Differential cellular and subcellular localization of ampa receptor-binding protein and glutamate receptor-interacting protein.
Excitatory synaptic currents in the mammalian brain are typically mediated by the neurotransmitter glutamate, acting at AMPA receptors. We used immunocytochemistry to investigate the distribution of AMPA receptor-binding protein (ABP) in the cerebral neocortex. ABP was most prominent in pyramidal neurons, although it was also present (at lower levels) in interneurons. ABP and its putative binding partners, the GluR2/3 subunits of the AMPA receptor, exhibited prominent cellular colocalization. Under appropriate processing conditions, colocalization could also be documented in puncta, many of which could be recognized as dendritic spines. However, a sizable minority of GluR2/3-positive puncta were immunonegative for ABP. Because glutamate receptor-interacting protein (GRIP) may also anchor GluR2, we studied the relative distribution of ABP and GRIP. There was extensive colocalization of these two antigens at the cellular level, although GRIP, unlike ABP, was strongest in nonpyramidal neurons. Different parts of a single dendrite could stain selectively for ABP or GRIP. To further characterize this heterogeneity, we investigated punctate staining of neuropil using synaptophysin and the membrane tracer DiA to identify probable synapses. Some puncta were comparably positive for both ABP and GRIP, but the majority were strongly positive for one antigen and only weakly positive or immunonegative for the other. This heterogeneity could be seen even within adjacent spines of a single dendrite. These data suggest that ABP may act as a scaffold for AMPA receptors either in concert with or independently from GRIP.
Burette
A
A
Department of Cell Biology and Anatomy, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Khatri
L
L
Wyszynski
M
M
Sheng
M
M
Ziff
E B
EB
Weinberg
R J
RJ
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D000075202:Contraindications; D003712:Dendrites / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins; D007395:Interneurons / Q000166:cytology / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D019579:Neocortex / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019581:Neuropil / Q000378:metabolism / Q000648:ultrastructure; D009928:Organ Specificity; D017966:Pyramidal Cells / Q000166:cytology / Q000378:metabolism; D011726:Pyridinium Compounds; D051381:Rats; D017207:Rats, Sprague-Dawley; D013569:Synapses / Q000378:metabolism; D016708:Synaptophysin / Q000378:metabolism
D002352:Carrier Proteins; C491308:Grip1 protein, rat; C118486:Grip2 protein, rat; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D011726:Pyridinium Compounds; D016708:Synaptophysin; C076868:4-(4-dihexadecylaminostyryl)-N-methylpyridium