Retrograde transport of [3H]glycine from the cochlear nucleus to the superior olive in the guinea pig.

The origins of descending glycinergic projections to the guinea pig cochlear nucleus were investigated using retrograde labelling techniques. To identify the cell groups that provide descending projections to the cochlear nucleus, horseradish peroxidase, a nonspecific retrograde neuronal marker, was injected into the cochlear nucleus. After 24 or 48 hours, labelled cell bodies were evident bilaterally in all of the periolivary nuclei that surround the lateral and medial superior olive. The largest numbers of labelled neurons were located in the ventral nucleus of the trapezoid body bilaterally and in the lateral nucleus of the trapezoid body and dorsal periolivary nucleus ipsilaterally. Labelled cells were also present in the inferior colliculus bilaterally and in the contralateral cochlear nucleus. [3H]Glycine was employed as a retrograde tracer to identify the cell groups providing descending glycinergic projections to the cochlear nucleus. Three to 48 hours after injection of 19, 190, or 380 microM [3H]glycine into the cochlear nucleus, retrogradely labelled cell bodies were observed ipsilaterally in all of the periolivary nuclei. No labelled neurons were found in the inferior colliculus. After injections of the highest concentration of [3H]glycine, labelled cells were also found contralaterally in the ventral and lateral nuclei of the trapezoid body and also in the contralateral cochlear nucleus. We conclude that descending glycinergic projections to the cochlear nucleus originate mostly in ipsilateral periolivary cell groups. Minor glycinergic projections originate from the contralateral cochlear nucleus and also from the contralateral ventral and lateral nuclei of the trapezoid body.

Uptake and retrograde transport of [3H]GABA from the cochlear nucleus to the superior olive in the guinea pig.

The purpose of the present study is to determine which descending projections to the cochlear nucleus may use gamma-aminobutyric acid (GABA) as a neurotransmitter. [3H]GABA (120 microM) was injected into the cochlear nucleus of albino and pigmented guinea pigs. After survival times between 0.25 and 16 h, the brain stems were prepared for light microscopic autoradiography. After 2 h survival there was a pulse of label, which progressed through the fibres from the cochlear nucleus to the ipsilateral superior olive. After 5 h, retrogradely labelled neuronal cell bodies and fibres were located in the superior olivary complex bilaterally. In the trapezoid body, clusters of labelled cells were seen in the lateral nucleus, ipsilaterally, and in the ventral nucleus, bilaterally. Also there were labelled cells in the ipsilateral dorsal and anterolateral periolivary nucleus. Large and small cells of several types were labelled. Survival times of 10 h or more resulted in very light, diffuse labelling. Projections to the cochlear nucleus labelled by retrograde transport of horseradish peroxidase that did not take up [3H]GABA included the inferior colliculus, bilaterally, and the cochlear nucleus and periolivary nuclei (other than ventral trapezoid nucleus), contralaterally. The selective labelling of cell groups in the superior olive with the moderately low concentration of [3H]GABA used is consistent with the high-affinity uptake of [3H]GABA by synaptic endings in the cochlear nucleus and its retrograde by transport GABA-ergic neurons. This provides evidence for a descending projection system for inhibitory feedback from the superior olive to the cochlear nucleus.

Long-term degeneration in the cochlear nerve and cochlear nucleus of the adult chinchilla following acoustic overstimulation.

Adult chinchillas were exposed once to an octave-band noise, centered at 4 kHz, and allowed to survive for 16 days or for 1, 2, 4, and 8 months. Axonal degeneration was mapped in the cochlear nucleus, using the Nauta-Rasmussen silver method, and related to hair cell damage and to loss of myelinated nerve fibers in the osseous spiral lamina of the cochlea. Axonal degeneration in the dorsal cochlear nucleus had already reached a peak by 16 days and disappeared after 1 month. Meanwhile, myelinated nerve fiber degeneration in the cochlea extended basally, followed 2 weeks to 2 months later by spread of axonal degeneration into the corresponding high-frequency region of the ventral cochlear nucleus. Axonal degeneration occurred early in the low-frequency region of the ventral cochlear nucleus, followed 2-4 weeks later by spread of myelinated fiber degeneration into more apical regions of the cochlea. New degeneration of axons in the cochlear nerve and in the ventral cochlear nucleus continued to occur for up to 8 months after stimulation. These findings imply that plastic changes in the central auditory pathways could play a role in the long-term effects of cochlear damage and acoustic overstimulation, possibly leading to a chronic neurodegenerative condition in the ear and in the brain.

Quantification of a neurotrophin receptor from submilligram quantities of brain tissue using western blotting.

The immunodensity of the trkB neurotrophin receptor was quantified from submilligram quantities of brain tissue, using approximately 500 microgram samples dissected from the cochlear nucleus (CN) of adult guinea pigs. Tissue samples were hand-homogenized in a lysis buffer. After complete lysis, an aliquot of the lysate was taken to measure total protein, and the remainder was denatured. Proteins in the denatured aliquot were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretically transferred onto an immoblin-P membrane. The membrane was exposed to rabbit anti-trkB and then to anti-rabbit IgG HRP conjugate. The immuno-complex on the membrane was detected by chemiluminescence, which was recorded on autoradiographic film. Autoradiographs were scanned into a computer and the trkB immunobands were quantified by densitometry. This procedure allowed the quantitative comparison of trkB neurotrophin receptor immunodensities between tissue samples. The procedure can be applied to the analysis of small samples of tissue collected from different brain regions or collected from the same region at different times, such as during development or aging, after administering a drug, or after placing a lesion.

Amino acid uptake and release in the guinea pig cochlear nucleus after inferior colliculus ablation.

This study attempts to determine if the neurons in the guinea pig inferior colliculus that project to the cochlear nucleus could use certain amino acid transmitters. The left inferior colliculus was ablated surgically to destroy projections descending to the cochlear nuclei. Nissl and silver stained sections of the brain stem indicated that this procedure destroyed most of the left inferior colliculus, but spared a small amount of collicular tissue ventrally and rostrally. Six to seven days after the ablation, degenerated fibers were present in the right inferior colliculus, in the left lateral lemniscus, and in the cochlear nucleus, bilaterally. Three to five days after the ablation, the uptake and electrically-evoked release of exogenous, radiolabeled D-aspartate, gamma-aminobutyrate, and glycine were measured in the three major subdivisions of the cochlear nucleus, the anteroventral, posteroventral, and dorsal divisions. These activities were compared to those in unlesioned controls. The ablation did not alter the uptake and release of the amino acids in the dorsal and posteroventral divisions of the cochlear nucleus. However, it lowered slightly (by 10-18%) the uptake and release of gamma-aminobutyrate and glycine in the anteroventral division, although the difference from the control group was not statistically significant. These findings suggest that most of the neurons in the inferior colliculus that project to the cochlear nucleus probably do not use glutamate, aspartate, gamma-aminobutyrate, or glycine as a transmitter. However, the possibility remains that a small proportion of the collicular projections to the anteroventral cochlear nucleus might use gamma-aminobutyrate or glycine as a transmitter.

Altered glycinergic synaptic activities in guinea pig brain stem auditory nuclei after unilateral cochlear ablation.

This paper reviews efforts to determine if a unilateral hearing loss altered inhibitory glycinergic synapses in the cochlear nucleus (CN) and the superior olive. In young adult guinea pigs, 2-147 days after unilateral cochlear ablation, we quantified the electrically evoked release and the high-affinity uptake of [(14)C]glycine as measures of transmitter release from glycinergic presynaptic endings and glycine removal from extracellular spaces. The specific binding of [(3)H]strychnine was quantified to measure synaptic glycine receptor activity and/or expression. Three types of post-lesion change were observed. First, several tissues exhibited changes consistent with a persistent deficiency in glycinergic inhibitory transmission. Deficient binding prevailed on the ablated side in the anterior and caudal anteroventral CN, the posteroventral CN and the lateral superior olive (LSO), while glycine release was near normal and uptake was elevated (except in the LSO). However, deficient release prevailed in the dorsal CN, bilaterally, and was accompanied by elevated uptake. Second, the LSO on the intact side exhibited changes consistent with strengthened glycinergic inhibition, as binding was elevated while release and uptake were near normal. Third, several tissues exhibited various transient changes in activity. These types of post-lesion change might contribute to altered auditory functions, which often accompany hearing loss.

Transganglionic transport of D-aspartate from cochlear nucleus to cochlea--a quantitative autoradiographic study.

This study concerns the connections of the inner and outer hair cells and the different types of ganglion cells of the mammalian cochlea with the central nervous system by making use of their putative neurotransmitters. D-[3H]Aspartate (D-ASP), a putative marker for glutamatergic neurons, was injected into the cochlear nucleus of cats and guinea pigs and the cochleas prepared for light microscopic autoradiography after varying survival times. A quantitative, statistical autoradiographic method is described. Grain counts per unit area were made for each of 14 tissue compartments in the cochlea and normalized to permit comparisons between cases. An operationally defined background labeling level was computed for each case and a statistical test for significance applied to the neuron-containing tissue compartments. With increasing survival times, significant labeling appeared successively in the cochlear nerve root, in each type of spiral ganglion cell, and in the neuron-containing tissue compartments of the organ of Corti. The findings are consistent with uptake of D-ASP and retrograde transport by cochlear nerve axons from the cochlear nucleus to the perikarya and peripheral processes of the spiral ganglion. We conclude that axons of all spiral ganglion cells project to the cochlear nucleus and that this nucleus is directly connected with both the inner and outer hair cells. Transganglionic transport of D-ASP from the cochlear nucleus is consistent with the hypothesis that the cochlear nerve axons use glutamate or aspartate as a neurotransmitter.

Evoked release from guinea pig cerebral cortex slices of endogenous 14C-labelled amino acids, labelled via D-[U-14C]glucose.

Release of endogenous amino acids labelled via D-[U-14C]glucose was compared with that of several exogenous labelled amino acids using slices of guinea pig cerebral cortex. Electrical field stimulation evoked a selective release of endogenous [14C]glutamate, [14C]aspartate, and gamma-amino[14C]butyrate (14C-labelled GABA). The selectivity of release correlated well with 14C incorporation into endogenous amino acids. Calculations of the fraction of the tissue radioactivity released indicated that the selectivity was not an artifact due to differential incorporation. Because glucose in mammalian brain is metabolized almost entirely by the so-called 'large compartment', it is tentatively concluded that the releasable 'transmitter pool' of glutamate, aspartate, and GABA is located in this 'large compartment'.

Ca2+-dependence of the evoked release from guinea pig cerebral cortex slices of endogenous 14C-labelled amino acids, labelled via D-[U-14C]glucose.

Spontaneous and electrically evoked release of exogenous labelled amino acids and endogenous amino acids labelled from D-[U-14C]glucose were compared in control and Ca2+-free medium using guinea pig cerebral cortex slices. Spontaneous release of all labelled amino acids, except that of endogenous 14C-labelled threonine-serine-glutamine (unseparated) and exogenous [14C]aspartate, was doubled in Ca2+-free medium. The major portion of the electrically evoked release of endogenous [14C]glutamate, [14C]aspartate, gamma-amino[14C]butyrate (14C-labelled GABA) and exogenous 3H-labelled GABA was Ca2+-inpendent. More than half of the evoked release of the other labelled amino acids was Ca2+-independent. As the pattern of Ca2+-dependence of the evoked release concurred with the selectivity of the evoked release for endogenous [14C]-glutamate, [14C]aspartate, and 14C-labelled GABA, it was concluded that these labelled amino acids were probably released from the amino acid 'transmitter pool'.

Uptake and release of D-aspartate in the guinea pig cochlear nucleus.

This study attempted to determine if L-glutamate (L-Glu) and/or L-aspartate (L-Asp) might be the transmitters of neurons that provide synaptic endings to the cochlear nucleus of the medulla. The uptake and release of D-[3H]aspartate (D-Asp), a putative marker for L-Glu and L-Asp, were measured in the guinea pig cochlear nucleus before and after destruction of the cochlear afferents by cochlear ablation. The cochlear nucleus was dissected into the anteroventral (AVCN), posteroventral (PVCN), and dorsal (DCN) cochlear nuclei. Subdivisions from unlesioned animals took up D-Asp, achieving concentrations in the tissues that were 13-20 times that in the medium. Subsequently, electrical stimulation evoked a Ca2+-dependent release of part of the D-Asp from each subdivision. Disarticulation of the middle ear ossicles, which attenuates acoustic stimulation, produced a modest inhibition of D-Asp release in each subdivision, but did not alter the uptake of D-Asp. Cochlear ablation strongly depressed both the uptake and the release of D-Asp in each subdivision, presumably as a result of destruction of the cochlear nerve endings in the cochlear nucleus. Nevertheless, after lesions, there was a preservation of the uptake and release of D-Asp in the DCN relative to the AVCN and PVCN. These residual activities in the DCN may be mediated by the axonal endings of the granule cells of the cochlear nucleus. The present findings support the hypothesis that the granule cells of the cochlear nucleus, as well as the cochlear nerve fibers, use L-Glu and/or L-Asp as transmitters.

Uptake and release of glycine in the guinea pig cochlear nucleus.

This study attempts to determine if the cochlear nucleus (CN) contains glycinergic synaptic endings. The uptake and release of exogenous radiolabeled glycine were measured in vitro in the three major subdivisions of the guinea pig CN: anteroventral, posteroventral, and dorsal. A kinetic analysis of [3H]glycine uptake revealed the presence in each CN subdivision of a high- and a low-affinity uptake mechanism. The high-affinity mechanism had a Km of 25.2-30.5 microM and a Vmax of 3.8-4.8 nmol/10 mg of cell water/5 min, whereas the low-affinity mechanism had a Km of 633-718 microM and a Vmax of 26.6-37.1 nmol/10 mg of cell water/5 min. At steady state, the high-affinity mechanism accumulated 10 microM [3H]glycine from the medium, achieving tissue concentrations that were 13-24 times that in the medium. The high-affinity uptake was dependent on the temperature and on the concentrations of NaCl and glucose in the incubation medium. It exhibited a high degree of substrate specificity, as determined by the effects of structural analogues of glycine on the uptake of [3H]glycine. Each CN subdivision also contained two mechanisms mediating [14C]glycine release. One was activated by depolarizing electrical stimuli, produced a rapid transient release of [14C]glycine, and was dependent on the presence of extracellular Ca2+. The other was continuous, producing a slow spontaneous efflux of [14C]glycine. Released glycine could be removed primarily by uptake, because during release measurements, the amount of [14C]glycine detected in the medium decreased when glycine uptake activity was optimized. The electrically evoked, Ca2+-dependent release and the high-affinity uptake of glycine may mediate the synaptic release and inactivation of glycine, respectively. These findings, therefore, support the presence of glycinergic synaptic endings in each CN subdivision.

Uptake and release of glycine in the guinea pig cochlear nucleus after axotomy of afferent or centrifugal fibers.

Glycine may be an inhibitory transmitter in the mammalian cochlear nucleus (CN). This study attempts to determine if cochlear and/or centrifugal projections to the CN use glycine as a transmitter. The high-affinity uptake and electrically evoked release of exogenous [14C]glycine were measured in vitro in the three major subdivisions of the guinea pig CN: the anteroventral, posteroventral, and dorsal cochlear nuclei (AVCN, PVCN, and DCN, respectively). [14C]Glycine (3.4 microM) was taken up by each subdivision, reaching tissue concentrations six to seven times that in the medium. Subsequent electrical stimulation evoked a Ca2+-dependent release of [14C]glycine from each subdivision. These activities were compared in subdivisions fr0m unlesioned animals, and from animals with lesions of centrifugal or cochlear projections to the CN. Two knife-cut lesions were made to interrupt centrifugal projections to the CN lying in the right acoustic striae and trapezoid body. In one group of animals, centrifugal fibers projecting mainly to the right AVCN and PVCN were severed, which reduced [14C]glycine uptake and release by 44-53% in these subdivisions, but not in the right DCN. In another group of animals, fibers projecting mainly to the right PVCN and DCN were severed, which reduced [14C]glycine uptake and release by 33-47% in these subdivisions, but not in the right AVCN. In CN subdivisions contralateral to either lesion there was no significant change in [14C]glycine uptake or release. Neither of these lesions altered the uptake or release of D-[3H]aspartate in the right or the left CN. Ablation of the left cochlea, which presumably destroyed cochlear nerve fibers unilaterally, had no effect on [14C]glycine uptake and release. These observations suggest that centrifugal projections contribute a proportion of the glycinergic synaptic endings in the CN. In addition, some glycinergic endings probably arise from neurons intrinsic to the CN. The cochlear nerve contains very few, if any, glycinergic fibers.

Amino acid levels in the guinea pig spinal gray matter after axotomy of primary sensory and descending tracts.

This study attempts to determine if the axonal endings of dorsal root sensory fibers and of descending axons to the spinal gray matter in the guinea pig store glutamate and/or aspartate. Bilateral dorsal rhizotomy (spinal segments C5-T1) and partial cordotomy (segment C5, right side) were used to interrupt primary sensory and descending tracts, respectively. At 1 and 2 days after surgery, amino acid levels were determined in regions microdissected from areas of the gray matter of spinal segment C7 that receive heavy projections from the primary sensory and the descending tracts. These regions were identified by visualizing the degeneration of axons and their terminal fields in silver-impregnated light microscopic preparations of the spinal cord. After dorsal rhizotomy, the heaviest degeneration in the spinal gray appeared centrally in laminae II-IV and medially in laminae IV-VI. The levels of aspartate, glutamate, and gamma-aminobutyrate were reduced by 34, 21, and 26% in laminae II-IV and 28, 33, and 23% in medial laminae IV-VI. The levels of glycine, alanine, and threonine-serine-glutamine (unseparated) were increased. After partial cordotomy, the heaviest degeneration in the spinal gray appeared laterally in laminae IV-VI, dorsolaterally in lamina VII, and in lamina IX. The levels of aspartate and glutamate were reduced by 22 and 28% in lateral laminae IV-VI and by 26 and 28% in dorsolateral laminae VII and IX. Glycine levels were reduced by 9% in dorsolateral laminae VII and IX. The levels of gamma-aminobutyrate, alanine, and threonine-serine-glutamine were either unchanged or raised. These findings suggest that the axonal endings of the primary sensory and of one or more of the descending tracts probably contain relatively high levels of glutamate and aspartate, and that they may use these amino acids as transmitters. The partial deafferentation of spinal interneurons and the destruction of some propriospinal fibers probably caused the losses of gamma-aminobutyrate and glycine, and contributed modestly to those of aspartate.

Evidence for a glutamatergic pathway from the guinea pig auditory cortex to the inferior colliculus.

We attempt to provide evidence that the projection from the guinea pig auditory cortex (AC) to the inferior colliculus (IC) may contain glutamatergic or GABAergic fibers. Seven days after unilateral AC aspiration, histological studies indicated almost complete AC destruction and preterminal degeneration of fibers and terminal fields in the dorsal cortex (DCIC), external cortex (ECIC), and central nucleus (CNIC) of the IC ipsilateral to the ablated AC. Contralaterally, degeneration appeared in the DCIC. AC ablation depressed the electrically evoked Ca(2+)-dependent release of D-[3H]aspartate (D-[3H]Asp) in the ipsilateral DCIC, ECIC, and CNIC, and D-[3H]Asp uptake in the CNIC. Together with other evidence that the corticocollicular pathway is excitatory, these findings suggest that this projection may contain glutamatergic and/or aspartatergic (Glu/Asp-ergic) fibers. Glutamic acid decarboxylase immunoreactivity was not apparent in presumed pyramidal cells of layer V of the AC retrogradely labeled with biotinylated dextran injected into the ipsilateral IC. Thus, corticocollicular neurons probably do not synthesize GABA and may not be GABAergic. However, AC ablation depressed [14C]GABA release from the ipsilateral DCIC and ECIC, and [14C]GABA uptake in the DCIC. These findings are consistent with the atrophy or down-regulation of some subcortical neurons that mediate GABAergic transmission in the IC.

Decreased uptake and release of D-aspartate in the guinea pig spinal cord after dorsal root section.

This study attempts to determine if L-glutamate and/or L-aspartate may be transmitters of dorsal sensory neurons. The uptake and the electrically evoked release of D-[3H]aspartate, a putative marker for L-glutamate and L-aspartate, were measured in the cervical enlargement (segments C4-T1) of the guinea pig spinal cord before and after cutting dorsal roots C5-T1 on the right side. The uptake and the release of gamma-aminobutyric acid (GABA) also were measured as indices of the integrity of GABAergic neurons in the spinal cord. The cervical enlargement was excised and divided into left and right halves, then into dorsal and ventral quadrants. Quadrants from unlesioned animals took up D-aspartate and GABA, achieving concentrations in the tissues which were 14-25 times that in the medium. Subsequently, electrical stimulation evoked a Ca2+-dependent release of D-aspartate and of GABA. The uptake and release of D-aspartate and GABA were similar in tissues taken from intact and sham-operated animals. However, dorsal rhizotomy, without damage to dorsal radicular or spinal blood vessels, depressed the uptake (by 22-29%) and the release (by 50%) of D-aspartate only in quadrants ipsilateral to the lesion. The uptake and the release of GABA were unchanged. In transverse sections of the cervical enlargement, stained to reveal degenerating fibers, by far the heaviest loss of axons occurred in the cuneate fasciculus and in the gray matter ipsilateral to the cut dorsal roots. These findings suggest that the synaptic endings of dorsal sensory neurons probably mediate the uptake and the release of D-aspartate and, therefore, may use L-glutamate or L-aspartate as a transmitter. When spinal blood vessels were damaged during dorsal rhizotomy, the deficits in D-aspartate uptake and release were larger than those in the absence of vascular damage and were accompanied by deficits in GABA uptake and release. These findings imply that vascular damage results in the loss of intraspinal neurons, some of which probably mediate the uptake and release of D-aspartate and, therefore, may use L-glutamate and/or L-aspartate as a transmitter.

Kainate-enhanced release of D-[3H]aspartate from cerebral cortex and striatum: reversal by baclofen and pentobarbital.

A study was made of the actions of the excitant neurotoxin, kainic acid, on the uptake and the release of D-[2,3-3H]aspartate (D-ASP) in slices of guinea pig cerebral neocortex and striatum. The slices took up D-ASP, reaching concentrations of the amino acid in the tissue which were 14-23 times that in the medium. Subsequently, electrical stimulation of the slices evoked a Ca2+-dependent release of a portion of the D-ASP. Kainic acid (10(-5)-10(-3) M) produced a dose-dependent inhibition of D-ASP uptake. The electrically evoked release of D-ASP was increased 1.6-2.0 fold by 10(-5) and 10(-4)M kainic acid. The kainate-enlarged release was Ca2+-dependent. Dihydrokainic acid, an analogue of kainic acid with little excitatory or toxic action, did not increase D-ASP release but depressed D-ASP uptake. Attempts were made to block the action of kainic acid with baclofen and pentobarbital, compounds which depress the electrically evoked release of L-glutamate (L-GLU) and L-aspartate (L-ASP). Baclofen (4 X 10(-6)M), an antispastic drug, and pentobarbital (10(-4)M), an anesthetic agent, each inhibited the electrically evoked release of D-ASP and prevented the enhancement of the release above control levels usually produced by 10(-4)M kainic acid. It is proposed that 10(-5) and 10(-4)M kainic acid may enhance the synaptic release of L-GLU and L-ASP from neurons which use these amino acids as transmitters. This action is prevented by baclofen and pentobarbital. In view of the possibility that cell death in Huntington's disease could involve excessive depolarization of striatal and other cells by glutamate, baclofen might be effective in delaying the loss of neurons associated with this condition.

Decreased uptake and release of D-aspartate in the guinea pig spinal cord after partial cordotomy.

This study attempts to determine if L-glutamate and/or L-aspartate may be transmitters of neural tracts descending from the brain to the spinal cord. The uptake and electrically evoked release of D-[3H]aspartate, a putative marker for L-glutamate and L-aspartate, were measured in the cervical enlargement of the guinea pig spinal cord. These activities were compared using unlesioned animals and others with a lesion on the right side of the spinal cord. Partial cordotomy (segment C5) produced a heavy loss of descending fibers, a small loss of primary sensory fibers, and a depression of the uptake and the Ca2+ -dependent, electrically evoked release of D-aspartate ipsilateral and caudal to the lesion. Contralaterally, there was a moderate loss of corticospinal fibers, some loss of other descending axons, and a depression of D-aspartate release. Dorsal rhizotomy (segments C4-T1) produced a heavy loss of primary sensory fibers ipsilateral to the lesion. Ipsilaterally, but not contralaterally, the uptake and release of D-aspartate were depressed. Degeneration after partial cordotomy in combination with dorsal rhizotomy was assumed to be the sum of that produced by each lesion separately. This combined lesion depressed D-aspartate uptake ipsilaterally and depressed D-aspartate release on both sides of the cervical enlargement. None of the lesions altered the uptake and the evoked release of [3H]GABA. These findings support the hypothesis that the synaptic endings of one or more neural tracts descending from the brain to the spinal cord mediate the uptake and release of D-aspartate and, therefore, may use L-glutamate or L-aspartate as a transmitter.