{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,29]],"date-time":"2025-09-29T11:58:39Z","timestamp":1759147119841},"reference-count":40,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2004,6,2]],"date-time":"2004-06-02T00:00:00Z","timestamp":1086134400000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Eur J of Neuroscience"],"published-print":{"date-parts":[[2004,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The homeostasis of intracellular Cl<jats:sup>\u2212<\/jats:sup> concentration ([Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub>) is critical for neuronal function, including \u03b3\u2010aminobutyric acid (GABA)ergic synaptic transmission. Here, we investigated activity\u2010dependent changes in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> using a transgenetically expressed Cl<jats:sup>\u2212<\/jats:sup>\u2010sensitive enhanced yellow\u2010fluorescent protein (EYFP) in cultures of mouse hippocampal neurons. Application of glutamate (100\u2003\u00b5<jats:sc>m<\/jats:sc> for 3\u2003min) in a bath perfusion to cell cultures of various days <jats:italic>in vitro<\/jats:italic> (DIV) revealed a decrease in EYFP fluorescence. The EYFP signal increased in amplitude with increasing DIV, reaching a maximal response after 7\u2003DIV. Glutamate application resulted in a slight neuronal acidification. Although EYFP fluorescence is sensitive to pH, EYFP signals were virtually abolished in Cl<jats:sup>\u2212<\/jats:sup>\u2010free solution, demonstrating that the EYFP signal represented an increase in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub>. Similar to glutamate, a rise in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> was also induced by specific ionotropic glutamate receptor agonists and by increasing extracellular [K<jats:sup>+<\/jats:sup>], indicating that an increase in driving force for Cl<jats:sup>\u2212<\/jats:sup> suffices to increase [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub>. To elucidate the membrane mechanisms mediating the Cl<jats:sup>\u2212<\/jats:sup> influx, a series of blockers of ion channels and transporters were tested. The glutamate\u2010induced increase in [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> was resistant to furosemide, bumetanide and 4,4\u2032\u2010diisothiocyanato\u2010stilbene\u20102,2\u2032\u2010disulphonic acid (DIDS), was reduced by bicuculline to about 80% of control responses, and was antagonized by niflumic acid (NFA) and 5\u2010nitro\u20102\u2010(3\u2010phenylpropylamino)benzoic acid (NPPB). We conclude that membrane depolarization increases [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> via several pathways involving NFA\u2010 and NPPB\u2010sensitive anion channels and GABA<jats:sub>A<\/jats:sub> receptors, but not through furosemide\u2010, bumetanide\u2010 or DIDS\u2010sensitive Cl<jats:sup>\u2212<\/jats:sup> transporters. The present study highlights the vulnerability of [Cl<jats:sup>\u2212<\/jats:sup>]<jats:sub>i<\/jats:sub> homeostasis after membrane depolarization in neurons.<\/jats:p>","DOI":"10.1111\/j.0953-816x.2004.03422.x","type":"journal-article","created":{"date-parts":[[2004,6,2]],"date-time":"2004-06-02T15:28:38Z","timestamp":1086190118000},"page":"2915-2922","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Glutamate\u2010induced elevations in intracellular chloride concentration in hippocampal cell cultures derived from EYFP\u2010expressing mice"],"prefix":"10.1111","volume":"19","author":[{"given":"Jennifer E.","family":"Slemmer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shinichi","family":"Matsushita","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chris I.","family":"De Zeeuw","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John T.","family":"Weber","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas","family":"Kn\u00f6pfel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,6,2]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.75.4.403"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/281315a0"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1978.sp012387"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.23-12-05061.2003"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1177\/1073858402238514"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi9902182"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(01)02561-3"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00341-5"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1080\/01616412.1998.11740495"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.13-11-04690.1993"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1471-4159.1998.70062500.x"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1471-4159.1998.71041396.x"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.275.9.6047"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1152\/jn.2000.84.1.281"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(98)77870-1"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)00056-8"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.0953-816x.2001.01837.x"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(90)92062-I"},{"key":"e_1_2_6_20_1","first-page":"7P","article-title":"New design for a chloride\u2010sensitive micro\u2010electrode","volume":"231","author":"Neild T.O.","year":"1973","journal-title":"J. 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