{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T09:20:08Z","timestamp":1778664008441,"version":"3.51.4"},"reference-count":90,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2015,10,27]],"date-time":"2015-10-27T00:00:00Z","timestamp":1445904000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100005339","name":"American Academy of Neurology","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100005339","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100005331","name":"American Brain Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100005331","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["5R01NS40109\u201014"],"award-info":[{"award-number":["5R01NS40109\u201014"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"publisher","award":["R01NS74772\u201004"],"award-info":[{"award-number":["R01NS74772\u201004"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Ann Clin Transl Neurol"],"published-print":{"date-parts":[[2015,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Objective<\/jats:title><jats:p>Although benzodiazepines and other <jats:styled-content style=\"fixed-case\">GABA<jats:sub>A<\/jats:sub><\/jats:styled-content> receptors allosteric modulators are used to treat neonatal seizures, their efficacy may derive from actions on subcortical structures. Side effects of benzodiazepines in nonseizing human neonates include myoclonus, seizures, and abnormal movements. Excitatory actions of <jats:styled-content style=\"fixed-case\">GABA<\/jats:styled-content> may underlie both side effects and reduced anticonvulsant activity of benzodiazepines. Neocortical organotypic slice cultures were used to study: (1) spontaneous cortical epileptiform activity during early development; (2) developmental changes in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> and (3) whether diazepam's anticonvulsant effect correlated with neuronal [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Epileptiform activity in neocortical organotypic slice cultures was measured by field potential recordings. Cl<jats:sup>\u2212<\/jats:sup> changes during development were assessed by multiphoton imaging of neurons transgenically expressing a Cl\u2010sensitive fluorophore. Clinically relevant concentrations of diazepam were used to test the anticonvulsant effectiveness at ages corresponding to premature neonates through early infancy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>(1) Neocortical organotypic slices at days in vitro 5 (<jats:styled-content style=\"fixed-case\">DIV<\/jats:styled-content>5) exhibited spontaneous epileptiform activity. (2) Epileptiform event duration decreased with age. (3) There was a progressive decrease in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> over the same age range. (4) Diazepam was ineffective in decreasing epileptiform activity at <jats:styled-content style=\"fixed-case\">DIV<\/jats:styled-content>5\u20106, but progressively more effective at older ages through <jats:styled-content style=\"fixed-case\">DIV<\/jats:styled-content>15. (5) At <jats:styled-content style=\"fixed-case\">DIV<\/jats:styled-content>5\u20106, diazepam worsened epileptiform activity in 50% of the slices.<\/jats:p><\/jats:sec><jats:sec><jats:title>Interpretation<\/jats:title><jats:p>The neocortical organotypic slice is a useful model to study spontaneous epileptiform activity. Decreasing [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> during development correlates with decreasing duration of spontaneous epileptiform activity and increasing anticonvulsant efficacy of diazepam. We provide a potential explanation for the reports of seizures and myoclonus induction by benzodiazepines in newborn human neonates and the limited electrographic efficacy of benzodiazepines for the treatment of neonatal seizures.<\/jats:p><\/jats:sec>","DOI":"10.1002\/acn3.259","type":"journal-article","created":{"date-parts":[[2015,10,27]],"date-time":"2015-10-27T12:36:23Z","timestamp":1445949383000},"page":"1055-1070","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["Diazepam effect during early neonatal development correlates with neuronal Cl<sup>\u2212<\/sup>"],"prefix":"10.1002","volume":"2","author":[{"given":"Joseph","family":"Glykys","sequence":"first","affiliation":[{"name":"Department of Neurology Massachusetts General Hospital Boston Massachusetts"},{"name":"Harvard Medical School Boston Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin J.","family":"Staley","sequence":"additional","affiliation":[{"name":"Department of Neurology Massachusetts General Hospital Boston Massachusetts"},{"name":"Harvard Medical School Boston Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,10,27]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1987.sp016493"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.16-20-06414.1996"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(91)90243-S"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4695(19990615)39:4<558::AID-NEU9>3.0.CO;2-5"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00017.2006"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2009.08.022"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm1301"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2013.00020"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5169-06.2007"},{"issue":"3","key":"e_1_2_8_11_1","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1113\/jphysiol.2004.062471","article-title":"Cl\u2212 uptake promoting depolarizing GABA actions in immature rat neocortical neurones is mediated by NKCC1","volume":"557","author":"Yamada J","year":"2004","journal-title":"J Physiol"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.10983"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrn920"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncpneuro0883"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4695(19971120)33:6<781::AID-NEU6>3.0.CO;2-5"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.181342398"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.15.8556"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcn.2009.07.012"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.conb.2010.08.013"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1245423"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2014.275529"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00962.2005"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1002149"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2013.04.005"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1097\/00019052-200304000-00010"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1136\/adc.64.4_Spec_No.459"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199908123410704"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2013.01.001"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1684\/epd.2014.0690"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1212\/01.WNL.0000106944.59990.E6"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1177\/0883073807299858"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1097\/00002826-199619020-00005"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1345\/aph.1A328"},{"key":"e_1_2_8_34_1","first-page":"98","article-title":"Clonic movements following lorazepam administration in full\u2010term infants","volume":"152","author":"Chess PR","year":"1998","journal-title":"Arch Pediatr Adolesc Med"},{"key":"e_1_2_8_35_1","first-page":"146","article-title":"Stereotypic movements after lorazepam administration in premature neonates: a series and review of the literature","volume":"15","author":"Sexson WR","year":"1995","journal-title":"J Perinatol"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1542\/peds.104.3.579"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1159\/000244123"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbd.2011.11.001"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1097\/WNP.0b013e3181fe0709"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1111\/cns.12369"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1016\/0165-0270(89)90051-4"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.1987.57.3.869"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.1999.81.1.399"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.1988.59.1.259"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1088\/1741-2560\/4\/2\/001"},{"key":"e_1_2_8_46_1","doi-asserted-by":"crossref","first-page":"1018","DOI":"10.1152\/jn.1993.70.3.1018","article-title":"On the synchronous activity induced by 4\u2010aminopyridine in the CA3 subfield of juvenile rat hippocampus","volume":"70","author":"Avoli M","year":"1993","journal-title":"J Neurophysiol"},{"key":"e_1_2_8_47_1","doi-asserted-by":"crossref","first-page":"7873","DOI":"10.1523\/JNEUROSCI.23-21-07873.2003","article-title":"Transition from interictal to ictal activity in limbic networks in vitro","volume":"23","author":"Dzhala VI","year":"2003","journal-title":"J Neurosci"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn1125"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(03)12658-X"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)00056-8"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-2236(97)01122-3"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.90402.2008"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.5139-11.2012"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbd.2012.05.016"},{"key":"e_1_2_8_55_1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awp217"},{"key":"e_1_2_8_56_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2005.09.014"},{"key":"e_1_2_8_57_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1528-1167.2009.02339.x"},{"key":"e_1_2_8_58_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.78.11.7180"},{"key":"e_1_2_8_59_1","doi-asserted-by":"publisher","DOI":"10.1038\/271563a0"},{"key":"e_1_2_8_60_1","doi-asserted-by":"publisher","DOI":"10.1038\/269342a0"},{"key":"e_1_2_8_61_1","first-page":"187","volume-title":"Antiepileptic drugs","author":"Anderson G","year":"2002"},{"key":"e_1_2_8_62_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF02244106"},{"key":"e_1_2_8_63_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.4007-14.2015"},{"key":"e_1_2_8_64_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-444-52898-8.00004-5"},{"key":"e_1_2_8_65_1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/73.3.392"},{"key":"e_1_2_8_66_1","doi-asserted-by":"publisher","DOI":"10.1016\/0013-4694(64)90149-X"},{"key":"e_1_2_8_67_1","doi-asserted-by":"publisher","DOI":"10.1590\/S0004-282X2012000200014"},{"key":"e_1_2_8_68_1","doi-asserted-by":"publisher","DOI":"10.1016\/0896-6273(95)90008-X"},{"key":"e_1_2_8_69_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1528-1167.2007.01295.x"},{"key":"e_1_2_8_70_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.23-05-01840.2003"},{"key":"e_1_2_8_71_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-8993(78)90447-X"},{"key":"e_1_2_8_72_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1996.sp021505"},{"key":"e_1_2_8_73_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-394316-3.00006-5"},{"key":"e_1_2_8_74_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.23-10-04134.2003"},{"key":"e_1_2_8_75_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00970.2011"},{"key":"e_1_2_8_76_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.00616.2004"},{"key":"e_1_2_8_77_1","doi-asserted-by":"crossref","first-page":"8069","DOI":"10.1523\/JNEUROSCI.20-21-08069.2000","article-title":"Potassium\u2010coupled chloride cotransport controls intracellular chloride in rat neocortical pyramidal neurons","volume":"20","author":"DeFazio R","year":"2000","journal-title":"J Neurosci"},{"key":"e_1_2_8_78_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.3378-07.2008"},{"key":"e_1_2_8_79_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.1997.273.5.C1516"},{"key":"e_1_2_8_80_1","doi-asserted-by":"publisher","DOI":"10.1038\/pr.2012.7"},{"key":"e_1_2_8_81_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.21229"},{"key":"e_1_2_8_82_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.siny.2013.05.008"},{"key":"e_1_2_8_83_1","first-page":"1","article-title":"Bumetanide for the treatment of seizures in newborn babies with hypoxic ischaemic encephalopathy (NEMO): an open\u2010label, dose finding, and feasibility phase 1\/2 trial","volume":"4422","author":"Pressler RM","year":"2015","journal-title":"Lancet Neurol"},{"key":"e_1_2_8_84_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrneurol.2015.74"},{"key":"e_1_2_8_85_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1995.sp020649"},{"key":"e_1_2_8_86_1","doi-asserted-by":"publisher","DOI":"10.1038\/378075a0"},{"key":"e_1_2_8_87_1","doi-asserted-by":"publisher","DOI":"10.1093\/brain\/awh181"},{"key":"e_1_2_8_88_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tins.2007.05.003"},{"key":"e_1_2_8_89_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.4105-14.2015"},{"key":"e_1_2_8_90_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.12-11-04151.1992"},{"key":"e_1_2_8_91_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2007.134460"}],"container-title":["Annals of Clinical and Translational Neurology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Facn3.259","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/acn3.259","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/acn3.259","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/acn3.259","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,5]],"date-time":"2023-10-05T03:35:55Z","timestamp":1696476955000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/acn3.259"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,10,27]]},"references-count":90,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2015,12]]}},"alternative-id":["10.1002\/acn3.259"],"URL":"https:\/\/doi.org\/10.1002\/acn3.259","archive":["Portico"],"relation":{},"ISSN":["2328-9503","2328-9503"],"issn-type":[{"value":"2328-9503","type":"print"},{"value":"2328-9503","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,10,27]]},"assertion":[{"value":"2015-09-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-09-16","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-10-27","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}