Suppression of NMDA Receptor Function Using Antisense DNA Blocks Ocular Dominance Plasticity While Preserving Visual Responses

1998 ◽  
Vol 80 (3) ◽  
pp. 1021-1032 ◽  
Author(s):  
Elizabeth B. Roberts ◽  
M. Alex Meredith ◽  
Ary S. Ramoa

Roberts, Elizabeth B., M. Alex Meredith, and Ary S. Ramoa. Suppression of NMDA receptor function using antisense DNA blocks ocular dominance plasticity while preserving visual responses. J. Neurophysiol. 80: 1021–1032, 1998. Pioneering work has shown that pharmacological blockade of the N-methyl-d-aspartate (NMDA) receptor channel reduces ocular dominance plasticity. However, the results also show that doses of NMDA receptor antagonists that have an effect on ocular dominance plasticity profoundly reduce sensory responses and disrupt stimulus selectivity of cortical cells. It is, therefore, not possible to determine whether effects of NMDA receptor blockade on visual plasticity result from a specific role of NMDA receptors or from the reduction in sensory response. We have used an alternate approach to examine this question. We performed knockdown experiments using antisense oligodeoxynucleotides (ODNs) complementary to mRNA coding the NR1 subunit of the NMDA receptor. After 5 days of antisense, but not sense, ODN treatment NMDA receptor–mediated synaptic transmission was reduced markedly relative to the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor response, as indicated by whole cell patch-clamp recordings in the cortical slice preparation. This suppression of NMDA receptor–mediated currents was due to a selective reduction in the NR1 protein near the injection site relative to the untreated hemisphere in the same animal, as indicated by immunocytochemistry and Western blotting. In contrast, AMPA receptors were not affected by the antisense ODN treatment indicating specificity of effects. Another major effect of this treatment was to decrease ocular dominance plasticity. Ferrets that were monocularly deprived 1 wk during the antisense ODN treatment had ocular dominance histograms similar to those found in untreated, nondeprived animals. In contrast, ferrets treated with sense ODN and monocularly deprived had ocular dominance histograms resembling those of untreated, monocularly deprived animals. The effects on ocular dominance plasticity did not result from a disruption of sensory responses because maximum responses as well as orientation and direction selectivity of cortical cells were not affected by the treatment. In conclusion, the present results show that antisense techniques can accomplish more selective manipulations of cortical function than is possible with traditional pharmacological agents. Use of this approach also provides unambiguous evidence for a specific role of NMDA receptors in visual plasticity.

2015 ◽  
Vol 67 (3) ◽  
pp. 490-493 ◽  
Author(s):  
Małgorzata Wolak ◽  
Agata Siwek ◽  
Bernadeta Szewczyk ◽  
Ewa Poleszak ◽  
Beata Bystrowska ◽  
...  

2017 ◽  
Vol 28 (4) ◽  
pp. 1183-1194 ◽  
Author(s):  
M Hadi Saiepour ◽  
Rogier Min ◽  
Willem Kamphuis ◽  
J Alexander Heimel ◽  
Christiaan N Levelt

2021 ◽  
Author(s):  
Kai Langer ◽  
Adriana Mueller-Laengle ◽  
Jannik Wempe ◽  
Bodo Laube

Ionotropic glutamate receptors (iGluRs) are tetrameric ligand-gated ion channels that play a crucial role in excitatory synaptic transmission in the central nervous system. Each subunit contributes with three transmembrane domains (M1, M3, and M4) and a pore loop (M2) forming the channel pore. Recent studies suggest that the architecture of all eukaryotic iGluRs derives from a common prokaryotic ancestral receptor that lacks M4 and consists only of the transmembrane domain segments M1-M3. Although a crucial contribution of M4 to the assembly and trafficking of iGluRs is suspected, the role of this additionally evolved domain in receptor function remains controversial. Here, we investigated how deletions and mutations of M4 in members of the NMDA receptor subfamily, the conventional heteromeric GluN1/GluN2 and glycine-gated GluN1/GluN3 NMDA receptors, affect expression and function in Xenopus oocytes. We show that deletion of M4 in the GluN1, GluN2, or GluN3 subunit, despite retained receptor assembly and cell surface expression, results in nonfunctional membrane receptors. Coexpression of the corresponding M4 domains as an isolated peptide in M4-deleted receptors rescued receptor function of GluN1/GluN2A NMDARs without altering the affinity of glutamate or glycine. Substitution of non-conserved residues and insertion of interhelical disulfide bridges confirmed the proximity of positions M813 and F817 in M4 of GluN1 to residues of the TMs of neighboring subunits. Electrophysiological analyses of agonist-induced receptor function and its modulation by the neurosteroid pregnenolone sulfate (PS) at mutations of the GluN1-M4/GluN2/3-TM interface indicate a crucial role of interdomain interactions in the functional coupling of M4 to the nuclear receptor and the modulatory effect of PS. Our results show that although the M4 domains in NMDA receptors are not important for receptor assembly and surface expression, residues at the subunit interface are substantially involved in M4 recognition to the core receptor and regulation of PS efficacy. Because mutations in the M4 of GluN1 specifically resulted in loss of PS-induced inhibition of NMDA receptor currents, our results point to distinct roles of M4s in NMDA receptor modulation and highlight the importance of evolutionarily newly evolved M4s for selective in vivo modulation of glutamate- and glycine-activated NMDA receptors by steroids.


Author(s):  
Joseph P. Steiner ◽  
Kathryn B. Payne ◽  
Christopher Drummond Main ◽  
Sabrina D'Alfonso ◽  
Kirsten X. Jacobsen ◽  
...  

Background:Previously we showed that 6-hydroxydopamine lesions of the substantia nigra eliminate corticostriatal LTP and that the neuroimmunolophilin ligand (NIL), GPI-1046, restores LTP.Methods:We used cDNA microarrays to determine what mRNAs may be over- or under-expressed in response to lesioning and/or GPI-1046 treatment. Patch clamp recordings were performed to investigate changes in NMDA channel function before and after treatments.Results:We found that 51 gene products were differentially expressed. Among these we found that GPI-1046 treatment up-regulated presenilin-1 (PS-1) mRNA abundance. This finding was confirmed using QPCR. PS-1 protein was also shown to be over-expressed in the striatum of lesioned/GPI-1046-treated rats. As PS-1 has been implicated in controlling NMDA-receptor function and LTP is reduced by lesioning we assayed NMDA mediated synaptic activity in striatal brain slices. The lesion-induced reduction of dopaminergic innervation was accompanied by the near complete loss of NDMA receptor-mediated synaptic transmission between the cortex and striatum. GPI-1046 treatment of the lesioned rats restored NMDA-mediated synaptic transmission but not the dopaminergic innervation. Restoration of NDMA channel function was apparently specific as the sodium channel current density was also reduced due to lesioning but GPI-1046 did not reverse this effect. We also found that restoration of NMDA receptor function was also not associated with either an increase in NMDA receptor mRNA or protein expression.Conclusion:As it has been previously shown that PS-1 is critical for normal NMDA receptor function, our data suggest that the improvement of excitatory neurotransmission occurs through the GPI-1046-induced up-regulation of PS-1.


2009 ◽  
Vol 29 (39) ◽  
pp. 12045-12058 ◽  
Author(s):  
H. Yuan ◽  
K. B. Hansen ◽  
K. M. Vance ◽  
K. K. Ogden ◽  
S. F. Traynelis

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