title: In vitro dose-dependent inhibition of the intracellular spontaneous calcium oscillations in developing hippocampal neurons by ketamine.
abstract: Spatial and temporal abnormalities in the frequency and amplitude of the cytosolic calcium oscillations can impact the normal physiological functions of neuronal cells. Recent studies have shown that ketamine can affect the growth and development and even induce the apoptotic death of neurons. This study used isolated developing hippocampal neurons as its study subjects to observe the effect of ketamine on the intracellular calcium oscillations in developing hippocampal neurons and to further explore its underlying mechanism using Fluo-4-loaded laser scanning confocal microscopy. Using a semi-quantitative method to analyze the spontaneous calcium oscillatory activities, a typical type of calcium oscillation was observed in developing hippocampal neurons. In addition, the administration of NMDA (N-Methyl-D-aspartate) at a concentration of 100 µM increased the calcium oscillation amplitude. The administration of MK801 at a concentration of 40 µM inhibited the amplitude and frequency of the calcium oscillations. Our results demonstrated that an increase in the ketamine concentration, starting from 30 µM, gradually decreased the neuronal calcium oscillation amplitude. The inhibition of the calcium oscillation frequency by 300 µM ketamine was statistically significant, and the neuronal calcium oscillations were completely eliminated with the administration of 3,000 µM Ketamine. The administration of 100, 300, and 1,000 µM NMDA to the 1 mM ketamine-pretreated hippocampal neurons restored the frequency and amplitude of the calcium oscillations in a dose-dependent manner. In fact, a concentration of 1,000 µM NMDA completely reversed the decrease in the calcium oscillation frequency and amplitude that was induced by 1 mM ketamine. This study revealed that ketamine can inhibit the frequency and amplitude of the calcium oscillations in developing hippocampal neurons though the NMDAR (NMDA receptor) in a dose-dependent manner, which might highlight a possible underlying mechanism of ketamine toxicity on the rat hippocampal neurons during development.
authors.affiliation: Department of Anesthesiology, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei Province, China.
authors.forename: Lining
authors.initials: L
authors.lastname: Huang
authors.forename: Yue
authors.initials: Y
authors.lastname: Liu
authors.forename: Pei
authors.initials: P
authors.lastname: Zhang
authors.forename: Rongtian
authors.initials: R
authors.lastname: Kang
authors.forename: Ya
authors.initials: Y
authors.lastname: Liu
authors.forename: Xuze
authors.initials: X
authors.lastname: Li
authors.forename: Lijun
authors.initials: L
authors.lastname: Bo
authors.forename: Zhenming
authors.initials: Z
authors.lastname: Dong
mesh_terms: D000700:Analgesics / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis; D020013:Calcium Signaling / Q000187:drug effects*; D003094:Collagen / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D018507:Gene Expression Regulation, Developmental; D006624:Hippocampus / Q000166:cytology* / Q000196:embryology; D007649:Ketamine / Q000494:pharmacology*; D007797:Laminin / Q000737:chemistry; D018613:Microscopy, Confocal; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D009991:Oscillometry; D011509:Proteoglycans / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D000700:Analgesics; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D016202:N-Methylaspartate; D007649:Ketamine; D003094:Collagen
references.citation: Pain Physician. 2011 Jan-Feb;14(1):83-90
references.pmid: 21267045
references.citation: Anesth Analg. 2005 Jun;100(6):1660-1666
references.pmid: 15920192
references.citation: Nat Neurosci. 2010 Jan;13(1):19-21
references.pmid: 20010820
references.citation: J Neurosci. 2007 Aug 15;27(33):8957-66
references.pmid: 17699677
references.citation: Brain Res. 2004 Apr 23;1006(1):8-17
references.pmid: 15047019
references.citation: EMBO Rep. 2006 Apr;7(4):390-6
references.pmid: 16415789
references.citation: Science. 2000 Feb 11;287(5455):1056-60
references.pmid: 10669420
references.citation: Sci STKE. 2004 Oct 19;2004(255):re16
references.pmid: 15494561
references.citation: Semin Cell Dev Biol. 2001 Feb;12(1):3-10
references.pmid: 11162741
references.citation: Eur J Neurosci. 2011 Apr;33(8):1351-65
references.pmid: 21395862
references.citation: Cell Mol Neurobiol. 2004 Dec;24(6):877-82
references.pmid: 15672688
references.citation: Brain Res. 1992 Nov 20;596(1-2):1-9
references.pmid: 1281742
references.citation: Neuron. 1997 Feb;18(2):243-55
references.pmid: 9052795
references.citation: Neuron. 1996 Aug;17(2):275-85
references.pmid: 8780651
references.citation: Anesthesiology. 1973 Oct;39(4):370-6
references.pmid: 4758343
references.citation: Neuroscience. 1997 Jul;79(2):399-409
references.pmid: 9200724
references.citation: Trends Neurosci. 1997 Nov;20(11):523-9
references.pmid: 9364667
references.citation: Science. 1993 Apr 2;260(5104):95-7
references.pmid: 8096653
references.citation: J Exp Biol. 1999 May;202(Pt 10):1319-26
references.pmid: 10210672
references.citation: Exp Brain Res. 1990;80(2):409-14
references.pmid: 2162781
references.citation: J Physiol. 1997 Mar 1;499 ( Pt 2):291-306
references.pmid: 9080360
references.citation: Int J Dev Neurosci. 2009 Nov;27(7):727-31
references.pmid: 19580862
references.citation: Nature. 1995 Jun 29;375(6534):784-7
references.pmid: 7596410
references.citation: Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1115-22
references.pmid: 11830654
references.citation: Annu Rev Biochem. 2008;77:615-41
references.pmid: 18275379
references.citation: Anesthesiology. 1997 Apr;86(4):903-17
references.pmid: 9105235
references.citation: Neurotoxicol Teratol. 2011 Mar-Apr;33(2):220-30
references.pmid: 21241795
references.citation: Nat Rev Mol Cell Biol. 2000 Oct;1(1):11-21
references.pmid: 11413485
references.citation: J Comp Neurol. 2002 Sep 2;450(4):303-17
references.pmid: 12209845
references.citation: J Neurophysiol. 1991 Aug;66(2):473-84
references.pmid: 1723094
references.citation: Neuron. 1994 Mar;12(3):529-40
references.pmid: 7512349
references.citation: Eur J Neurosci. 1999 Feb;11(2):389-97
references.pmid: 10051739
references.citation: J Neurosci. 1999 Nov 15;19(22):9705-15
references.pmid: 10559380
references.citation: Trends Neurosci. 1994 Mar;17(3):115-8
references.pmid: 7515527
references.citation: Trends Biochem Sci. 2011 Feb;36(2):78-87
references.pmid: 20810284
references.citation: J Neurosci. 2003 Feb 1;23(3):927-36
references.pmid: 12574421
references.citation: Toxicol Sci. 2009 Mar;108(1):149-58
references.pmid: 19126600
references.citation: J Neurosci. 2007 Mar 14;27(11):2846-57
references.pmid: 17360906
references.citation: J Pharmacol Exp Ther. 1992 Nov;263(2):569-78
references.pmid: 1331405
references.citation: Neuron. 1998 Jul;21(1):13-26
references.pmid: 9697848