCNQ1 mRNA expression. KCNE1 mRNA and TREK-1, TREK-2, TASK-2, and KCNQ1 protein expression were also shown. This study shows that the Capan-1 model recapitulates key physiological aspects of a bicarbonate-secreting epithelium and constitutes a valuable model for functional studies on human pancreatic duct epithelium.</description><identifier>ISSN: 0031-6768</identifier><identifier>EISSN: 1432-2013</identifier><identifier>DOI: 10.1007/s00424-022-02782-9</identifier><identifier>PMID: 36534232</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Adenocarcinoma ; Adenocarcinoma - metabolism ; Adenosine Triphosphatases - metabolism ; Agonists ; Bicarbonates ; Bicarbonates - metabolism ; Biomedical and Life Sciences ; Biomedicine ; Calcium (intracellular) ; Carbachol ; Cell Biology ; Epithelial cells ; Epithelial Cells - metabolism ; Epithelium ; Fura-2 ; Gene expression ; Homeostasis ; Human Physiology ; Humans ; Hydrogen ; Immunofluorescence ; Ion Channels ; Ion Channels, Receptors and Transporters ; Ions ; KCNQ1 protein ; Molecular Medicine ; Neurosciences ; Omeprazole ; Pancreas ; Pancreatic Ducts ; Pancreatic Neoplasms - metabolism ; Physiology ; Potassium ; Potassium channels (voltage-gated) ; Protein expression ; Receptors ; Receptors and Transporters ; RNA, Messenger - metabolism</subject><ispartof>Pflügers Archiv, 2023-03, Vol.475 (3), p.361-379</ispartof><rights>The Author(s) 2022</rights><rights>2022. The Author(s).</rights><rights>The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-df7bc410592440e1d0b6f26cd7e139711eca4e310168b2822cb4d1f2a43d59353</citedby><cites>FETCH-LOGICAL-c474t-df7bc410592440e1d0b6f26cd7e139711eca4e310168b2822cb4d1f2a43d59353</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00424-022-02782-9$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00424-022-02782-9$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36534232$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Sørensen, Christiane E.</creatorcontrib><creatorcontrib>Trauzold, Anna</creatorcontrib><creatorcontrib>Christensen, Nynne M.</creatorcontrib><creatorcontrib>Tawfik, Doaa</creatorcontrib><creatorcontrib>Szczepanowski, Monika</creatorcontrib><creatorcontrib>Novak, Ivana</creatorcontrib><title>Synergistic effects of agonists and two-pore-domain potassium channels on secretory responses of human pancreatic duct cells Capan-1</title><title>Pflügers Archiv</title><addtitle>Pflugers Arch - Eur J Physiol</addtitle><addtitle>Pflugers Arch</addtitle><description>Mechanisms of synergistic agonist stimulation and modulation of the electrochemical driving force for anion secretion are still not fully explored in human pancreatic duct epithelial cells. The first objective of this study was therefore to test whether combined agonist stimulation augments anion transport responses in the Capan-1 monolayer model of human pancreatic duct epithelium. The second objective was to test the influence of H
+
,K
+
-ATPase inhibition on anion transport in Capan-1 monolayers. The third objective was to analyze the expression and function of K
+
channels in Capan-1, which could support anion secretion and cooperate with H
+
,K
+
-ATPases in pH and potassium homeostasis. The human pancreatic adenocarcinoma cell line Capan-1 was cultured conventionally or as polarized monolayers that were analyzed by Ussing chamber electrophysiological recordings. Single-cell intracellular calcium was assayed with Fura-2. mRNA isolated from Capan-1 was analyzed by use of the nCounter assay or RT-PCR. Protein expression was assessed by immunofluorescence and western blot analyses. Combined stimulation with different physiological agonists enhanced anion transport responses compared to single agonist stimulation. The responsiveness of Capan-1 cells to histamine was also revealed in these experiments. The H
+
,K
+
-ATPase inhibitor omeprazole reduced carbachol- and riluzole-induced anion transport responses. Transcript analyses revealed abundant TASK-2, TWIK-1, TWIK-2, TASK-5, K
Ca3.1
, and KCNQ1 mRNA expression. KCNE1 mRNA and TREK-1, TREK-2, TASK-2, and KCNQ1 protein expression were also shown. This study shows that the Capan-1 model recapitulates key physiological aspects of a bicarbonate-secreting epithelium and constitutes a valuable model for functional studies on human pancreatic duct epithelium.</description><subject>Adenocarcinoma</subject><subject>Adenocarcinoma - metabolism</subject><subject>Adenosine Triphosphatases - metabolism</subject><subject>Agonists</subject><subject>Bicarbonates</subject><subject>Bicarbonates - metabolism</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Calcium (intracellular)</subject><subject>Carbachol</subject><subject>Cell Biology</subject><subject>Epithelial cells</subject><subject>Epithelial Cells - metabolism</subject><subject>Epithelium</subject><subject>Fura-2</subject><subject>Gene expression</subject><subject>Homeostasis</subject><subject>Human Physiology</subject><subject>Humans</subject><subject>Hydrogen</subject><subject>Immunofluorescence</subject><subject>Ion Channels</subject><subject>Ion Channels, Receptors and Transporters</subject><subject>Ions</subject><subject>KCNQ1 protein</subject><subject>Molecular Medicine</subject><subject>Neurosciences</subject><subject>Omeprazole</subject><subject>Pancreas</subject><subject>Pancreatic Ducts</subject><subject>Pancreatic Neoplasms - metabolism</subject><subject>Physiology</subject><subject>Potassium</subject><subject>Potassium channels (voltage-gated)</subject><subject>Protein expression</subject><subject>Receptors</subject><subject>Receptors and Transporters</subject><subject>RNA, Messenger - metabolism</subject><issn>0031-6768</issn><issn>1432-2013</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9Uc9rFDEUDmKxa_Uf8CABz6nJS3YycxFkUSsUPLSeQyZ5sztlJxmTjGXv_uGm3Vr10sPjwft-PfgIeSP4ueBcv8-cK1CMA9TRLbDuGVkJJYEBF_I5WXEuBWt0056SlznfcM5BtfCCnMpmLRVIWJFfV4eAaTvmMjqKw4CuZBoHarcx1GOmNnhabiObY0Lm42THQOdYbM7jMlG3syHgvkoCzegSlpgONGGeY8h477RbJlslNlTU3qX4xRXqcF9VG1vvTLwiJ4PdZ3z9sM_I98-frjcX7PLbl6-bj5fMKa0K84PunRJ83YFSHIXnfTNA47xGITstBDqrUAoumraHFsD1yosBrJJ-3cm1PCMfjr7z0k_oHYaS7N7MaZxsOphoR_M_Esad2cafput42zSiGrx7MEjxx4K5mJu4pFB_NqC1kqoDqSsLjiyXYs4Jh8cEwc1dc-bYnKnNmfvmTFdFb__97VHyp6pKkEdCrlDYYvqb_YTtbx8lpwY</recordid><startdate>20230301</startdate><enddate>20230301</enddate><creator>Sørensen, Christiane E.</creator><creator>Trauzold, Anna</creator><creator>Christensen, Nynne M.</creator><creator>Tawfik, Doaa</creator><creator>Szczepanowski, Monika</creator><creator>Novak, Ivana</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><scope>C6C</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>3V.</scope><scope>7QP</scope><scope>7TK</scope><scope>7TS</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>5PM</scope></search><sort><creationdate>20230301</creationdate><title>Synergistic effects of agonists and two-pore-domain potassium channels on secretory responses of human pancreatic duct cells Capan-1</title><author>Sørensen, Christiane E. ; Trauzold, Anna ; Christensen, Nynne M. ; Tawfik, Doaa ; Szczepanowski, Monika ; Novak, Ivana</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-df7bc410592440e1d0b6f26cd7e139711eca4e310168b2822cb4d1f2a43d59353</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Adenocarcinoma</topic><topic>Adenocarcinoma - metabolism</topic><topic>Adenosine Triphosphatases - metabolism</topic><topic>Agonists</topic><topic>Bicarbonates</topic><topic>Bicarbonates - metabolism</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Calcium (intracellular)</topic><topic>Carbachol</topic><topic>Cell Biology</topic><topic>Epithelial cells</topic><topic>Epithelial Cells - metabolism</topic><topic>Epithelium</topic><topic>Fura-2</topic><topic>Gene expression</topic><topic>Homeostasis</topic><topic>Human Physiology</topic><topic>Humans</topic><topic>Hydrogen</topic><topic>Immunofluorescence</topic><topic>Ion Channels</topic><topic>Ion Channels, Receptors and Transporters</topic><topic>Ions</topic><topic>KCNQ1 protein</topic><topic>Molecular Medicine</topic><topic>Neurosciences</topic><topic>Omeprazole</topic><topic>Pancreas</topic><topic>Pancreatic Ducts</topic><topic>Pancreatic Neoplasms - metabolism</topic><topic>Physiology</topic><topic>Potassium</topic><topic>Potassium channels (voltage-gated)</topic><topic>Protein expression</topic><topic>Receptors</topic><topic>Receptors and Transporters</topic><topic>RNA, Messenger - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sørensen, Christiane E.</creatorcontrib><creatorcontrib>Trauzold, Anna</creatorcontrib><creatorcontrib>Christensen, Nynne M.</creatorcontrib><creatorcontrib>Tawfik, Doaa</creatorcontrib><creatorcontrib>Szczepanowski, Monika</creatorcontrib><creatorcontrib>Novak, Ivana</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Physical Education Index</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection (ProQuest)</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Pflügers Archiv</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sørensen, Christiane E.</au><au>Trauzold, Anna</au><au>Christensen, Nynne M.</au><au>Tawfik, Doaa</au><au>Szczepanowski, Monika</au><au>Novak, Ivana</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synergistic effects of agonists and two-pore-domain potassium channels on secretory responses of human pancreatic duct cells Capan-1</atitle><jtitle>Pflügers Archiv</jtitle><stitle>Pflugers Arch - Eur J Physiol</stitle><addtitle>Pflugers Arch</addtitle><date>2023-03-01</date><risdate>2023</risdate><volume>475</volume><issue>3</issue><spage>361</spage><epage>379</epage><pages>361-379</pages><issn>0031-6768</issn><eissn>1432-2013</eissn><abstract>Mechanisms of synergistic agonist stimulation and modulation of the electrochemical driving force for anion secretion are still not fully explored in human pancreatic duct epithelial cells. The first objective of this study was therefore to test whether combined agonist stimulation augments anion transport responses in the Capan-1 monolayer model of human pancreatic duct epithelium. The second objective was to test the influence of H
+
,K
+
-ATPase inhibition on anion transport in Capan-1 monolayers. The third objective was to analyze the expression and function of K
+
channels in Capan-1, which could support anion secretion and cooperate with H
+
,K
+
-ATPases in pH and potassium homeostasis. The human pancreatic adenocarcinoma cell line Capan-1 was cultured conventionally or as polarized monolayers that were analyzed by Ussing chamber electrophysiological recordings. Single-cell intracellular calcium was assayed with Fura-2. mRNA isolated from Capan-1 was analyzed by use of the nCounter assay or RT-PCR. Protein expression was assessed by immunofluorescence and western blot analyses. Combined stimulation with different physiological agonists enhanced anion transport responses compared to single agonist stimulation. The responsiveness of Capan-1 cells to histamine was also revealed in these experiments. The H
+
,K
+
-ATPase inhibitor omeprazole reduced carbachol- and riluzole-induced anion transport responses. Transcript analyses revealed abundant TASK-2, TWIK-1, TWIK-2, TASK-5, K
Ca3.1
, and KCNQ1 mRNA expression. KCNE1 mRNA and TREK-1, TREK-2, TASK-2, and KCNQ1 protein expression were also shown. This study shows that the Capan-1 model recapitulates key physiological aspects of a bicarbonate-secreting epithelium and constitutes a valuable model for functional studies on human pancreatic duct epithelium.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>36534232</pmid><doi>10.1007/s00424-022-02782-9</doi><tpages>19</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0031-6768 |
| ispartof | Pflügers Archiv, 2023-03, Vol.475 (3), p.361-379 |
| issn | 0031-6768 1432-2013 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_9908661 |
| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | Adenocarcinoma Adenocarcinoma - metabolism Adenosine Triphosphatases - metabolism Agonists Bicarbonates Bicarbonates - metabolism Biomedical and Life Sciences Biomedicine Calcium (intracellular) Carbachol Cell Biology Epithelial cells Epithelial Cells - metabolism Epithelium Fura-2 Gene expression Homeostasis Human Physiology Humans Hydrogen Immunofluorescence Ion Channels Ion Channels, Receptors and Transporters Ions KCNQ1 protein Molecular Medicine Neurosciences Omeprazole Pancreas Pancreatic Ducts Pancreatic Neoplasms - metabolism Physiology Potassium Potassium channels (voltage-gated) Protein expression Receptors Receptors and Transporters RNA, Messenger - metabolism |
| title | Synergistic effects of agonists and two-pore-domain potassium channels on secretory responses of human pancreatic duct cells Capan-1 |
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