Postsynaptic activation at the squid giant synapse by photolytic release of L-glutamate from a 'caged' L-glutamate.
1. Pharmacological evidence suggests L-glutamate is a strong candidate as a transmitter at the giant synapse of the squid. Postsynaptic activation at the giant synapse cannot be effected by conventional application of putative neurotransmitters by iontophoresis or perfusion, apparently because the complex structure of the synapse prevents a sufficiently rapid change in concentration at the postsynaptic membrane. Flash photolytic release of L-glutamate from a pharmacologically inert 'caged' L-glutamate pre-equilibrated in the stellate ganglion of Alloteuthis or Loligo was used to determine whether L-glutamate can produce postsynaptic activation when released rapidly in the synaptic clefts. 2. The preparation, reaction mechanism and properties of the caged L-glutamate, N-1-(2-nitrophenyl)ethoxycarbonyl-L-glutamate, are described. The product quantum yield on photolysis was 0.65 (+/- 0.05). On flash photolysis glutamate release followed a single exponential time-course in the pH range 5.5-7.8. The rate constant was proportional to [H+] and was 93 s-1 at pH 5.5 and 16 degrees C in artificial sea water (ionic strength, I = 0.68 M). 3. At pH 7.8 flash photolysis of caged glutamate pre-equilibrated in the synapse caused only a slow depolarization. A second photolytic release of L-glutamate or transsynaptic activation produced no further depolarization, suggesting desensitization and inactivation of postsynaptic mechanisms by the initial pulse of L-glutamate. 4. Synaptic transmission in the giant synapse was normal at pH 5.5. Flash photolysis at pH 5.5 caused rapid production of L-glutamate within the synaptic cleft and a fast postsynaptic depolarization which generated postsynaptic action potentials.(ABSTRACT TRUNCATED AT 250 WORDS)
Corrie
J E
JE
National Institute for Medical Research, Mill Hill, London.
DeSantis
A
A
Katayama
Y
Y
Khodakhah
K
K
Messenger
J B
JB
Ogden
D C
DC
Trentham
D R
DR
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D049832:Decapodiformes; D005971:Glutamates / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D066298:In Vitro Techniques; D009412:Nerve Fibers / Q000502:physiology; D018377:Neurotransmitter Agents / Q000502:physiology; D010782:Photolysis; D017470:Receptors, Glutamate / Q000502:physiology; D013233:Stellate Ganglion / Q000166:cytology; D013569:Synapses / Q000502:physiology*
D005971:Glutamates; D018377:Neurotransmitter Agents; D017470:Receptors, Glutamate