{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T19:19:27Z","timestamp":1761765567303},"reference-count":19,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,7,1]],"date-time":"2012-07-01T00:00:00Z","timestamp":1341100800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CP Neuroscience"],"published-print":{"date-parts":[[2012,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Acetylcholine (ACh) is the neurotransmitter used by cholinergic neurons at the neuromuscular junction, in parasympathetic peripheral nerve terminals, and in important memory\u2010related circuits in the brain, and takes part in other critical functions. ACh is synthesized from choline and acetyl coenzyme A by the enzyme choline acetyltransferase (ChAT). The formation of ACh in cholinergic nerve terminals requires the transport of choline into cells from the extracellular space and the activity of ChAT. High\u2010affinity choline uptake (HACU) represents the majority of choline uptake into the nerve terminal and is the acutely regulated, rate\u2010limiting step in ACh synthesis. HACU can be differentiated from nonspecific choline uptake by inhibition of the choline transporter with hemicholinium. Several methods have been described previously to measure HACU and ChAT activity simultaneously in synaptosomes, but a well\u2010documented protocol for cultured cells is lacking. We describe a procedure for simultaneous measurement of HACU and ChAT in cultured cells by simple radionuclide\u2010based techniques. Using this procedure, we have quantitatively determined HACU and ChAT activity in cholinergically differentiated human neuroblastoma (SK\u2010N\u2010SH) cells. These simple methods can be used for neurochemical and drug discovery studies relevant to several disorders, including Alzheimer's disease, myasthenia gravis, and cardiovascular disease. <jats:italic>Curr. Protoc. Neurosci<\/jats:italic>. 60:7.23.1\u20107.23.16. \u00a9 2012 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/0471142301.ns0723s60","type":"journal-article","created":{"date-parts":[[2012,7,2]],"date-time":"2012-07-02T11:58:12Z","timestamp":1341230292000},"source":"Crossref","is-referenced-by-count":2,"title":["High\u2010Affinity Choline Uptake (HACU) and Choline Acetyltransferase (ChAT) Activity in Neuronal Cultures for Mechanistic and Drug Discovery Studies"],"prefix":"10.1002","volume":"60","author":[{"given":"Balmiki","family":"Ray","sequence":"first","affiliation":[{"name":"Department of Psychiatry, Institute of Psychiatric Research, Indiana University School of Medicine  Indianapolis Indiana"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jason A.","family":"Bailey","sequence":"additional","affiliation":[{"name":"Department of Psychiatry, Institute of Psychiatric Research, Indiana University School of Medicine  Indianapolis Indiana"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jay R.","family":"Simon","sequence":"additional","affiliation":[{"name":"Department of Psychiatry, Institute of Psychiatric Research, Indiana University School of Medicine  Indianapolis Indiana"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Debomoy K.","family":"Lahiri","sequence":"additional","affiliation":[{"name":"Department of Psychiatry, Institute of Psychiatric Research, Indiana University School of Medicine  Indianapolis Indiana"},{"name":"Department of Medical and Molecular Genetics, Indiana University School of Medicine  Indianapolis Indiana"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2012,7]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.2000.3561"},{"key":"e_1_2_9_3_1","first-page":"2643","article-title":"Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture","volume":"33","author":"Biedler J.L.","year":"1973","journal-title":"Cancer Res."},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-2999(88)90654-1"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(93)90011-U"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1150465"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0480349"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0169-328X(99)00150-3"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.90705.2008"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(00)02134-7"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1460-9568.2002.02403.x"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.brainres.2009.07.096"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.3233\/JAD-2010-101374"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.1982.tb12589.x"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1038\/255162a0"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-2952(75)90208-7"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.1976.tb06472.x"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-8993(93)91136-G"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1982.sp014022"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.161288698"}],"container-title":["Current Protocols in Neuroscience"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/0471142301.ns0723s60","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/0471142301.ns0723s60","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/0471142301.ns0723s60","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,20]],"date-time":"2023-08-20T01:07:26Z","timestamp":1692493646000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/0471142301.ns0723s60"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,7]]},"references-count":19,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2012,7]]}},"alternative-id":["10.1002\/0471142301.ns0723s60"],"URL":"https:\/\/doi.org\/10.1002\/0471142301.ns0723s60","archive":["Portico"],"relation":{},"ISSN":["1934-8584","1934-8576"],"issn-type":[{"value":"1934-8584","type":"print"},{"value":"1934-8576","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,7]]}}}