Michael R. Martin

Structure-activity relations of excitatory amino acids on frog and rat spinal neurones.

1 A series of compounds structurally related to glutamic acid has been tested on frog and rat spinal neurones. The substances were added to procaine‐containing medium bathing the isolated hemisected spinal cord of the frog, and their potencies in depolarizing motoneurones were assessed by the magnitude of the potential produced in the ventral root. The electrophoretic technique was used to administer the substances around single interneurones of the rat spinal cord and the relative potencies of the compounds as excitants assessed by the magnitude of the currents required to produce similar rates of neuronal firing. 2 Parallel structure‐activity relations were observed in the two series of experiments, suggesting that the receptors for excitatory amino acids on frog and rat spinal neurones are similar. 3 Quisqualate, domoate and kainate were the strongest excitants in both animals, with potencies around two orders of magnitude higher than that of l‐glutamate. 4 2,4,5‐Trihydroxyphenylalanine (6‐OH‐DOPA) was a stronger excitant and l‐3,4‐dihydroxyphenylalanine (l‐DOPA) a weaker excitant than l‐glutamate on frog spinal motoneurones. The former compound was also a more potent convulsant than l‐glutamate on intraventricular injection into mouse brain. The lack of activity of 6‐OH‐DOPA on electrophoretic administration was attributed to oxidation. 5 Unlike the majority of amino acid excitants, several of the compounds shown in the present work to have moderate excitatory activity are not anionic at physiological pH. This indicates either that two negatively charged groups are not essential for interaction with a common excitatory receptor, or that more than one type of receptor is involved in the actions demonstrated.

Morphology of the facial nucleus of the rat.

The facial nucleus of the rat can be divided into 5 morphological subdivisions. Using a method for the correlation of the observed subdivisions with antidromic field profiles, the orgins of the major muscle branches of the facial nerve in the motor nucleus were determined. The posterior auricular branch is in the medial, the cervical in the ventromedial, the inferior and superior buccolabiales in the lateral, the zygomatic in the dorsal and the temporal and digastric in the intermediate subdivision. The results are consistent with an organization of the motor nucleus reflecting a corresponding topographic organization of the facial musculature.

The seventh cranial nerve of the rat. Visualization of efferent and afferent pathways by cobalt precipitation

The cobalt sulphide precipitation technique, in conjunction with Timms intensification procedure, was used to delineate the afferent and efferent intramedullary pathways of the seventh cranial nerve complex in the rat. The branchial motor nucleus with the accompanying first part, genu, and second part of the root are described. The motor branches to the superficial facial musculature do not contain fibres of geniculate ganglion origin or fibres which terminate in the spinal trigeminal nucleus. The motor branches to the deep facial muscles arise from the dorsal part of the branchial motor nucleus and traverse to the midline medial to the genu, then project under the genu into the lateral reticular formation before exiting with the facial nerve. The salivatory and lacrimal nuclei and their intramedullary pathways are described. Sensory fibres from the cutaneous auricular branch enter the spinal trigeminal tract and most of the chorda tympani gustatory fibres enter the fasciculus solitarius. A smaller number of gustatory fibres extend medially to the region of the salivatory nucleus. Fibres of greater superficial petrosal origin also enter the fasciculus solitarius as well as the medial reticular formation. These findings are discussed in relation to previous anatomical, physiological and clinical reports.

Electrophysiological observations on the spinal cord of the normal and dystrophic mouse

A method of carrying out electrophysiological experiments on the mouse spinal cord is described. The conduction velocity in the spinal dorsal roots (DR) of the normal mouse was in the range 10-100 m sec-1 and in the ventral roots (VR) 50-70 m sec-1. In the dystrophic mutant (129 ReJ dy/dy) the conduction velocity for both roots was usually in the range 0.1-2.0 m sec-1. Reflexes from DR to VR were recorded in both mutant and wild type animals and it was concluded that the delays in the mutant reflex were probably due to the slower conduction velocity in the roots. Postsynaptic inhibition and presynaptic inhibition were demonstrated and records were made from Renshaw cells and intracellularly from motoneurones. Delayed activity in spinal reflex paths, probably of supraspinal origin, was more pronounced in the dystrophic mutant. It is concluded that if the dystrophic mouse mutant were to be regarded as a model for human disease then similar reflex delays should be demonstrated in human subjects with muscular dystrophy.

The sensitivity of rat spinal interneurones and Renshaw cells to L-glutamate and L-aspartate

SummaryL-Glutamate and L-aspartate were administered electrophoretically near spinal interneurones and Renshaw cells of pentobarbitone-anaesthetized rats. Other spinal interneurones were consistently more sensitive to L-glutamate than to L-aspartate. Renshaw cells, however, showed no consistent difference in their sensitivity to these two amino acids. The results, which are compared with those reported previously in the cat, support the hypothesis that L-glutamate could be a transmitter at spinal primary afferent terminals.

Effects ofd,l-α-aminoadipate on postsynaptic amino acid responses in cultured mouse spinal cord neurons

Abstract The effects of the dicar☐ylic amino acid, dl -α-aminoadipate (DLAA) on amino acid responses have been investigated using intracellular recordings from mouse spinal cord neurons grown in dissociated cell culture. dl -α-Aminoadipate markedly antagonized postsynaptic responses to iontophoretically applied aspartate; antagonism of glutamate was much less prominent. dl -α-Aminoadipate altered the affinity of aspartate for its receptor while having no observed effects on aspartate-receptor cooperativity. No direct effects of DLAA on membrane potentials or passive membrane properties were seen at the currents used for antagonism. Responses to the inhibitory amino acids GABA and glycine were unaffected by DLAA.

Quisqualamine, a novel γ-aminobutyric acid (GABA)-related depressant amino acid

A new substance, quisqualamine, the decarboxylated analogue of quisqualic acid, predictably depressed electrical activity of neurons of the frog and rat spinal cord in vitro and of the mouse spinal cord in vivo. In the in vitro preparations, the action of quisqualamine was associated with a prolonged depolarization of primary afferent terminals which was sensitive to blockade by picrotoxin and bicuculline and which was also depressed by strychnine. This suggests an interaction of quisqualamine with presynaptic receptors for both GABA and β‐alanine. Post‐synaptic actions of quisqualamine, which were less marked than those at pre‐synaptic sites, also appeared to be predominantly GABA‐mimetic in vitro, though a sensitivity to the GABA‐antagonist bicuculline could not be demonstrated in vivo.

Pharmacological studies on facial motoneurones in the rat

Experiments were performed on 39 male albino rats anaesthetized with pentobarbitone sodium and paralyzed with gallamine triethiodide. Facial motoneurones consistently showed a progressive alteration of spike shape and decrease in spike firing frequency during the continuous microelectrophoretic application of the excitant amino acids D,L-homocysteate, L-glutamate, L-aspartate, and kainate, but infrequently during that of N-methyl-D-aspartate. The relative potencies of these excitants on facial motoneurones are reported. The potential usefulness of N-methyl-D-aspartate to produce amino acid-evoked motoneurone action potentials is discussed. The microelectrophoretically-applied depressant amino acid antagonist strychnine selectively and reversibly blocked the depressant effects of glycine on facial motoneurones. The depression of amino acid-induced firing of facial motoneurones by stimuli to the lingual or glossopharyngeal nerves were reversibly antagonized by microelectrophoretically applied strychnine but not by bicuculline.