Distribution of muscimol, QNB, and 5HT binding in the vertebrate diencephalon: a comparative study of eight mammals and three non-mammals.

The distribution of muscimol, quinuclidinyl benzilate (QNB), and serotonin (5HT)-bound receptors in the diencephalon was examined by conventional receptor-binding methods in 11 species of amniotes including 2 reptiles, 1 bird, and 8 mammals, selected mostly on the basis of their differing last common ancestor with Anthropoids. We found that receptor binding can help define major subdivisions of the forebrain. The results show that in each of these species, the distribution of muscimol and QNB binding across the four major subdivisions of the diencephalon was consistent; densest in the dorsal thalamus, with hypothalamus and then either ventral thalamus or epithalamus with successively lesser amounts. However, the binding of serotonin (5HT) was most prevalent in the hypothalamus with equivalent amounts in the other diencephalic subdivisions. Myelin- and cell-stained materials showed that the pattern of high-density binding probably is not the secondary result of non-neurochemical factors such as differences in cell or neuropil density or in total available membrane. Perhaps more importantly, the receptor distributions suggest functional roles for major subdivisions across taxa. Results show that GABA-A and muscaranic Ach receptors are common in the dorsal diencephalon across vertebrate species and, therefore, are probably responsible for the gating of information to the cortex. Results show that serotonin is predominant in the hypothalamus. The lack of it in the dorsal thalamus indicates that it is probably not responsible for gating of information to the cortex. Results also show that in nonmammals the amount of GABA-A and muscaranic Ach differs from that found in mammals. For muscaranic Ach, the labeling in marsupials differs from that in placentals. Primates differ from other species (nonmammals and mammals combined) in the amount of 5HT found in the ventral diencephalon and the hypothalamus.

Role of acoustic striae in hearing: reflexive responses to elevated sound-sources.

This report is the fourth in a series describing the results of ablation-behavior experiments directed to the ascending output of the cochlear nuclei as it is conducted centrally within the acoustic striae. This fourth report focuses on the unique physiology of the fusiform or 'output' cells of the dorsal cochlear nucleus whose axons course through the dorsal acoustic stria (DAS). Because electrophysiological studies have shown that the cues for sensing the elevation of a sound source would seem to be best analyzed by the dorsal cochlear nucleus and projected centrally via its DAS, we tested normal cats and cats deprived of DAS for their ability to orient to elevated sources of broad-band noise. For behavioral testing, we made use of reflexive or unconditioned orienting responses to elevated sound sources using a similar method to one we have used previously for azimuth testing (Thompson GC, Masterton RB. Brainstem auditory pathways involved in reflexive head orientation to sound. J Neurophysiol 1978;41:1183-1202). The results show that cats deprived of their DAS do indeed have a marked deficit in their ability to orient to an elevated sound source. Further behavioral testing indicated that this deficit is not the secondary result of an attentional or peripheral motor deficit. Although the present results do not prove that the reflexive deficit is strictly auditory in nature, the deficit is notable in that it is the only one yet known to result from a lesion of the dorsal cochlear nucleus or its central projections.

Lack of projections from medial geniculate body to suprasylvian cortex in cat: a study with horseradish peroxidase.

Because both electrophysiological and behavioral methods have implicated the suprasylvian cortex of cat in audition, its afferents were studied using retrograde transport of horseradish peroxidase. The bulk-filling method was used to maximize the likelihood that virtually all afferents to the area would be labeled. Despite the vivid retrograde labeling of many thalamic cells with this procedure, no direct auditory projections to the suprasylvian cortex could be found in the thalamus (i.e. in medial geniculate body or in the dorsolateral part of the posterior nucleus). Furthermore, very few cells were labeled in the primary auditory cortex of the nearby ectosylvian gyrus. The source of afferents to the suprasylvian cortex originate mostly from the pulvinar-lateral posterior complex and to a lesser extent from ventral lateral and ventral anterior nuclei of the thalamus.

Role of acoustic striae in hearing: discrimination of sound-source elevation.

After years of systematic experimentation, we finally uncovered one thing the dorsal system contributes to hearing which the ventral system may not -- the mechanism for orienting to an elevated sound source [Sutherland, D.P., Masterton, R.B., Glendenning, K.K. (1998) Behav. Brain Res. in press]. This paper follows up this one positive result on a historical background of uniformly negative results. The focus of this report is on the fusiform cells of the dorsal cochlear nucleus whose axons course through the dorsal acoustic stria (DAS). Because electrophysiological studies have shown that the cues for sensing the elevation of a sound source would seem to be best analyzed by the dorsal cochlear nucleus, we tested, behaviorally, normal cats and cats deprived of their DAS or intermediate acoustic stria, bilaterally or ipsilaterally (with or without their contralateral ear deafened), for their ability to orient to elevated sources of broad-band noise. For behavioral testing, we made use of a conventional shock-avoidance procedure. The results lead to the conclusion that DCN and DAS may play no role in learned elevation discriminations. This result builds on that of another of our papers which suggests that a deficit in reflexive discrimination of elevation is strictly auditory in nature [Sutherland, D.P., Masterton, R.B., Glendenning, K.K. (1998) Behav. Brain Res. in press].

Lack of topography in the ventral nucleus of the lateral lemniscus.

In contrast to the ease of finding tonotopicity in other nuclei, both anatomical and electrophysiological methods have failed to demonstrate a clear and simple tonotopic map within the ventral nucleus of the lateral lemniscus (VLL). The present study was undertaken in cat with the hope that methods not used previously in studies of VLL might succeed in demonstrating an orderliness in its exiting fibers (i.e., efferents) or its incoming fibers (i.e., afferents). Since the same organization of ascending frequencies present in the cochlea is maintained in these fibers as well as in all main auditory nuclei, demonstration of a similar organization of frequencies in VLL would be evidence of the cochleo- or tono-topicity of this nucleus. Using triple injection of 3 different fluorescent dyes in inferior colliculus to study efferents, orderly and tonotopic cell-labeling is found in each of the brainstem auditory nuclei, with the notable exception of VLL. Instead, labeling of cell clusters, each cluster containing a small number of cells, is found randomly distributed throughout VLL in all 3 of its spatial dimensions. Using the 2-deoxyglucose (2-DG) method, during stimulation at 6 different frequencies, afferent orderliness, indeed, tonotopicity is found in all major brainstem auditory nuclei, again with the notable exception of VLL. Rather, each frequency evokes 2-DG label throughout VLL. In agreement with the results based on electrophysiological methods, therefore, the anatomical methods used here also yield no evidence of tonotopicity in VLL. Thus, if there is orderliness in VLL's efferents or afferents, it is based on an auditory dimension incommensurate with frequency.

Comparative morphometry of mammalian central auditory systems: variation in nuclei and form of the ascending system.

The volumes of the ten largest subcortical auditory nuclei were measured individually in a sample of 53 mammals, including 16 Australian and four American marsupials. The nuclear sizes relative to the total of subcortical auditory tissue were normalized and then analyzed individually for statistically reliable deviations. The overall form of the entire system of ten nuclei and two nuclear sub-systems (cochlear nuclei, superior olives) were also analyzed for similarities and notable deviations among the animals. The results show that the absolute size of the auditory system varies more than 139-fold among the 53 mammals (with moles the smallest and humans the largest). Log auditory system volume and log brain weight are closely correlated (r = 0.903, p < 0.0001). Bats, kangaroo rats, marmosa opossums, and Norway rats have the largest auditory systems relative to their brain size, while humans have the smallest by far. The other primates also have auditory system/brain size ratios smaller than the sample average, suggesting that the condition in humans is one result of an expansion of non-auditory brain parts rather than a reduction of the auditory system over geological time. The relative sizes of the ten nuclei are well ordered, with the inferior colliculus the largest nucleus by far and medical superior olive the smallest. Because the size of the superior olives, collectively, is reliably related to the size of anteroventral cochlear nucleus (r = 0.744, p < 0.001), and not to the size of dorsal cochlear nucleus, the interconnectivity of the subcortical auditory system is probably a factor in the size of the nuclei. In its overall form, the subcortical auditory system is highly similar among mammals, with an average correlation across nuclei of 0.923. This high value means that the overall form of the system has been relatively stable over geological time. The animals with least deviation from the average form are ring-tailed possums, bandicoots, and yellow-bellied gliders, all marsupials. Those with the most unusual forms are mice, bats, and kangaroo rats, all placentals.

Role of acoustic striae in hearing: mechanism for enhancement of sound detection in cats.

We report the results of behavioral studies in cats conducted first, to demonstrate the presence of a monaural mechanism for the enhancement of signal to noise; and then to examine the necessity or sufficiency of the acoustic striae for this mechanism. The results show that cats do indeed have a monaural mechanism for enhancing the detection of tones in co-located background noise for noise levels at least as high as 60 dB SPL. The ablation-behavior results show that surgical section of the dorsal (DAS) and most of the intermediate (IAS) striae has no measurable effect on this mechanism. In sharp contrast, even partial section of the trapezoid body results in a profound and permanent deficit and this deficit is not accounted for by hearing loss alone. It is concluded that the ascending and descending fibers in the dorsal and intermediate acoustic striae are neither necessary nor sufficient for enhancing the detection of salient sounds in a noisy environment while the ascending or descending fibers in the ventral acoustic stria are sufficient and probably necessary.

Psychoacoustical contribution of each lateral lemniscus.

Although each lateral lemniscus is required for sound localization in its contralateral hemifield, no auditory function is yet known for the neural activity evoked in the lemniscus ipsilateral to a sound source. In an attempt to assess the role played by the ipsilateral lemniscus, monaural cats were tested on an array of psychoacoustical tasks before and after surgical section of one or the other lateral lemniscus. The results show that the lemniscus contralateral to the remaining intact ear is either necessary or sufficient for 24 of the 26 tests administered. However, the lemniscus ipsilateral to the intact ear is both necessary and sufficient (or alternatively, the contralateral lemniscus makes no obvious contribution) to normal thresholds in two of the tasks: detection of low-frequency tones (< 4 kHz) and detection of low-frequency AM modulation. Because of their projections to the ipsilateral inferior colliculus via the ipsilateral lemniscus, the anatomical substrate of these two unusual tasks is probably the fibers from the MSO and possibly, the LSO, ipsilateral to the intact ear.

Acoustic chiasm V: inhibition and excitation in the ipsilateral and contralateral projections of LSO.

When this series of experiments was begun in 1984, the activity of each lateral superior olive (LSO) in the mammalian hindbrain was known to encode the hemifield of acoustic space containing a sound source. However, the almost random bilaterality of its ascending projections seemed to jumble that identification before reaching the midbrain. At the same time, electrophysiological studies of LSO and its efferent target in the inferior colliculus, along with the strictly contralateral deficits in sound localization resulting from unilateral lesions above the level of the superior olives, indicated that hemifield allegiance was largely maintained (though reversed) at the midbrain. Here we present seven lines of biochemical evidence, some combined with prior ablations, supporting the notion that the anatomical segregation of the ipsilateral and contralateral fibers ascending from the LSO is accompanied by a corresponding segregation of their neurotransmitters: most of the ascending ipsilateral projection is probably glycinergic and, hence, inhibitory in effect, while most of the contralateral projection is probably glutamatergic/aspartergic and, hence, excitatory in effect. Taken together, the inhibitory ipsilateral projections and the excitatory contralateral projections serve to amplify functional contralaterality at the higher levels of the auditory system.

Acoustic chiasm. IV: Eight midbrain decussations of the auditory system in the cat.

Conventional retrograde and orthograde axonal transport tract-tracing techniques were used in cats to explore the auditory decussations and commissures in the upper pons and midbrain. In all, 8 decussations differing either in origin or in contralateral termination were found. Three of the 8 decussations (from the dorsal nucleus of the lateral lemniscus to the contralateral dorsal nucleus of the lateral lemniscus, from the dorsal nucleus of the lateral lemniscus to the contralateral inferior colliculus, from the sagulum to the contralateral sagulum) reach their targets via the commissure of Probst. The remaining 5 decussations (from the inferior colliculus to the contralateral inferior colliculus or medial geniculate, from the intermediate nucleus of the lateral lemniscus to the contralateral medial geniculate, from the sagulum to the contralateral inferior colliculus or medial geniculate) reach their targets via the commissure of the inferior colliculus. The results also suggest that the commissure of Probst is not a general avenue for decussating auditory fibers of the lateral lemniscus but is instead a specific avenue only for fibers from the dorsal nucleus of the lateral lemniscus and sagulum. The results also show that, in the cat at least, the dorsal nucleus of the lateral lemniscus does not project beyond the inferior colliculus to either the superior colliculus or medial geniculate--the cells previously reported as doing so are probably those of the immediate neighbors of the dorsal nucleus, the intermediate nucleus of the lateral lemniscus and sagulum.

Acoustic chiasm. III: Nature, distribution, and sources of afferents to the lateral superior olive in the cat.

The outcomes of seven experiments are reported, each directed to the nature and sources of the excitation and inhibition impinging on the lateral superior olive (LSO) in cats. In the first experiment, we used conventional 14C 2-DG methods to determine the specificity, precision, and extent of symmetry in the stimulation reaching LSO from the ipsilateral and contralateral ears. In Experiment 2, we sought the presence of GABA and glycine receptors in LSO using conventional, in vitro receptor-binding methods. On the basis of these results, we used in vitro high-affinity uptake methods in Experiment 3 to seek evidence that some of the terminals as well as the receptors in LSO are glycinergic. In Experiment 4, we used immunocytochemical methods to show that the somata known to supply the contralateral projections to LSO, and their terminals in LSO, are each immunoreactive with an antibody directed to a glycine-protein conjugate. In Experiment 5, we made use of a glycinergic neuron's avidity for transporting glycine retrogradely to label the likely sources of the glycinergic terminals in LSO. In Experiment 6, we used immunocytochemical methods to show that the spherical and globular cells of the ventral cochlear nucleus and terminals in LSO and in MTB are glutamatergic and/or aspartergic. In Experiment 7, we used receptor binding methods to determine whether the glutamate/aspartate receptors in LSO are probably of the kainate or of the quisqualate type. The results of the several experiments suggest that probably glutamate-quisqualate synapses mediate LSO's ipsilaterally driven excitatory responses and glycinergic synapses mediate its contralaterally driven inhibitory responses. The two types of input appear to be well matched in LSO's medial and middle limbs with glycinergic terminals mostly perisomatic and glutamatergic terminals mostly peridendritic. However, LSO's low frequency lateral limb appears to be somewhat different; it receives less stimulation from the contralateral ear. Instead, LSO's lateral limb may receive some of its glycinergic input directly from the ipsilateral ventral cochlear nucleus and/or indirectly via the juxtaposed lateral nucleus of the trapezoid body.

Auditory cortex of the long-eared hedgehog (Hemiechinus auritus). I. Boundaries and frequency representation.

The boundaries of the primary auditory cortex of the long-eared hedgehog, Hemiechinus auritus, were determined by single-cell recordings, myeloarchitecture and retrograde horseradish peroxidase labeling in the medial geniculate, using anesthetized animals. The auditory cortex is located on the lateral surface of the temporal cortex, medial to the rhinal fissure. Responses to pure tones revealed an orderly representation of best frequencies in the primary auditory cortex, with low frequencies represented rostrally and high frequencies caudally. A second auditory field caudal to the primary one was indicated.

Neuroanatomical distribution of receptors for three potential inhibitory neurotransmitters in the brainstem auditory nuclei of the cat.

In order to visualize the relative abundance of each of three potentially inhibitory neurotransmitters in the nuclei of the brainstem auditory pathway, receptor sites for glycine, GABA-A, and muscarinic acetylcholine (ACh) have been localized in the cat's brainstem auditory system. Conventional autoradiographic receptor-binding procedures were used and the distributions of the receptors were inferred from the respective distributions of tritiated strychnine, muscimol, and quinuclidinyl benzilate (QNB) binding sites. The results show that glycine may be the major inhibitory neurotransmitter in the auditory system as it ascends to the midbrain in that relatively high levels of strychnine binding are present in every major nucleus of the system. In contrast, high levels of muscimol binding of high-affinity GABA-A receptors are confined mostly to the dorsal cochlear nucleus, the dorsal nucleus of the lateral lemniscus, and the central and cortical regions of the inferior colliculus, while high levels of QNB binding of muscarinic ACh receptors are seen only in the central and cortical regions of the inferior colliculus.

Origin of mammalian thalamocortical projections. I. Telencephalic projections of the medial geniculate body in the opossum (Didelphis virginiana).

Telencephalic projections from the medial geniculate nucleus (MG) in opossum were traced with tritiated leucine autoradiography and by horseradish peroxidase and fluorescent dye retrograde labeling techniques. The results show that the opossum's MG contains two separate populations of neurons-one in the anterior two-thirds of MG projecting to auditory neocortex, the other occupying the entire caudal one-third of MG and projecting mostly to lateral amygdala and putamen. Because the subcortical projection of the MG in opossum is larger than that seen in any other mammal to date, it is reminiscent of the subcortical projections of the MG in reptiles and birds. Furthermore, when the subcortical projections of the MG in reptiles and opossums are compared with similar subcortical projections of the MG in rats, cats, and monkeys, the proportion of the MG neurons projecting to subcortical structures is seen to be inversely related to the recency of each animal's common ancestry with primates. The possibility that the subcortical projection of the MG in mammals is homologous with that seen in reptiles or birds implies that it might be a dwindling vestige of the projection present in the common ancestry of reptiles and mammals.

Origin of interhemispheric fibers in acallosal opossum (with a comparison to callosal origins in rat).

The neocortical origins of the anterior commissure in the acallosal, marsupial opossum were studied with the horseradish peroxidase (HRP) method. Following complete surgical transection of the anterior commissure, HRP was applied directly to the cut fiber tips. This procedure resulted in very large numbers of vividly labeled cells within the neocortex. The labeled cells were plotted and counted for comparison among cytoarchitectonic areas and among cortical layers. For comparative purposes, the neocortical origins of the corpus callosum are studied with the same procedure in the rat. No cytoarchitectonic area was entirely devoid of labeled cells in either species. The concentration of labeled cells throughout the entire neocortex averaged 25.2 cells/0.05 mm3 in opossum and 31.2 cells/0.05 mm3 in rat. The concentrations of labeled cells were correlated for the eight cytoarchitectonic areas common to the two species, though they were different enough in number to be statistically reliable. The distribution of labeled cells both among and within cytoarchitectonic areas was often more homogeneous in opossum than in rat. Although cortical layer 1 had no labeled cells in either species, the distribution of labeled cells across the remaining cortical layers differed sharply between the two species. In opossum, layer 3 had the most labeled cells (averaging 55% of the total number) while layer 5 had considerably less (averaging 12%). In rat, layer 5 had as many labeled cells as layer 3--both layers averaging 43% of the total number of labeled cells. In both species, striate cortex deviated markedly from other cytoarchitectonic areas. Although both species had very few labeled cells in striate cortex, those that were labeled were invariably supragranular in opossum and infragranular in rat. The similarities and dissimilarities in the topographic distribution of the origins of the two types of interhemispheric fiber systems seem to parallel the degree of cortical (and thalamic) differentiation in the two animals. However, the differences in laminar distribution are much greater and in particular, the small contribution of layer 5 in opossum as opposed to rat may well be functionally significant.

Acoustic chiasm II: Anatomical basis of binaurality in lateral superior olive of cat.

The afferent projections to the lateral superior olive (LSO) were examined with horseradish peroxidase, horseradish peroxidase-wheat germ agglutinin conjugate, 125I-wheat germ agglutinin and tritiated leucine autoradiograhy, anterograde axonal degeneration, and 14C-2-deoxyglucose methods. The pathway to the ipsilateral LSO orginates in the spherical cells in anteroventral cochlear nucleus. Although some of the fibers pass above the lateral nucleus of the trapezoid body, most pass below it and turn at right angles to enter the LSO either directly through its ventral, lateral, or dorsal borders, or through its ventral or dorsal hilus. They end in unpolarized terminal fields throughout the LSO. Most if not all of these fibers are true collaterals of axons continuing across the midline in the trapezoid body. Verifying Held's (1893) finding of a major direct projection from the cochlear nucleus to the contralateral medial nucleus of the trapezoid body (MTB) and Rasmussen's ('46) finding of a major projection from the MTB to the LSO, the present results illustrate that this two-neuron pathway probably supplies all but a very small component of the relatively direct input to the LSO from the contralateral ear. This pathway originates in the globular cells of the ventral cochlear nucleus and relays mostly though not exclusively through the "principal cells" in the more rostral parts of the MTB. It terminates mostly in perisomal endings in unpolarized fields throughout the LSO, though most heavily within the (high frequency) medial and middle limbs and less heavily in the LSO's (low frequency) lateral limb. In addition to this indirect pathway, there is a small direct pathway to the contralateral LSO as suggested by Goldberg and Brown ('69) and Warr ('72, '82). This direct pathway to the contralateral LSO, like the direct ipsilateral pathway, probably originates in the spherical cell region of the ventral cochlear nucleus, crosses the midline in the trapezoid body, and terminates in a small circumscribed area within the LSO's ventromedial (high frequency) area. The 2-deoxyglucose method applied to cats in which the ipsilateral and contralateral pathways have been surgically isolated shows that each of the pathways converging on the LSO is topographically and tonotopically organized with the ipsilateral and the combined contralateral terminations in strict tonotopic register.

Acoustic chiasm: efferent projections of the lateral superior olive.

The efferent connections of the cat's lateral superior olive (LSO) were examined first with kainic acid-induced anterograde degeneration and tritiated leucine autoradiography and then by systematic repetition of HRP and fluorescent dye retrograde tract-tracing techniques. The results show that virtually all LSO cells have axons ascending either contralaterally or ipsilaterally to high pontine and midbrain levels of the brainstem. Most terminate in the ventrolateral division of either the ipsilateral or contralateral central nucleus of the inferior colliculus, some terminate in the ipsilateral or contralateral dorsal nucleus of the lateral lemniscus, and a small number terminate in the ipsilateral intermediate nucleus of the lateral lemniscus. Only a small proportion (less than 5%) of LSO cells project to both sides via axon collaterals. The ipsilateral, contralateral, and bilateral projections arise from three overlapping subpopulations of cells within LSO: Those projecting ipsilaterally are concentrated in its lateral limb; those projecting contralaterally are concentrated in its medial limb; the few projecting bilaterally are thinly scattered throughout. Therefore, a lateral-medial gradient is present across LSO based on the laterality of its cell's efferent targets. This gradient parallels LSO's tonotopic gradient: The higher the characteristic frequency of an LSO cell, the more likely it is to project contralaterally. This arrangement of LSO's ascending projections, with most of its lateral cells projecting ipsilaterally and most of its medial cells projecting contralaterally, is similar to the arrangement of the optic chiasm in animals with overlapping eye-fields. Its presence seems to provide an anatomical basis for some recent electrophysiological and behavioral reports of chiasm-like properties of the superior olivary complex.

Ascending auditory afferents to the nuclei of the lateral lemniscus.

Afferents from the hindbrain auditory system to the nuclei of the lateral lemniscus were analyzed by the use of orthograde and retrograde axon-tracing techniques. Three divisions of the nuclei of the lateral lemniscus, a dorsal, an intermediate, and a ventral division are discussed. The dorsal nucleus of the lateral lemniscus is a recipient of afferents from cells located mainly in the superior olivary complex and the contralateral dorsal nucleus of the lateral lemniscus. It receives direct afferents from only a few cells in the cochlear nuclei. In sharp contrast, the ventral nucleus of the lateral lemniscus is the recipient of afferents from many cells in the contralateral ventral cochlear nucleus and from only a few cells in the superior olivary complex. Further, it receives no afferents from cells in the contralateral nuclei of the lateral lemniscus. The intermediate nucleus of the lateral lemniscus receives afferents from some cells in the cochlear nucleus and the superior olivary complex. It is unique among the three nuclei of the lateral lemniscus in that it receives a substantial projection from the medial nucleus of the trapezoid body.

Accessory abducens nucleus and its relationship to the accessory facial and posterior trigeminal nuclei in cat.

Retrograde transport of HRP by the abducens nerve results in the labelling of its principal nucleus and, in addition, a second nucleus about 2.5 mm ventrolateral to the principal nucleus. The presence of this second or accessory nucleus of the abducens in a mammal confirms the observations of several Nineteenth Century anatomists and rebuts the conclusions of more recent investigators who argued that the nucleus was allied instead to the facial or trigeminal nerves. The same HRP technique applied to the facial or trigeminal nerves shows that the accessory nucleus of the abducens is in the same parasagittal plane as the accessory nucleus of the facial nerve and the most caudal cells of the motor trigeminal nucleus. The accessory abducens and accessory facial nuclei fall in a ventrocaudal to dorsorostral line between the principal nucleus of the facial and the motor nucleus of the trigeminal with the accessory abducens just caudal and ventral to the accessory facial.