title: Cultured embryonic hippocampal neurons deficient in glucocorticoid (GC) receptor: a novel model for studying nongenomic effects of GC in the neural system.
abstract: Glucocorticoid (GC) acts through both genomic and nongenomic mechanisms. It affects the structure and function of the central nervous system, especially the hippocampus. Here we report an in vitro culture system that can yield embryonic hippocampal neurons deficient in the expression of GC receptor as demonstrated by immunoblotting, immunocytochemistry, and RT-PCR. Owing to this unique feature, those neuron preparations can serve as an ideal model for studying the nongenomic actions of GC on neural cells. In this study, we found that the Erk1/2, c-Jun N-terminal kinase (JNK), and p38 MAPKs were activated in these neurons by BSA-conjugated corticosterone within 15 min of treatment. This activation was not blocked by RU38486, spironolactone, or cycloheximide. Therefore, it is concluded that the activation of MAPKs observed here was due to the nongenomic action of GC. Furthermore, a 24-h incubation with corticosterone at concentrations ranged from 10(-11)-10(-5) M did not have an effect on the viability of GC receptor-deficient neurons.
authors.affiliation: Institute of Neuroscience, Department of Neurobiology, Second Military Medical University, 800 XiangYin Road, Shanghai 200433, People's Republic of China.
authors.forename: Lin
authors.initials: L
authors.lastname: Xiao
authors.forename: Aiqun
authors.initials: A
authors.lastname: Qi
authors.forename: Yizhang
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D003345:Corticosterone / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology* / Q000196:embryology; D006727:Hormone Antagonists / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000502:physiology*; D015735:Mifepristone / Q000494:pharmacology; D000451:Mineralocorticoid Receptor Antagonists / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D010766:Phosphorylation / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000235:genetics* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013148:Spironolactone / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D016895:Culture Media, Serum-Free; D006727:Hormone Antagonists; D000451:Mineralocorticoid Receptor Antagonists; D011500:Protein Synthesis Inhibitors; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; C439264:bovine serum albumin-corticosterone conjugate; D012710:Serum Albumin, Bovine; D013148:Spironolactone; D015735:Mifepristone; D003513:Cycloheximide; D048031:JNK Mitogen-Activated Protein Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D003345:Corticosterone