title: Changes of calcium channel mRNA, protein and current in NG108-15 cells after cell differentiation.
abstract: Based on the characteristics of differentiated NG108-15 cells (cell membrane excitability, acetylcholine release, and activities of choline acetyltransferase and acetylcholinesterase), NG108-15 cells are extensively used to explore neuronal functions as a cholinergic cell line. In the present study, differentiation-induced alterations of voltage-gated Ca(2+) channel mRNA, protein, and current were investigated in the NG108-15 cells. Real-time PCR, Western blot, and whole-cell patch-clamp data showed that differentiation caused mRNA, protein, and ion current changes of all Ca(2+) channel subunits. However, the changes of mRNA, protein, and ion current are inconsistent in all Ca(2+) channel subunits. Especially, P/Q- and R-type Ca(2+) channel proteins do not form the functional P/Q- and R-type Ca(2+) channels even if the mRNA and protein of P/Q- and R-type Ca(2+) channels can be detected in NG108-15 cells. These results indicate that differentiation can modulate gene transcription, protein translation, and post-translation of the Ca(2+) channels to induce the alteration of the Ca(2+) ion currents in NG108-15 cells. From these data, we understand that combining real-time PCR, Western blot, and patch-clamp techniques can comprehensively unveil the modulation of the Ca(2+) channels.
authors.affiliation: Department of Emergency Medicine, University of Nebraska Medical Center, Omaha, NE 68198-5850, USA.
authors.forename: Jinxu
authors.initials: J
authors.lastname: Liu
authors.forename: Huiyin
authors.initials: H
authors.lastname: Tu
authors.forename: Dongze
authors.initials: D
authors.lastname: Zhang
authors.forename: Yu-Long
authors.initials: YL
authors.lastname: Li
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D059329:Cholinergic Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005786:Gene Expression Regulation; D017136:Ion Transport; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D014158:Transcription, Genetic
chemical_list: D015220:Calcium Channels; D012333:RNA, Messenger