Primary afferent terminals in spinal cord express presynaptic AMPA receptors.
Larger dorsal root ganglion neurons are stained by an antibody for the C terminus of glutamate receptor subunit 2 (GluR2) and GluR3 (GluR2/3) rather than by an antibody for GluR4. In dorsal roots, anti-GluR2/3 stains predominantly myelinated fibers; anti-GluR4 or anti-GluR2/4 stains predominantly unmyelinated fibers. In the dorsal horn, puncta immunopositive for synaptophysin and GluR2/3 are predominantly in laminas III and IV, whereas puncta immunopositive for synaptophysin and GluR4 or GluR2/4 are predominantly in laminas I and II. Puncta immunopositive for GluR2/3 costain with the B subunit of cholera toxin, whereas puncta immunopositive for GluR2/4 costain with isolectin B4 after injections of these tracers in the sciatic nerve. No puncta costain with calcitonin gene-related peptide and AMPA receptor subunits. Electron microscopy indicates that AMPA receptor-immunopositive terminals are more numerous than suggested by confocal microscopy. Of all synapses in which immunostaining is presynaptic, postsynaptic, or both, the percentage of presynaptic immunostain is approximately 70% with anti-GluR4 or anti-GluR2/4 (in laminas I-III), 25-30% with anti-GluR2/3 (in laminas III and IV), and 5% with anti-GluR2 (in laminas I-III). Because of fixation constraints, the types of immunostained terminals could be identified only on the basis of morphological characteristics. Many terminals immunostained for GluR2/3, GluR4, or GluR2/4 have morphological features of endings of primary afferents. Terminals with morphological characteristics of presumed GABAergic terminals are also immunostained with anti-GluR2/4 and anti-GluR4 in laminas I and II and with anti-GluR2/3 in laminas III and IV. The conspicuous and selective expression of presynaptic AMPA receptor subunits may contribute to the characteristic physiological profile of different classes of primary afferents and suggests an important mechanism for the modulation of transmitter release by terminals of both myelinated and unmyelinated primary afferents.
Lu
Chun-Rong
CR
Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA. chunrong_lu@med.unc.edu
Hwang
Se Jin
SJ
Phend
Kristen D
KD
Rustioni
Aldo
A
Valtschanoff
Juli G
JG
D000818:Animals; D005455:Fluorescent Antibody Technique; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D009413:Nerve Fibers, Myelinated / Q000378:metabolism / Q000648:ultrastructure; D009475:Neurons, Afferent / Q000378:metabolism* / Q000648:ultrastructure; D020671:Posterior Horn Cells / Q000378:metabolism / Q000648:ultrastructure; D017729:Presynaptic Terminals / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000096:biosynthesis*; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism*; D016707:Tissue Fixation / Q000379:methods
D018091:Receptors, AMPA; C104723:glutamate receptor ionotropic, AMPA 3; C104724:glutamate receptor ionotropic, AMPA 4; C104722:glutamate receptor ionotropic, AMPA 2