Patch clamp experiments with fixation of the membrane potential were performed on isolated rat prefrontal cortex neurons to study the modulation of ion currents induced by applications of GABA and kainate by GABAB receptors and group I metabotropic glutamate receptors. Blockade of GABAB receptors with the selective antagonist CGP-55845 (5 μM) increased the peak amplitude of the ion current induced by application of GABA (40 μM) by 26 ± 13% (n = 6). The amplitude of the ion current plateau at 14 sec of application of GABA was the same in controls as with blockade of GABAB receptors. The long-term effects of activation of GABAB receptors were studied in terms of the responses to application of GABA and kainate in the presence of baclofen (50 μM), a selective GABAB receptor agonist. Prolonged prior application of baclofen increased the amplitude of responses to application of GABA by 9 ± 2% (n = 8) as compared with controls. Responses to application of kainate did not change in the presence of baclofen. Analogous experiments using trans-ACPD, a selective agonist of groups I and II metabotropic glutamate receptors, did not reveal any changes in responses to application of GABA or kainate. These data point to modulation of GABAA receptors by postsynaptic metabotropic GABAB receptors in rat cerebral cortex neurons
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References
D. V. Amakhin and N. P. Veselkin, “Interaction of the effects of the neurotransmitters glycine and GABA in the central nervous system,” Tsitologiya, 54, No. 6, 469–477 (2012).
D. V. Amakhin, V. A. Popov, A. I. Makliel’, and N. P. Veselkin, “Characteristics of the summation of GABA- and glutamate-mediated ion currents in isolated rat cerebral cortex neurons,” Ros. Fiziol. Zh., 98, No. 12, 1490–1506 (2012).
E. A. Tsvetkin, Yu. A. Polina, A. I. Makliel’, and N. P. Veselkin, “Effects of baclofen on the ionotropic current evoked by application of glycine on spinal cord neurons from the frog Rana temporaria,” Zh. Evolyuts. Biokhim. Fiziol., 44, No. 3, 322–323 (2008).
N. Akaike, M. Kaneda, N. Hori, and O. A. Krishtal, “Blockade of N-methyl-D-aspartate response in enzyme-treated rat hippocampal neurons,” Neurosci. Lett., 87, No. 1–2, 75–79 (1988).
S. Balasubramanian, J. A. Teissere, D. V. Raju, and R. A. Hall, “Heterooligomerization between GABA(A) and GABA(B) receptors regulates GABA(B) receptor trafficking,” J. Biol. Chem., 279, No. 18, 18840–18850 (2004).
A. E. Bandrowski, V. B. Aramakis, S. L. Moore, and J. H. Ashe, “Metabotropic glutamate receptors modify ionotropic glutamate responses in neocortical pyramidal cells and interneurons,” Exp. Brain Res., 136, No. 1, 25–40 (2001).
B. Bettler, K. Kaupmann, J. Mosbacher, and M. Gassmann, “Molecular structure and physiological functions of GABA(B) receptors,” Physiol. Rev., 84, No. 3, 835–867 (2004).
N. G. Bowery, A. L. Hudson, and G. W. Price, “GABA(A) and GABA(B) receptor site distribution in the rat central nervous system,” Neuroscience, 20, No. 2, 365–383 (1987).
M. D. Browning, M. Bureau, E. M. Dudek, and R. W. Olsen, “Protein kinase C and cAMP-dependent protein kinase phosphorylate the beta subunit of the purified gamma-aminobutyric acid A receptor,” Proc. Natl. Acad. Sci. USA, 87, No. 4, 1315–1318 (1990).
J. R. Chalifoux and A. G. Carter, “GABA(B) receptors modulate NMDA receptor calcium signals in dendritic spines,” Neuron, 66, No. 1, 101–113 (2010).
W. M. Connelly, S. J. Fyson, A. C. Errington, et al., “GABA(B) receptors regulate extrasynaptic GABA(A) receptors,” J. Neurosci., 33, No. 9, 3780–3785 (2013).
A. Feigenspan and J. Bormann, “Facilitation of GABAergic signaling in the retina by receptors stimulating adenylate cyclase,” Proc. Natl. Acad. Sci. USA, 91, No. 23, 10893–10897 (1994).
W. Jarolimek, M. Bijak, and U. Misgeld, “Differences in the Cs block of baclofen and 4-aminopyridine induced potassium currents of guinea pig CA3 neurons in vitro,” Synapse, 18, No. 3, 169–177 (1994).
M. Kaneda, H. Nakamura, and N. Akaike, “Mechanical and enzymatic isolation of mammalian CNS neurons,” Neurosci. Res., 5, No. 4, 299–315 (1988).
S. Y. Kawaguchi and T. Hirano, “Signaling cascade regulating longterm potentiation of GABA(A) receptor responsiveness in cerebellar Purkinje neurons,” J. Neurosci., 22, No. 10, 3969–3976 (2002).
A. R. Kay and R. K. Wong, “Isolation of neurons suitable for patch-clamping from adult mammalian central nervous systems,” J. Neurosci. Meth., 16, No. 3, 227–238 (1986).
S. Kellenberger, P. Malherbe, and E. Sigel, “Function of the alpha 1 beta 2 gamma 2S gamma-aminobutyric acid type A receptor is modulated by protein kinase C via multiple phosphorylation sites,” J. Biol. Chem., 267, No. 36, 25660–25553 (1992).
B. J. Krishek, X. Xie, C. Blackstone, et al., “Regulation of GABA(A) receptor function by protein kinase C phosphorylation,” Neuron, 12, No. 5, 1081–1095 (1994).
H. Kubota, S. Katsurabayashi, A. J. Moorhouse, et al., “GABA(B) receptor transduction mechanisms, and cross-talk between protein kinases A and C, in GABAergic terminals synapsing onto neurons of the rat nucleus basalis of Meynert,” J. Physiol., 551, No. 1, 263–276 (2003).
Y. F. Lin, T. P. Angelotti, E. M. Dudek, et al., “Enhancement of recombinant alpha 1 beta 1 gamma 2L gamma-aminobutyric acid A receptor whole-cell currents by protein kinase C is mediated through phosphorylation of both beta 1 and gamma 2L subunits,” Mol. Pharmacol., 50, No. 1, 185–195 (1996).
G. Lopez-Bendito, R. Shigemoto, A. Fairen, and R. Lujan, “Differential distribution of group I metabotropic glutamate receptors during rat cortical development,” Cereb. Cortex, 12, No. 6, 625–638 (2002).
T. K. Machu, J. A. Firestone, and M. D. Browning, “Ca2+/calmodulin-dependent protein kinase II and protein kinase C phosphorylate a synthetic peptide corresponding to a sequence that is specific for the gamma 2L subunit of the GABA(A) receptor,” J. Neurochem., 61, No. 1, 375–377 (1993).
B. J. McDonald, A. Amato, C. N. Connolly, et al., “Adjacent phosphorylation sites on GABA(A) receptor beta subunits determine regulation by cAMP-dependent protein kinase,” Nat. Neurosci., 1, No. 1, 23–28 (1998).
B. J. McDonald and S. J. Moss, “Differential phosphorylation of intracellular domains of gamma-aminobutyric acid type A receptor subunits by calcium/calmodulin type 2-dependent protein kinase and cGMP-dependent protein kinase,” J. Biol. Chem., 269, No. 27, 18111–18117 (1994).
B. J. McDonald and S. J. Moss, “Conserved phosphorylation of the intracellular domains of GABA(A) receptor beta2 and beta3 subunits by cAMP-dependent protein kinase, cGMP-dependent protein kinase protein kinase C and Ca2+/calmodulin type H-dependent protein kinase,” Neuropharmacology, 36, No. 10, 1377–1385 (1997).
S. J. Moss, C. A. Doherty, and R. L. Huganir, “Identification of the cAMP-dependent protein kinase and protein kinase C phosphorylation sites within the major intracellular domains of the beta 1, gamma 2S, and gamma 2L subunits of the gamma-aminobutyric acid type A receptor,” J. Biol. Chem., 267, No. 20, 14470–14476 (1992).
S. J. Moss and T. G. Smart, “Modulation of amino acid-gated ion channels by protein phosphorylation,” Int. Rev. Neurobiol., 39, 1–52 (1996).
S. J. Moss, T. G. Smart, C. D. Blackstone, and R. L. Huganir, “Functional modulation of GABA(A) receptors by cAMP-dependent protein phosphorylation,” Science, 257, No. 5070, 661–665 (1992).
C. M. Niswender and P. J. Conn, “Metabotropic glutamate receptors: physiology, pharmacology, and disease,” Annu. Rev. Pharmacol. Toxicol., 50, 295–322 (2010).
C. L. Padgett and P. A. Slesinger, “GABA(B) receptor coupling to G-proteins and ion channels,” Adv. Pharmacol., 589, 123–147 (2010).
K. D. Parfitt, B. J. Hoffer, and P. C. Bickford-Wimer, “Potentiation of gamma-aminobutyric acid-mediated inhibition by isoproterenol in the cerebellar cortex: receptor specificity,” Neuropharmacology, 29, No. 10, 909–916 (1990).
N. M. Porter, R. E. Twyman, M. D. Uhler, and R. L. Macdonald, “Cyclic AMP-dependent protein kinase decreases GABA(A) receptor current in mouse spinal neurons,” Neuron, 5, No. 6, 789–796 (1990).
M. F. Pozza, N. A. Manuel, M. Steinmann, et al., “Comparison of antagonist potencies at pre- and post-synaptic GABA(B) receptors at inhibitory synapses in the CA 1 region of the rat hippocampus,” Br. J. Pharmacol., 127, No. 1, 211–219 (1999).
A. P. Princivalle, M. N. Pangalos, N. G. Bowery, and R. Spreafico, “Distribution of GABA[B(1a)], GABA[B(1b)] and GABA(B2) receptor protein in cerebral cortex and thalamus of adult rats,” Neuroreport, 12, No. 3, 591–595 (2001).
D. B. Pritchett, H. Sontheimer, B. D. Shivers, et al., “Importance of a novel GABA(A) receptor subunit for benzodiazepine pharmacology,” Nature, 338, No. 6216, 582–585 (1989).
M. Robello, C. Amico, and A. Cupello, “Regulation of GABA(A) receptor in cerebellar granule cells in culture: differential involvement of kinase activities,” Neuroscience, 53, No. 1, 131–138 (1993).
A. Rojas, J. Wetherington, R. Shaw, et al., “Activation of group I metabotropic glutamate receptors potentiates heteromeric kainate receptors,” Mol. Pharmacol., 83, No. 1, 106–121 (2013).
C. Romano, M. A. Sesma, C. T. McDonald, et al., “Distribution of metabotropic glutamate receptor mGluR5 immunoreactivity in rat brain,” J. Comp. Neurol., 355, No. 3, 455–469 (1995).
A. N. Shrivastava, A. Triller, and W. Sieghart, “GABA(A) receptors: post-synaptic co-localization and cross-talk with other receptors,” Front. Cell. Neurosci., 5, 7 (2011).
C. H. Song, X. W. Chen, J. X. Xia, et al., “Modulatory effects of hypocretin-1/orexin-A with glutamate and gamma-aminobutyric acid on freshly isolated pyramidal neurons from the rat prefrontal cortex,” Neurosci. Lett., 399, No. 1–2, 101–105 (2006).
T. Tabata and M. Kano, “GABA(B) receptor-mediated modulation of glutamate signaling in cerebellar Purkinje cells,” Cerebellum, 5, No. 2, 127–133 (2006).
T. Tabata and M. Kano, “GABA(B) receptor-mediated modulation of metabotropic glutamate signaling and synaptic plasticity in central neurons,” Adv. Pharmacol., 58, 149–173 (2010).
W. Tao, M. H. Higgs, W. J. Spain, and C. B. Ransom, “Postsynaptic GABA(B) receptors enhance extrasynaptic GABA(A) receptor function in dentate gyrus granule cells,” J. Neurosci., 33, No. 9, 3738–3743 (2013).
J. P. Tyszkiewicz, Z. Gu, X. Wang, et al., “Group II metabotropic glutamate receptors enhance NMDA receptor currents via a protein kinase C-dependent mechanism in pyramidal neurones of rat prefrontal cortex,” J. Physiol., 554, No. 3, 765–777 (2004).
P. Whiting, R. M. McKernan, and L. L. Iversen, “Another mechanism for creating diversity in gamma-aminobutyrate type A receptors: RNA splicing directs expression of two forms of gamma 2 phosphorylation site,” Proc. Natl. Acad. Sci. USA, 87, No. 24, 9966–9970 (2009).
S. Ymer, A. Draguhn, M. Kohler, et al., “Sequence and expression of a novel GABA(A) receptor alpha subunit,” FEBS Lett., 258, No. 1, 119–122 (1989).
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Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 100, No. 10, pp. 1169–1179, October, 2014.
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Amakhin, D.V., Popov, V.A. & Veselkin, N.P. Modulation of GABA- and Kainate-Mediated Ion Currents in Isolated Rat Cerebral Cortex Neurons by Metabotropic Receptors. Neurosci Behav Physi 46, 430–436 (2016). https://doi.org/10.1007/s11055-016-0254-5
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DOI: https://doi.org/10.1007/s11055-016-0254-5