Primary afferent activity, putative excitatory transmitters and extracellular potassium levels in frog spinal cord.
1. Changes in extracellular K+ activity were measured with ion-selective microelectrodes in the grey matter of the isolated hemisected frog spinal cord. The magnitude of the elevation of [K+]o (delta[K+]o) produced by repetitive stimulation (25 Hz, 10 s) of afferent fibres in the sciatic nerve was monotonically related to the strength of the electrical stimuli applied to the sciatic nerve. Repetitive stimulation of the largest diameter A alpha and A beta fibres, which were found histologically to comprise only 11% of the afferent axons in the dorsal root, elevated [K+]o to approximately 60% of the maximum level seen when all afferent fibres were stimulated. 2. Addition of Mg2+ (20 mM) to Ringer solution devoid of Mg2+ reduced delta[K+]o by over 85% suggesting that about 15% of delta[K+]o results from action potentials in presynaptic primary afferents. When 20 mM-Mg2+ was added to spinal cords bathed in Ringer solution containing a physiological (i.e. 1.0 mM) concentration of Mg2+, delta[K+]o was reduced by ca. 65-75% indicating that in spinal cords bathed in medium containing 'physiological' concentrations of Mg2+ about 25-35% of the K+ is released from primary afferent fibres. 3. Application of excitatory amino acids and agonists increased [K+]o with the following potency pattern: quisqualate greater than kainate greater than NMDA (N-methyl-D-aspartate) greater than glutamate greater than aspartate. 4. D(-)-2-Amino-5-phosphonovalerate (APV), an NMDA antagonist, reduced [K+]o by only about 50%, but kynurenate, an NMDA and non-NMDA antagonist, reduced [K+]o by approximately 85%; i.e. the same levels observed when synaptic transmission was blocked with 20 mM-Mg2+. These findings support the idea that synaptic release of excitatory amino acids such as L-glutamate and/or L-aspartate and subsequent activation of specific receptors by these putative transmitters are necessary for the postsynaptic component of delta[K+]o. 5. Addition of tachykinins elevated [K+]o but the effect appeared to require the participation of excitatory amino acids because it was blocked by APV and by kynurenate. 6. The finding that tetrodotoxin substantially reduced the ability of excitatory amino acid agonists and tachykinins to elevate [K+]o suggests that discharges in interneurones as a result of excitatory amino acid receptor activation are responsible for the postsynaptic component of delta[K+]o.
Davidoff
R A
RA
Department of Neurology, University of Miami School of Medicine, FL.
Hackman
J C
JC
Holohean
A M
AM
Vega
J L
JL
Zhang
D X
DX
D000200:Action Potentials / Q000187:drug effects; D000596:Amino Acids / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000818:Animals; D004305:Dose-Response Relationship, Drug; D005110:Extracellular Space / Q000502:physiology*; D066298:In Vitro Techniques; D007705:Kinins / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D009412:Nerve Fibers / Q000502:physiology*; D009475:Neurons, Afferent / Q000502:physiology*; D011188:Potassium / Q000502:physiology*; D011894:Rana pipiens; D013116:Spinal Cord / Q000502:physiology*; D013126:Spinal Nerve Roots / Q000502:physiology; D013779:Tetrodotoxin / Q000494:pharmacology
D000596:Amino Acids; D007705:Kinins; D013779:Tetrodotoxin; D008274:Magnesium; D011188:Potassium