Lateral mobility and anchoring of recombinant GABAA receptors depend on subunit composition.
The clustering of type A gamma-aminobutyric acid receptors (GABA(A)R) at discrete and functionally significant domains on the nerve cell surface is an important determinant in the integration of synaptic inputs. To discern the role that the subunits of the GABA(A)R play in determining the receptor's cell surface topography and mobility, the alpha1, beta1, beta3, and gamma2s subunits were transfected into COS7, HEK293, and PC12 cells and the distribution and cell surface mobility of these recombinant receptors were examined. Our results show that alpha1 subunits are retained in the endoplasmic reticulum while beta1 and beta3 subunits are sorted to the plasma membrane where they form clusters. Co-expression and co-assembly of alpha1 and beta3 subunits result in the rescue of intracellular alpha1 subunits, which are transported as alphabeta subunit complexes to the cell surface where they formed clusters. Fluorescence photobleach recovery and single particle tracking of recombinant receptors show that, despite clustering, beta3 subunit homooligomers are mobile within a cell surface domain. Inclusion of alpha1 in beta3 or beta3gamma2s complexes, however, dramatically reduces the receptor's lateral mobility in COS 7 and PC12 cells and anchors GABA(A)Rs on the cell surface, suggesting the formation of a direct link to a component of the cytoskeleton. The mobility of recombinant receptors that include the alpha1 subunit mirrors the mobility of GABA(A)Rs on cell bodies and dendrites of cortical and spinal cord neurons. The results suggest that incorporation of alpha1 subunits give rise to a population of GABA(A)Rs that are immobilized on the cell surface.
Peran
M
M
Departamento de Bioquimica, Facultad de Medicina, Universidad de Malaga, Malaga, Spain.
Hicks
B W
BW
Peterson
N L
NL
Hooper
H
H
Salas
R
R
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology* / Q000648:ultrastructure; D003599:Cytoskeleton / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000502:physiology; D006801:Humans; D009433:Neural Inhibition / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D016716:PC12 Cells; D021122:Protein Subunits / Q000235:genetics*; D021381:Protein Transport / Q000502:physiology; D051381:Rats; D011940:Receptor Aggregation / Q000235:genetics*; D011963:Receptors, GABA-A / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D009435:Synaptic Transmission / Q000502:physiology; D014162:Transfection; D005680:gamma-Aminobutyric Acid / Q000502:physiology
D021122:Protein Subunits; D011963:Receptors, GABA-A; D011993:Recombinant Fusion Proteins; D005680:gamma-Aminobutyric Acid