Acute reversible SERCA blockade facilitates or blocks exocytosis, respectively in mouse or bovine chromaffin cells
Pre-blockade of the sarco-endoplasmic reticulum (ER) calcium ATPase (SERCA) with irreversible thapsigargin depresses exocytosis in adrenal bovine chromaffin cells (BCCs). Distinct expression of voltage-dependent Ca 2+ -channel subtypes and of the Ca 2+ -induced Ca 2+ release (CICR) mechanism in BCCs...
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| Veröffentlicht in: | Pflügers Archiv 2021-02, Vol.473 (2), p.273-286 |
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| creator | Martínez-Ramírez, Carmen Gil-Gómez, Irene G. de Diego, Antonio M. García, Antonio G. |
| description | Pre-blockade of the sarco-endoplasmic reticulum (ER) calcium ATPase (SERCA) with irreversible thapsigargin depresses exocytosis in adrenal bovine chromaffin cells (BCCs). Distinct expression of voltage-dependent Ca
2+
-channel subtypes and of the Ca
2+
-induced Ca
2+
release (CICR) mechanism in BCCs versus mouse chromaffin cells (MCCs) has been described. We present a parallel study on the effects of the acute SERCA blockade with reversible cyclopizonic acid (CPA), to repeated pulsing with acetylcholine (ACh) at short (15 s) and long intervals (60 s) at 37 °C, allowing the monitoring of the initial size of a ready-release vesicle pool (RRP) and its depletion and recovery in subsequent stimuli. We found (i) strong depression of exocytosis upon ACh pulsing at 15-s intervals and slower depression at 60-s intervals in both cell types; (ii) facilitation of exocytosis upon acute SERCA inhibition, with back to depression upon CPA washout in MCCs; (iii) blockade of exocytosis upon acute SERCA inhibition and pronounced rebound of exocytosis upon CPA washout in BCCs; (iv) basal [Ca
2+
]
c
elevation upon stimulation with ACh at short intervals (but not at long intervals) in both cell types; and (v) augmentation of basal [Ca
2+
]
c
and inhibition of peak [Ca
2+
]
c
amplitude upon CPA treatment in both cell types, with milder effects upon stimulation at 60-s intervals. These results are compatible with the view that while in MCCs the uptake of Ca
2+
via SERCA contributes to the mitigation of physiological ACh triggered secretion, in BCCs the uptake of Ca
2+
into the ER facilitates such responses likely potentiating a Ca
2+
-induced Ca
2+
release mechanism. These drastic differences in the regulation of ACh-triggered secretion at 37 °C may help to understand different patterns of the regulation of exocytosis by the circulation of Ca
2+
at a functional ER Ca
2+
store. |
| doi_str_mv | 10.1007/s00424-020-02483-1 |
| format | Article |
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2+
-channel subtypes and of the Ca
2+
-induced Ca
2+
release (CICR) mechanism in BCCs versus mouse chromaffin cells (MCCs) has been described. We present a parallel study on the effects of the acute SERCA blockade with reversible cyclopizonic acid (CPA), to repeated pulsing with acetylcholine (ACh) at short (15 s) and long intervals (60 s) at 37 °C, allowing the monitoring of the initial size of a ready-release vesicle pool (RRP) and its depletion and recovery in subsequent stimuli. We found (i) strong depression of exocytosis upon ACh pulsing at 15-s intervals and slower depression at 60-s intervals in both cell types; (ii) facilitation of exocytosis upon acute SERCA inhibition, with back to depression upon CPA washout in MCCs; (iii) blockade of exocytosis upon acute SERCA inhibition and pronounced rebound of exocytosis upon CPA washout in BCCs; (iv) basal [Ca
2+
]
c
elevation upon stimulation with ACh at short intervals (but not at long intervals) in both cell types; and (v) augmentation of basal [Ca
2+
]
c
and inhibition of peak [Ca
2+
]
c
amplitude upon CPA treatment in both cell types, with milder effects upon stimulation at 60-s intervals. These results are compatible with the view that while in MCCs the uptake of Ca
2+
via SERCA contributes to the mitigation of physiological ACh triggered secretion, in BCCs the uptake of Ca
2+
into the ER facilitates such responses likely potentiating a Ca
2+
-induced Ca
2+
release mechanism. These drastic differences in the regulation of ACh-triggered secretion at 37 °C may help to understand different patterns of the regulation of exocytosis by the circulation of Ca
2+
at a functional ER Ca
2+
store.</description><identifier>ISSN: 0031-6768</identifier><identifier>EISSN: 1432-2013</identifier><identifier>DOI: 10.1007/s00424-020-02483-1</identifier><identifier>PMID: 33108514</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Acetylcholine ; Acetylcholine - pharmacology ; Animals ; Biomedical and Life Sciences ; Biomedicine ; Ca2+-transporting ATPase ; Calcium (reticular) ; Calcium - metabolism ; Calcium channels (voltage-gated) ; Calcium influx ; Calcium Signaling - drug effects ; Catecholamines - metabolism ; Cattle ; Cell Biology ; Cells, Cultured ; Chromaffin cells ; Chromaffin Cells - drug effects ; Chromaffin Cells - enzymology ; Endoplasmic reticulum ; Endoplasmic Reticulum - drug effects ; Endoplasmic Reticulum - enzymology ; Enzyme Inhibitors - pharmacology ; Exocytosis ; Exocytosis - drug effects ; Human Physiology ; Indoles - pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; Molecular Medicine ; Neurosciences ; Receptors ; Sarcoplasmic Reticulum Calcium-Transporting ATPases - antagonists & inhibitors ; Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism ; Secretion ; Signaling and Cell Physiology ; Species Specificity ; Thapsigargin ; Thapsigargin - pharmacology ; Time Factors</subject><ispartof>Pflügers Archiv, 2021-02, Vol.473 (2), p.273-286</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020</rights><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2020.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c375t-f1afe6b14d6a83afa9ee6364357b6862f600021211616c2c5c1f7e7ab027c88b3</citedby><cites>FETCH-LOGICAL-c375t-f1afe6b14d6a83afa9ee6364357b6862f600021211616c2c5c1f7e7ab027c88b3</cites><orcidid>0000-0002-3557-3330</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00424-020-02483-1$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00424-020-02483-1$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/33108514$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Martínez-Ramírez, Carmen</creatorcontrib><creatorcontrib>Gil-Gómez, Irene</creatorcontrib><creatorcontrib>G. de Diego, Antonio M.</creatorcontrib><creatorcontrib>García, Antonio G.</creatorcontrib><title>Acute reversible SERCA blockade facilitates or blocks exocytosis, respectively in mouse or bovine chromaffin cells</title><title>Pflügers Archiv</title><addtitle>Pflugers Arch - Eur J Physiol</addtitle><addtitle>Pflugers Arch</addtitle><description>Pre-blockade of the sarco-endoplasmic reticulum (ER) calcium ATPase (SERCA) with irreversible thapsigargin depresses exocytosis in adrenal bovine chromaffin cells (BCCs). Distinct expression of voltage-dependent Ca
2+
-channel subtypes and of the Ca
2+
-induced Ca
2+
release (CICR) mechanism in BCCs versus mouse chromaffin cells (MCCs) has been described. We present a parallel study on the effects of the acute SERCA blockade with reversible cyclopizonic acid (CPA), to repeated pulsing with acetylcholine (ACh) at short (15 s) and long intervals (60 s) at 37 °C, allowing the monitoring of the initial size of a ready-release vesicle pool (RRP) and its depletion and recovery in subsequent stimuli. We found (i) strong depression of exocytosis upon ACh pulsing at 15-s intervals and slower depression at 60-s intervals in both cell types; (ii) facilitation of exocytosis upon acute SERCA inhibition, with back to depression upon CPA washout in MCCs; (iii) blockade of exocytosis upon acute SERCA inhibition and pronounced rebound of exocytosis upon CPA washout in BCCs; (iv) basal [Ca
2+
]
c
elevation upon stimulation with ACh at short intervals (but not at long intervals) in both cell types; and (v) augmentation of basal [Ca
2+
]
c
and inhibition of peak [Ca
2+
]
c
amplitude upon CPA treatment in both cell types, with milder effects upon stimulation at 60-s intervals. These results are compatible with the view that while in MCCs the uptake of Ca
2+
via SERCA contributes to the mitigation of physiological ACh triggered secretion, in BCCs the uptake of Ca
2+
into the ER facilitates such responses likely potentiating a Ca
2+
-induced Ca
2+
release mechanism. These drastic differences in the regulation of ACh-triggered secretion at 37 °C may help to understand different patterns of the regulation of exocytosis by the circulation of Ca
2+
at a functional ER Ca
2+
store.</description><subject>Acetylcholine</subject><subject>Acetylcholine - pharmacology</subject><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Ca2+-transporting ATPase</subject><subject>Calcium (reticular)</subject><subject>Calcium - metabolism</subject><subject>Calcium channels (voltage-gated)</subject><subject>Calcium influx</subject><subject>Calcium Signaling - drug effects</subject><subject>Catecholamines - metabolism</subject><subject>Cattle</subject><subject>Cell Biology</subject><subject>Cells, Cultured</subject><subject>Chromaffin cells</subject><subject>Chromaffin Cells - drug effects</subject><subject>Chromaffin Cells - enzymology</subject><subject>Endoplasmic reticulum</subject><subject>Endoplasmic Reticulum - drug effects</subject><subject>Endoplasmic Reticulum - enzymology</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Exocytosis</subject><subject>Exocytosis - drug effects</subject><subject>Human Physiology</subject><subject>Indoles - pharmacology</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Inbred C57BL</subject><subject>Molecular Medicine</subject><subject>Neurosciences</subject><subject>Receptors</subject><subject>Sarcoplasmic Reticulum Calcium-Transporting ATPases - antagonists & inhibitors</subject><subject>Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism</subject><subject>Secretion</subject><subject>Signaling and Cell Physiology</subject><subject>Species Specificity</subject><subject>Thapsigargin</subject><subject>Thapsigargin - pharmacology</subject><subject>Time Factors</subject><issn>0031-6768</issn><issn>1432-2013</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><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>eNp9kVFLHDEUhUOp1NX2D_RBAn3pg1Nzk0wyfVwWq4JQUPscMtmbNnZmsiYzi_vvzTraQh_6EAK53zn3hEPIR2BfgDF9lhmTXFaMs3JkIyp4QxYgBa84A_GWLBgTUCmtmkNylPM9Y3uMvyOHQgBrapALkpZuGpEm3GLKoe2Q3p7frJa07aL7bddIvXWhC6MdMdOY5vdM8TG63RhzyKdFmzfoxrDFbkfDQPs4ZXxm4zYMSN2vFHvrfRk57Lr8nhx422X88HIfkx_fzu9Wl9X194ur1fK6ckLXY-XBelQtyLWyjbDefkVUQklR61Y1inu1_w9wAAXKcVc78Bq1bRnXrmlacUw-z76bFB8mzKPpQ94nsAOWiIbLugYthRIF_fQPeh-nNJR0hWpYLZXSdaH4TLkUc07ozSaF3qadAWb2jZi5EVMaMc-NGCiikxfrqe1x_UfyWkEBxAzkMhp-Yvq7-z-2T2T5lm8</recordid><startdate>20210201</startdate><enddate>20210201</enddate><creator>Martínez-Ramírez, Carmen</creator><creator>Gil-Gómez, Irene</creator><creator>G. de Diego, Antonio M.</creator><creator>García, Antonio G.</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><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>7X8</scope><orcidid>https://orcid.org/0000-0002-3557-3330</orcidid></search><sort><creationdate>20210201</creationdate><title>Acute reversible SERCA blockade facilitates or blocks exocytosis, respectively in mouse or bovine chromaffin cells</title><author>Martínez-Ramírez, Carmen ; Gil-Gómez, Irene ; G. de Diego, Antonio M. ; García, Antonio G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c375t-f1afe6b14d6a83afa9ee6364357b6862f600021211616c2c5c1f7e7ab027c88b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Acetylcholine</topic><topic>Acetylcholine - pharmacology</topic><topic>Animals</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Ca2+-transporting ATPase</topic><topic>Calcium (reticular)</topic><topic>Calcium - metabolism</topic><topic>Calcium channels (voltage-gated)</topic><topic>Calcium influx</topic><topic>Calcium Signaling - drug effects</topic><topic>Catecholamines - metabolism</topic><topic>Cattle</topic><topic>Cell Biology</topic><topic>Cells, Cultured</topic><topic>Chromaffin cells</topic><topic>Chromaffin Cells - drug effects</topic><topic>Chromaffin Cells - enzymology</topic><topic>Endoplasmic reticulum</topic><topic>Endoplasmic Reticulum - drug effects</topic><topic>Endoplasmic Reticulum - enzymology</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Exocytosis</topic><topic>Exocytosis - drug effects</topic><topic>Human Physiology</topic><topic>Indoles - pharmacology</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Molecular Medicine</topic><topic>Neurosciences</topic><topic>Receptors</topic><topic>Sarcoplasmic Reticulum Calcium-Transporting ATPases - antagonists & inhibitors</topic><topic>Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism</topic><topic>Secretion</topic><topic>Signaling and Cell Physiology</topic><topic>Species Specificity</topic><topic>Thapsigargin</topic><topic>Thapsigargin - pharmacology</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Martínez-Ramírez, Carmen</creatorcontrib><creatorcontrib>Gil-Gómez, Irene</creatorcontrib><creatorcontrib>G. de Diego, Antonio M.</creatorcontrib><creatorcontrib>García, Antonio G.</creatorcontrib><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</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>MEDLINE - Academic</collection><jtitle>Pflügers Archiv</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Martínez-Ramírez, Carmen</au><au>Gil-Gómez, Irene</au><au>G. de Diego, Antonio M.</au><au>García, Antonio G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Acute reversible SERCA blockade facilitates or blocks exocytosis, respectively in mouse or bovine chromaffin cells</atitle><jtitle>Pflügers Archiv</jtitle><stitle>Pflugers Arch - Eur J Physiol</stitle><addtitle>Pflugers Arch</addtitle><date>2021-02-01</date><risdate>2021</risdate><volume>473</volume><issue>2</issue><spage>273</spage><epage>286</epage><pages>273-286</pages><issn>0031-6768</issn><eissn>1432-2013</eissn><abstract>Pre-blockade of the sarco-endoplasmic reticulum (ER) calcium ATPase (SERCA) with irreversible thapsigargin depresses exocytosis in adrenal bovine chromaffin cells (BCCs). Distinct expression of voltage-dependent Ca
2+
-channel subtypes and of the Ca
2+
-induced Ca
2+
release (CICR) mechanism in BCCs versus mouse chromaffin cells (MCCs) has been described. We present a parallel study on the effects of the acute SERCA blockade with reversible cyclopizonic acid (CPA), to repeated pulsing with acetylcholine (ACh) at short (15 s) and long intervals (60 s) at 37 °C, allowing the monitoring of the initial size of a ready-release vesicle pool (RRP) and its depletion and recovery in subsequent stimuli. We found (i) strong depression of exocytosis upon ACh pulsing at 15-s intervals and slower depression at 60-s intervals in both cell types; (ii) facilitation of exocytosis upon acute SERCA inhibition, with back to depression upon CPA washout in MCCs; (iii) blockade of exocytosis upon acute SERCA inhibition and pronounced rebound of exocytosis upon CPA washout in BCCs; (iv) basal [Ca
2+
]
c
elevation upon stimulation with ACh at short intervals (but not at long intervals) in both cell types; and (v) augmentation of basal [Ca
2+
]
c
and inhibition of peak [Ca
2+
]
c
amplitude upon CPA treatment in both cell types, with milder effects upon stimulation at 60-s intervals. These results are compatible with the view that while in MCCs the uptake of Ca
2+
via SERCA contributes to the mitigation of physiological ACh triggered secretion, in BCCs the uptake of Ca
2+
into the ER facilitates such responses likely potentiating a Ca
2+
-induced Ca
2+
release mechanism. These drastic differences in the regulation of ACh-triggered secretion at 37 °C may help to understand different patterns of the regulation of exocytosis by the circulation of Ca
2+
at a functional ER Ca
2+
store.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>33108514</pmid><doi>10.1007/s00424-020-02483-1</doi><tpages>14</tpages><orcidid>https://orcid.org/0000-0002-3557-3330</orcidid></addata></record> |
| fulltext | fulltext |
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| ispartof | Pflügers Archiv, 2021-02, Vol.473 (2), p.273-286 |
| issn | 0031-6768 1432-2013 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_2455174363 |
| source | MEDLINE; SpringerLink Journals - AutoHoldings |
| subjects | Acetylcholine Acetylcholine - pharmacology Animals Biomedical and Life Sciences Biomedicine Ca2+-transporting ATPase Calcium (reticular) Calcium - metabolism Calcium channels (voltage-gated) Calcium influx Calcium Signaling - drug effects Catecholamines - metabolism Cattle Cell Biology Cells, Cultured Chromaffin cells Chromaffin Cells - drug effects Chromaffin Cells - enzymology Endoplasmic reticulum Endoplasmic Reticulum - drug effects Endoplasmic Reticulum - enzymology Enzyme Inhibitors - pharmacology Exocytosis Exocytosis - drug effects Human Physiology Indoles - pharmacology Male Mice Mice, Inbred C57BL Molecular Medicine Neurosciences Receptors Sarcoplasmic Reticulum Calcium-Transporting ATPases - antagonists & inhibitors Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism Secretion Signaling and Cell Physiology Species Specificity Thapsigargin Thapsigargin - pharmacology Time Factors |
| title | Acute reversible SERCA blockade facilitates or blocks exocytosis, respectively in mouse or bovine chromaffin cells |
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