Viral delivery of NR2D subunits reduces Mg2+ block of NMDA receptor and restores NT-3-induced potentiation of AMPA-kainate responses in maturing rat motoneurons.
N-methyl-D-aspartate (NMDA) responsiveness of motoneurons declines during the initial 2 postnatal weeks due to increasing Mg2+ block of NMDA receptors. Using gene chip analyses, RT-PCR, and immunochemistry, we have shown that the NR2D subunit of the NMDA receptor (NMDAR), known to confer resistance to Mg2+ block, also declines in motoneurons during this period. We injected a viral construct (HSVnr2d) into the lumbar spinal cord on postnatal day 2 in an attempt to restore NMDAR function in motoneurons during the second postnatal week. Following HSVnr2d injection, we detected elevated levels of NR2D mRNA in spinal cord samples and NR2D protein specifically in motoneurons. These molecular changes were associated with marked functional alterations whereby NMDAR-mediated responses in motoneurons associated with both dorsal root (DR) and ventrolateral funiculus (VLF) inputs returned to values observed at E18 due to decreased Mg2+ blockade. Viruses carrying the beta-galactosidase gene did not induce these effects. NT-3 is known to potentiate AMPA-kainate responses in motoneurons if the response has an NMDAR-mediated component and thus is normally ineffective during the second postnatal week. Restoration of NMDAR-mediated responsiveness in the second postnatal week was accompanied by a return of the ability of neurotrophin-3 (NT-3) to potentiate the AMPA-kainate responses produced by both DR and VLF synaptic inputs. We conclude that delivery of the gene for a specific NMDA subunit can restore properties characteristic of younger animals to spinal cord motoneurons. This approach might be useful for enhancing the function of fibers surviving in the damaged spinal cord.
Arvanian
Victor L
VL
Dept. of Neurobiology and Behavior, SUNY at Stony Brook, Life Sciences Bldg. Rm. 550, Stony Brook, NY 11794-5230, USA.
Bowers
William J
WJ
Petruska
Jeffrey C
JC
Motin
Vladimir
V
Manuzon
Honeyleen
H
Narrow
Wade C
WC
Federoff
Howard J
HJ
Mendell
Lorne M
LM
D000818:Animals; D000831:Animals, Newborn; D003423:Critical Period, Psychological; D004594:Electrophysiology; D005822:Genetic Vectors; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology*; D009046:Motor Neurons / Q000187:drug effects*; D020933:Neurotrophin 3 / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000819:agonists*; D018092:Kainic Acid Receptors / Q000819:agonists*; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018139:Simplexvirus / Q000235:genetics; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects / Q000254:growth & development; D013569:Synapses / Q000187:drug effects
D008274:Magnesium; D020933:Neurotrophin 3; D018091:Receptors, AMPA; D018092:Kainic Acid Receptors; D016194:Receptors, N-Methyl-D-Aspartate; C464436:NR2D NMDA receptor