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Mechanisms of heterogeneity of calcium response to kainate and neuronal types in rat cortical primary culture

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Mechanisms of heterogeneity of calcium response to kainate and neuronal types in rat cortical primary culture

Content Provider	Springer Nature Link
Author	Abushik, P. A. Bolshakov, A. E. Sibarov, D. A. Antov, S. M.
Copyright Year	2011
Abstract	The dynamics of intracellular Ca$^{2+}$ signal in response to NMDA (N-methyl-D-aspartate, 30 μM) or KA (kainite, 30 μM), its dependence on extracellular Ca$^{2+}$ and the mechanisms of KA-triggered Ca$^{2+}$ entry into neurons have been tested in neurons of rat cortical primary cultures. The level of intracellular free Ca$^{2+}$ concentrations ([Ca$^{2+}$]$_{i}$) was evaluated on Leica SP5 MF confocal microscope using Fluo-3 fluorescent dye, which resolves changes in [Ca$^{2+}$]$_{i}$ in the micromolar range. The dynamics of [Ca$^{2+}$]$_{i}$ increase in response to NMDA and KA was different but in both cases the [Ca$^{2+}$]$_{i}$ increase required the presence of Ca$^{2+}$ in the extracellular solution. The neuronal population was found to be heterogeneous, based on the response to KA applied together with either L-type calcium channel blocker nifedipine (3 μM) or IEM-1460 (3 μM), a blocker of Ca$^{2+}$-permeable AMPAR (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor) lacking GluR2 subunit. Experiments exhibited three types of calcium responses, characteristically belonging to interneurons (expressing Ca$^{2+}$-permeable AMPAR), pyramidal neurons (with AMPAR containing GluR2, making them impermeable to Ca$^{2+}$), and intermediate type of cells expressing both AMPAR types. Thus, we have demonstrated the role of AMPAR and L-type calcium channels in KA-triggered Ca$^{2+}$ entry into neurons. The dynamics of [Ca$^{2+}$]$_{i}$ during the KA treatment was shown to depend on subunit composition of particular AMPAR subtype expressed in neurons. The data suggest that neuronal types existing in adult cortical tissue are probably presented in primary culture, too.
Starting Page	92
Ending Page	100
Page Count	9
File Format	PDF
ISSN	19907478
Journal	Biochemistry (Moscow) Supplement Series A: Membrane and Cell Biology
Volume Number	5
Issue Number	1
e-ISSN	19907494
Language	English
Publisher	SP MAIK Nauka/Interperiodica
Publisher Date	2011-03-05
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	kainite intracellular calcium neurons ion channels glutamate receptors Cell Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Biophysics

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