Surface-accessible GABA supports tonic and quantal synaptic transmission.
Exocytosis is commonly viewed as the only secretory process able to account for quantal forms of fast synaptic transmission. However, the demonstrated variability and composite properties of miniature postsynaptic signals are not easily explained by all-or-none exocytotic discharge of transmitter in solution from inside vesicles. Recent studies of endocrine secretion have shown that hormone release does not coincide with exocytosis due to its trapping in the core matrix of the granule. Thus, we tested whether the synaptic transmitter GABA could also be held in a matrix before being released. Using confocal microscopy and flow cytometry of embryonic rat hippocampal neurons, we found a GABA immunoreaction at the surface of live cell bodies and growth cones that coincided spatially and quantitatively with the binding of tetanus toxin fragment C (TTFC). TTFC binds predominantly at membrane sites containing the trisialoglycosphingolipid GT1b. Using flow cytometry, GT1b-containing liposomes preincubated in 100 nM GABA exhibited the same relationship between GABA and TTFC surface binding as found on neurons and growth cones. Embryonic neurons differentiated in culture expressed initially a tonic, and after 3-5 days, transient, postsynaptic signals mediated by GABA acting at GABA(A) receptor/Cl(-) channels. A stream of saline applied to the neuronal surface rapidly and reversibly suppressed both tonic and transient signals. A brief application of the GABAmimetic isoguvacine immediately transformed both tonic and transient GABAergic signals into tonic and transient isoguvacinergic signals. These results and those in the literature are consistent with an immediately releasable compartment of transmitter accessible from the presynaptic surface.
Vautrin
J
J
Laboratory of Neurophysiology, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD 20892, USA. vautrin@codon.nih.gov
Maric
D
D
Sukhareva
M
M
Schaffner
A E
AE
Barker
J L
JL
D000327:Adsorption; D000818:Animals; D002478:Cells, Cultured; D018118:Chloride Channels / Q000378:metabolism; D004594:Electrophysiology; D005089:Exocytosis / Q000502:physiology*; D005260:Female; D005434:Flow Cytometry; D018755:GABA Agonists / Q000494:pharmacology; D005732:Gangliosides / Q000032:analysis; D007539:Isonicotinic Acids / Q000494:pharmacology; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D017729:Presynaptic Terminals / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000032:analysis / Q000378:metabolism*
D018118:Chloride Channels; D018755:GABA Agonists; D005732:Gangliosides; D007539:Isonicotinic Acids; D008081:Liposomes; D008565:Membrane Proteins; D011963:Receptors, GABA-A; D005680:gamma-Aminobutyric Acid; C016481:trisialoganglioside GT1; C015300:isoguvacine