Contractile Arrest Increases Sarcoplasmic Reticulum Calcium Uptake and SERCA2 Gene Expression in Cultured Neonatal Rat Heart Cells
We developed protocols with intact cultured neonatal rat myocytes to directly evaluate the function of the sarcoplasmic reticulum (SR) Ca-ATPase (or SERCA2), Na-Ca exchange (Na-CaX), and slow Ca transport systems (mitochondria and sarcolemmal Ca-ATPase}. Spontaneously beating control cells were comp...
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| Veröffentlicht in: | Circulation research 1994-05, Vol.74 (5), p.991-997 |
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| creator | Bassani, José W.M Qi, Ming Samarel, Allen M Bers, Donald M |
| description | We developed protocols with intact cultured neonatal rat myocytes to directly evaluate the function of the sarcoplasmic reticulum (SR) Ca-ATPase (or SERCA2), Na-Ca exchange (Na-CaX), and slow Ca transport systems (mitochondria and sarcolemmal Ca-ATPase}. Spontaneously beating control cells were compared with cells cultured for 2 days in the presence of verapamil (verapamil-arrested cells, VA). Intracellular calcium (Cai) transients were measured by use of indo-1 during (1) spontaneous twitches, (2) contractures induced by rapid application of caffeine (CafC, with and without Nao), and (3) twitches induced by brief depolarizations with high [K], solution (K-twitches). We also measured mRNA levels for the SR Ca-ATPase and Na-CaX in the same experimental preparations. The tl/2 for [Ca]i decline when both the SR Ca uptake and Na-CaX were prevented was the same for control and VA cells (≈20 seconds), indicating unaltered slow Ca transport systems. Similarly, there was no significant difference in the t1/2 of CafC when Na-CaX was the main mechanism responsible for [Ca]i decline (t1/2 ≈1.5 seconds), indicating unaltered Na-CaX. Conversely, we found nearly a twofold increase in the rate of [Ca]i decline during K-twitches (control t1/2, 0.84±0.05 seconds; VA t1/2, 0.48±0.06 second; P |
| doi_str_mv | 10.1161/01.RES.74.5.991 |
| format | Article |
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Spontaneously beating control cells were compared with cells cultured for 2 days in the presence of verapamil (verapamil-arrested cells, VA). Intracellular calcium (Cai) transients were measured by use of indo-1 during (1) spontaneous twitches, (2) contractures induced by rapid application of caffeine (CafC, with and without Nao), and (3) twitches induced by brief depolarizations with high [K], solution (K-twitches). We also measured mRNA levels for the SR Ca-ATPase and Na-CaX in the same experimental preparations. The tl/2 for [Ca]i decline when both the SR Ca uptake and Na-CaX were prevented was the same for control and VA cells (≈20 seconds), indicating unaltered slow Ca transport systems. Similarly, there was no significant difference in the t1/2 of CafC when Na-CaX was the main mechanism responsible for [Ca]i decline (t1/2 ≈1.5 seconds), indicating unaltered Na-CaX. Conversely, we found nearly a twofold increase in the rate of [Ca]i decline during K-twitches (control t1/2, 0.84±0.05 seconds; VA t1/2, 0.48±0.06 second; P<.001), indicating an increase in SR Ca-pumping activity in VA cells. This was also reflected by a 56% increase in the peak [Ca]i reached during CafC used to assess maximal SR Ca content (427±49 nmol/L in control versus 665±75 nmol/L in VA cells). In agreement with these functional effects, we found no change in Na-CaX mRNA levels but a marked upregulation of both the SERCA2 mRNA and protein levels in VA cells (to 166±10% and 164±20%, respectively). Thus, verapamil arrest induced an increase in SR Ca uptake (and SERCA2 expression) without affecting the Na-CaX activity (or expression) or the slow Ca transport systems.</description><identifier>ISSN: 0009-7330</identifier><identifier>EISSN: 1524-4571</identifier><identifier>DOI: 10.1161/01.RES.74.5.991</identifier><identifier>PMID: 8156646</identifier><identifier>CODEN: CIRUAL</identifier><language>eng</language><publisher>Hagerstown, MD: American Heart Association, Inc</publisher><subject>Animals ; Animals, Newborn ; Biological and medical sciences ; Caffeine - pharmacology ; Calcium - metabolism ; Calcium-Transporting ATPases - genetics ; Calcium-Transporting ATPases - metabolism ; Carrier Proteins - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation ; Heart ; Major Histocompatibility Complex ; Myocardial Contraction - drug effects ; Myocardium - metabolism ; Rats ; RNA, Messenger - metabolism ; Sarcoplasmic Reticulum - metabolism ; Sodium-Calcium Exchanger ; Verapamil - pharmacology ; Vertebrates: cardiovascular system</subject><ispartof>Circulation research, 1994-05, Vol.74 (5), p.991-997</ispartof><rights>1994 American Heart Association, Inc.</rights><rights>1994 INIST-CNRS</rights><rights>Copyright American Heart Association, Inc. May 1994</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4705-4e540d8df8cf6c50ead993d02da9784f220867c57c1e74e2ed40f3ffa43af3143</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,3674,27905,27906</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4120537$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8156646$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bassani, José W.M</creatorcontrib><creatorcontrib>Qi, Ming</creatorcontrib><creatorcontrib>Samarel, Allen M</creatorcontrib><creatorcontrib>Bers, Donald M</creatorcontrib><title>Contractile Arrest Increases Sarcoplasmic Reticulum Calcium Uptake and SERCA2 Gene Expression in Cultured Neonatal Rat Heart Cells</title><title>Circulation research</title><addtitle>Circ Res</addtitle><description>We developed protocols with intact cultured neonatal rat myocytes to directly evaluate the function of the sarcoplasmic reticulum (SR) Ca-ATPase (or SERCA2), Na-Ca exchange (Na-CaX), and slow Ca transport systems (mitochondria and sarcolemmal Ca-ATPase}. Spontaneously beating control cells were compared with cells cultured for 2 days in the presence of verapamil (verapamil-arrested cells, VA). Intracellular calcium (Cai) transients were measured by use of indo-1 during (1) spontaneous twitches, (2) contractures induced by rapid application of caffeine (CafC, with and without Nao), and (3) twitches induced by brief depolarizations with high [K], solution (K-twitches). We also measured mRNA levels for the SR Ca-ATPase and Na-CaX in the same experimental preparations. The tl/2 for [Ca]i decline when both the SR Ca uptake and Na-CaX were prevented was the same for control and VA cells (≈20 seconds), indicating unaltered slow Ca transport systems. Similarly, there was no significant difference in the t1/2 of CafC when Na-CaX was the main mechanism responsible for [Ca]i decline (t1/2 ≈1.5 seconds), indicating unaltered Na-CaX. Conversely, we found nearly a twofold increase in the rate of [Ca]i decline during K-twitches (control t1/2, 0.84±0.05 seconds; VA t1/2, 0.48±0.06 second; P<.001), indicating an increase in SR Ca-pumping activity in VA cells. This was also reflected by a 56% increase in the peak [Ca]i reached during CafC used to assess maximal SR Ca content (427±49 nmol/L in control versus 665±75 nmol/L in VA cells). In agreement with these functional effects, we found no change in Na-CaX mRNA levels but a marked upregulation of both the SERCA2 mRNA and protein levels in VA cells (to 166±10% and 164±20%, respectively). Thus, verapamil arrest induced an increase in SR Ca uptake (and SERCA2 expression) without affecting the Na-CaX activity (or expression) or the slow Ca transport systems.</description><subject>Animals</subject><subject>Animals, Newborn</subject><subject>Biological and medical sciences</subject><subject>Caffeine - pharmacology</subject><subject>Calcium - metabolism</subject><subject>Calcium-Transporting ATPases - genetics</subject><subject>Calcium-Transporting ATPases - metabolism</subject><subject>Carrier Proteins - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation</subject><subject>Heart</subject><subject>Major Histocompatibility Complex</subject><subject>Myocardial Contraction - drug effects</subject><subject>Myocardium - metabolism</subject><subject>Rats</subject><subject>RNA, Messenger - metabolism</subject><subject>Sarcoplasmic Reticulum - metabolism</subject><subject>Sodium-Calcium Exchanger</subject><subject>Verapamil - pharmacology</subject><subject>Vertebrates: cardiovascular system</subject><issn>0009-7330</issn><issn>1524-4571</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkU1r3DAQhk1pSbdpzz0VRCm9eTP6suzjYrZJILSw25zFVB4TJ7K9lWSSXvvLq7BLDj2IQcyjVzM8RfGRw5rzil8AX--2-7VRa71uGv6qWHEtVKm04a-LFQA0pZES3hbvYrwH4EqK5qw4q7muKlWtir_tPKWALg2e2CYEioldTy4QRopsj8HNB49xHBzbURrc4peRtejdkOvtIeEDMZw6tt_u2o1glzQR2z4dck4c5okNE2sXn5ZAHftO84QJPdthYleEIbGWvI_vizc9-kgfTvW8uP22_dlelTc_Lq_bzU3plAFdKtIKurrra9dXTgNh1zSyA9FhY2rVCwF1ZZw2jpNRJKhT0Mu-RyWxl3nx8-LrMfcQ5t9LXtSOQ3R5ApxoXqI1lZJKVyKDn_8D7-clTHk2K7hQHIQyGbo4Qi7MMQbq7SEMI4Y_loN9VmOB26zGGmW1zWryi0-n2OXXSN0Lf3KR-19OfYwOfR9wckN8wRQXoOXzx-qIPc4-UYgPfnmkYO8Ifbqz2ThI4KLkTaNA51uZj9DyH2g4pes</recordid><startdate>199405</startdate><enddate>199405</enddate><creator>Bassani, José W.M</creator><creator>Qi, Ming</creator><creator>Samarel, Allen M</creator><creator>Bers, Donald M</creator><general>American Heart Association, Inc</general><general>Lippincott</general><general>Lippincott Williams & Wilkins Ovid Technologies</general><scope>IQODW</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>7QP</scope><scope>7T5</scope><scope>7TK</scope><scope>H94</scope><scope>K9.</scope><scope>7X8</scope></search><sort><creationdate>199405</creationdate><title>Contractile Arrest Increases Sarcoplasmic Reticulum Calcium Uptake and SERCA2 Gene Expression in Cultured Neonatal Rat Heart Cells</title><author>Bassani, José W.M ; Qi, Ming ; Samarel, Allen M ; Bers, Donald M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4705-4e540d8df8cf6c50ead993d02da9784f220867c57c1e74e2ed40f3ffa43af3143</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>Animals</topic><topic>Animals, Newborn</topic><topic>Biological and medical sciences</topic><topic>Caffeine - pharmacology</topic><topic>Calcium - metabolism</topic><topic>Calcium-Transporting ATPases - genetics</topic><topic>Calcium-Transporting ATPases - metabolism</topic><topic>Carrier Proteins - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation</topic><topic>Heart</topic><topic>Major Histocompatibility Complex</topic><topic>Myocardial Contraction - drug effects</topic><topic>Myocardium - metabolism</topic><topic>Rats</topic><topic>RNA, Messenger - metabolism</topic><topic>Sarcoplasmic Reticulum - metabolism</topic><topic>Sodium-Calcium Exchanger</topic><topic>Verapamil - pharmacology</topic><topic>Vertebrates: cardiovascular system</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bassani, José W.M</creatorcontrib><creatorcontrib>Qi, Ming</creatorcontrib><creatorcontrib>Samarel, Allen M</creatorcontrib><creatorcontrib>Bers, Donald M</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Circulation research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bassani, José W.M</au><au>Qi, Ming</au><au>Samarel, Allen M</au><au>Bers, Donald M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Contractile Arrest Increases Sarcoplasmic Reticulum Calcium Uptake and SERCA2 Gene Expression in Cultured Neonatal Rat Heart Cells</atitle><jtitle>Circulation research</jtitle><addtitle>Circ Res</addtitle><date>1994-05</date><risdate>1994</risdate><volume>74</volume><issue>5</issue><spage>991</spage><epage>997</epage><pages>991-997</pages><issn>0009-7330</issn><eissn>1524-4571</eissn><coden>CIRUAL</coden><abstract>We developed protocols with intact cultured neonatal rat myocytes to directly evaluate the function of the sarcoplasmic reticulum (SR) Ca-ATPase (or SERCA2), Na-Ca exchange (Na-CaX), and slow Ca transport systems (mitochondria and sarcolemmal Ca-ATPase}. Spontaneously beating control cells were compared with cells cultured for 2 days in the presence of verapamil (verapamil-arrested cells, VA). Intracellular calcium (Cai) transients were measured by use of indo-1 during (1) spontaneous twitches, (2) contractures induced by rapid application of caffeine (CafC, with and without Nao), and (3) twitches induced by brief depolarizations with high [K], solution (K-twitches). We also measured mRNA levels for the SR Ca-ATPase and Na-CaX in the same experimental preparations. The tl/2 for [Ca]i decline when both the SR Ca uptake and Na-CaX were prevented was the same for control and VA cells (≈20 seconds), indicating unaltered slow Ca transport systems. Similarly, there was no significant difference in the t1/2 of CafC when Na-CaX was the main mechanism responsible for [Ca]i decline (t1/2 ≈1.5 seconds), indicating unaltered Na-CaX. Conversely, we found nearly a twofold increase in the rate of [Ca]i decline during K-twitches (control t1/2, 0.84±0.05 seconds; VA t1/2, 0.48±0.06 second; P<.001), indicating an increase in SR Ca-pumping activity in VA cells. This was also reflected by a 56% increase in the peak [Ca]i reached during CafC used to assess maximal SR Ca content (427±49 nmol/L in control versus 665±75 nmol/L in VA cells). In agreement with these functional effects, we found no change in Na-CaX mRNA levels but a marked upregulation of both the SERCA2 mRNA and protein levels in VA cells (to 166±10% and 164±20%, respectively). Thus, verapamil arrest induced an increase in SR Ca uptake (and SERCA2 expression) without affecting the Na-CaX activity (or expression) or the slow Ca transport systems.</abstract><cop>Hagerstown, MD</cop><pub>American Heart Association, Inc</pub><pmid>8156646</pmid><doi>10.1161/01.RES.74.5.991</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Circulation research, 1994-05, Vol.74 (5), p.991-997 |
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| source | MEDLINE; American Heart Association Journals; Journals@Ovid Complete; EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Animals, Newborn Biological and medical sciences Caffeine - pharmacology Calcium - metabolism Calcium-Transporting ATPases - genetics Calcium-Transporting ATPases - metabolism Carrier Proteins - metabolism Fundamental and applied biological sciences. Psychology Gene Expression Regulation Heart Major Histocompatibility Complex Myocardial Contraction - drug effects Myocardium - metabolism Rats RNA, Messenger - metabolism Sarcoplasmic Reticulum - metabolism Sodium-Calcium Exchanger Verapamil - pharmacology Vertebrates: cardiovascular system |
| title | Contractile Arrest Increases Sarcoplasmic Reticulum Calcium Uptake and SERCA2 Gene Expression in Cultured Neonatal Rat Heart Cells |
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