Inhibitory Projections from the Inferior Colliculus to the Medial Geniculate body Originate from Four Subtypes of GABAergic Cells.

GABAergic cells constitute 20-40% of the cells that project from the inferior colliculus [(IC) a midbrain auditory hub] to the medial geniculate body [(MG) the main auditory nucleus of the thalamus]. Four subtypes of GABAergic IC cells have been identified based on their association with perineuronal nets (PNs) and dense rings of axosomatic terminals expressing vesicular glutamate transporter 2 (VGLUT2 rings). These subtypes differ in their soma size and distribution within the IC. Based on previous work emphasizing large GABAergic cells as the origin of GABAergic IC-MG projections, we hypothesized that GABAergic IC cells surrounded by PNs and VGLUT2 rings, which tend to have larger somas, were more likely to project to the MG than smaller cells lacking these extracellular markers. Here, we injected retrograde tract tracers into the MG of guinea pigs of either sex and analyzed retrogradely labeled GABAergic cells in the ipsilateral IC for soma size and association with PNs and/or VGLUT2 rings. We found a range of GABAergic soma sizes present within the IC-MG pathway, which were reflective of the full range of GABAergic soma sizes present within the IC. Further, we found that all four subtypes of GABAergic IC cells participate in the IC-MG pathway, and that GABAergic cells lacking PNs and VGLUT2 rings were more prevalent within the pathway than would be expected based on their overall prevalence in the IC. These results may provide an anatomical substrate for the multiple roles of inhibition in the IC-MG pathway, which have emerged in electrophysiological studies.

Analysis of excitatory synapses in the guinea pig inferior colliculus: a study using electron microscopy and GABA immunocytochemistry.

The inferior colliculus (IC) integrates ascending auditory input from the lower brainstem and descending input from the auditory cortex. Understanding how IC cells integrate these inputs requires identification of their synaptic arrangements. We describe excitatory synapses in the dorsal cortex, central nucleus, and lateral cortex of the IC (ICd, ICc and IClc) in guinea pigs. We used electron microscopy (EM) and post-embedding anti-GABA immunogold histochemistry on aldehyde-fixed tissue from pigmented adult guinea pigs. Excitatory synapses were identified by round vesicles, asymmetric synaptic junctions, and gamma-aminobutyric acid-immunonegative (GABA-negative) presynaptic boutons. Excitatory synapses constitute ∼60% of the synapses in each IC subdivision. Three types can be distinguished by presynaptic profile area and number of mitochondrial profiles. Large excitatory (LE) boutons are more than 2 µm(2) in area and usually contain five or more mitochondrial profiles. Small excitatory (SE) boutons are usually less than 0.7 µm(2) in area and usually contain 0 or 1 mitochondria. Medium excitatory (ME) boutons are intermediate in size and usually contain 2 to 4 mitochondria. LE boutons are mostly confined to the ICc, while the other two types are present throughout the IC. Dendritic spines are the most common target of excitatory boutons in the IC dorsal cortex, whereas dendritic shafts are the most common target in other IC subdivisions. Finally, each bouton type terminates on both gamma-aminobutyric acid-immunopositive (GABA+) and GABA-negative (i.e., glutamatergic) targets, with terminations on GABA-negative profiles being much more frequent. The ultrastructural differences between the three types of boutons presumably reflect different origins and may indicate differences in postsynaptic effect. Despite such differences in origins, each of the bouton types contact both GABAergic and non-GABAergic IC cells, and could be expected to activate both excitatory and inhibitory IC circuits.

Cholinergic cells in the tegmentum send branching projections to the inferior colliculus and the medial geniculate body.

The pontomesencephalic tegmentum (PMT) provides cholinergic input to the inferior colliculus (IC) and the medial geniculate body (MG). PMT cells are often characterized as projecting to more than one target. The purpose of this study was to determine whether individual PMT cholinergic cells, (1) innervate the auditory pathways bilaterally via collateral projections to left and right auditory thalamus; or, (2) innervate multiple levels of the auditory pathways via collateral projections to the auditory thalamus and inferior colliculus. We used multiple retrograde tracers to identify individual PMT cells that project to more than one target. We combined the retrograde tracer studies with immunohistochemistry for choline acetyltransferase to determine whether the projecting cells were cholinergic. We found that individual PMT cells send branching axonal projections to two or more auditory targets in the midbrain and thalamus. The collateral projection pattern that we observed most frequently was to the ipsilateral IC and ipsilateral MG. Cells projecting to both MGs were somewhat less common, followed by cells projecting to the contralateral IC and ipsilateral MG. Both cholinergic and non-cholinergic cells contribute to each of these projection patterns. Less often, we found cells that project to one IC and both MGs; there was no evidence for non-cholinergic cells in this projection pattern. It is likely that collateral projections from PMT cells could have coordinated effects bilaterally and at multiple levels of the ascending auditory pathways.

Multiple origins of cholinergic innervation of the cochlear nucleus.

Acetylcholine (Ach) affects a variety of cell types in the cochlear nucleus (CN) and is likely to play a role in numerous functions. Previous work in rats suggested that the acetylcholine arises from cells in the superior olivary complex, including cells that have axonal branches that innervate both the CN and the cochlea (i.e. olivocochlear cells) as well as cells that innervate only the CN. We combined retrograde tracing with immunohistochemistry for choline acetyltransferase to identify the source of ACh in the CN of guinea pigs. The results confirm a projection from cholinergic cells in the superior olivary complex to the CN. In addition, we identified a substantial number of cholinergic cells in the pedunculopontine tegmental nucleus (PPT) and the laterodorsal tegmental nucleus (LDT) that project to the CN. On average, the PPT and LDT together contained about 26% of the cholinergic cells that project to CN, whereas the superior olivary complex contained about 74%. A small number of additional cholinergic cells were located in other areas, including the parabrachial nuclei.The results highlight a substantial cholinergic projection from the pontomesencephalic tegmentum (PPT and LDT) in addition to a larger projection from the superior olivary complex. These different sources of cholinergic projections to the CN are likely to serve different functions. Projections from the superior olivary complex are likely to serve a feedback role, and may be closely tied to olivocochlear functions. Projections from the pontomesencephalic tegmentum may play a role in such things as arousal and sensory gating. Projections from each of these areas, and perhaps even the smaller sources of cholinergic inputs, may be important in conditions such as tinnitus as well as in normal acoustic processing.

Substrates of auditory frequency integration in a nucleus of the lateral lemniscus.

In the intermediate nucleus of the lateral lemniscus (INLL), some neurons display a form of spectral integration in which excitatory responses to sounds at their best frequency are inhibited by sounds within a frequency band at least one octave lower. Previous work showed that this response property depends on low-frequency-tuned glycinergic input. To identify all sources of inputs to these INLL neurons, and in particular the low-frequency glycinergic input, we combined retrograde tracing with immunohistochemistry for the neurotransmitter glycine. We deposited a retrograde tracer at recording sites displaying either high best frequencies (>75 kHz) in conjunction with combination-sensitive inhibition, or at sites displaying low best frequencies (23-30 kHz). Most retrogradely labeled cells were located in the ipsilateral medial nucleus of the trapezoid body (MNTB) and contralateral anteroventral cochlear nucleus. Consistent labeling, but in fewer numbers, was observed in the ipsilateral lateral nucleus of the trapezoid body (LNTB), contralateral posteroventral cochlear nucleus, and a few other brainstem nuclei. When tracer deposits were combined with glycine immunohistochemistry, most double-labeled cells were observed in the ipsilateral MNTB (84%), with fewer in LNTB (13%). After tracer deposits at combination-sensitive recording sites, a striking result was that MNTB labeling occurred in both medial and lateral regions. This labeling appeared to overlap the MNTB labeling that resulted from tracer deposits in low-frequency recording sites of INLL. These findings suggest that MNTB is the most likely source of low-frequency glycinergic input to INLL neurons with high best frequencies and combination-sensitive inhibition. This work establishes an anatomical basis for frequency integration in the auditory brainstem.

Projections from auditory cortex to midbrain cholinergic neurons that project to the inferior colliculus.

We have shown that auditory cortex projects to cholinergic cells in the pedunculopontine tegmental nucleus (PPT) and laterodorsal tegmental nucleus (LDT). PPT and LDT are the sources of cholinergic projections to the inferior colliculus, but it is not known if the cortical inputs contact the cholinergic cells that project to the inferior colliculus. We injected FluoroRuby into auditory cortex in pigmented guinea pigs to label cortical projections to PPT and LDT. In the same animals, we injected Fast Blue into the left or right inferior colliculus to label PPT and LDT cells that project to the inferior colliculus. We processed the brain to identify cholinergic cells with an antibody to choline acetyltransferase, which was visualized with a green fluorescent marker distinguishable from both FluoroRuby and Fast Blue. We then examined the PPT and LDT to determine whether boutons of FluoroRuby-labeled cortical axons were in close contact with cells that were double-labeled with the retrograde tracer and the immunolabel. Apparent contacts were observed ipsilateral and, less often, contralateral to the injected cortex. On both sides, the contacts were more numerous in PPT than in LDT. The results indicate that auditory cortex projects directly to brainstem cholinergic cells that innervate the ipsilateral or contralateral inferior colliculus. This suggests that cortical projections could elicit cholinergic effects on both sides of the auditory midbrain.

Sources of cholinergic input to the inferior colliculus.

We combined retrograde tracing with immunohistochemistry for choline acetyltransferase to identify the source of cholinergic input to the inferior colliculus (IC) in guinea pigs. Injection of a retrograde tracer into one IC labeled cells in many brainstem nuclei. Retrogradely-labeled cells that were also immunoreactive for choline acetyltransferase were identified in two nuclei in the midbrain tegmentum: the pedunculopontine tegmental nucleus (PPT) and the laterodorsal tegmental nucleus (LDT). More PPT and LDT cells project ipsilaterally than contralaterally to the IC and, on both sides, there are more projecting cells in the PPT than in the LDT. Double-labeled cells were not found in any other brainstem nucleus. A common feature of cholinergic cells in PPT and LDT is collateral projections to multiple targets. We placed different retrograde tracers into each IC to identify cells in PPT and LDT that project to both ICs. In both PPT and LDT, a substantial proportion (up to 57%) of the immunoreactive cells that contained tracer from the contralateral IC also contained tracer from the ipsilateral IC. We conclude that acetylcholine in the IC originates from the midbrain tegmental cholinergic nuclei: PPT and LDT. These nuclei are known to participate in arousal, the sleep/wake cycle and prepulse inhibition of acoustic startle. It is likely that the cholinergic input to the IC is directly associated with these functions.

Projections to the inferior colliculus from layer VI cells of auditory cortex.

A large injection of a retrograde tracer into the inferior colliculus of guinea pigs labeled two bands of cells in the ipsilateral auditory cortex: a dense band of cells in layer V and a second band of cells in layer VI. On the contralateral side, labeled cells were restricted to layer V. The ipsilateral layer VI cells were distributed throughout temporal cortex, suggesting projections from multiple auditory areas. The layer VI cells included pyramidal cells as well as several varieties of non-pyramidal cells. Small tracer injections restricted to the dorsal cortex or external cortex of the inferior colliculus consistently labeled cells in layer VI. Injections restricted to the central nucleus of the inferior colliculus labeled layer VI cells only rarely. Overall, 10% of the cells in temporal cortex that project to the ipsilateral inferior colliculus were located in layer VI, suggesting that layer VI cells make a significant contribution to the corticocollicular pathway.

Distribution of cholinergic cells in guinea pig brainstem.

We used an antibody to choline acetyltransferase (ChAT) to label cholinergic cells in guinea pig brainstem. ChAT-immunoreactive (IR) cells comprise several prominent groups, including the pedunculopontine tegmental nucleus, laterodorsal tegmental nucleus, and parabigeminal nucleus, as well as the cranial nerve somatic motor and parasympathetic nuclei. Additional concentrations are present in the parabrachial nuclei and superior colliculus. Among auditory nuclei, the majority of ChAT-IR cells are in the superior olive, particularly in and around the lateral superior olive, the ventral nucleus of the trapezoid body and the superior paraolivary nucleus. A discrete group of ChAT-IR cells is located in the sagulum, and additional cells are scattered in the nucleus of the brachium of the inferior colliculus. A group of ChAT-IR cells lies dorsal to the dorsal nucleus of the lateral lemniscus. A few ChAT-IR cells are found in the cochlear nucleus and the ventral nucleus of the lateral lemniscus. The distribution of cholinergic cells in guinea pigs is largely similar to that of other species; differences occur mainly in cell groups that have few ChAT-IR cells. The results provide a basis for further studies to characterize the connections of these cholinergic groups.

On the use of retrograde tracers for identification of axon collaterals with multiple fluorescent retrograde tracers.

A common method for identifying collateral projections is to inject different retrograde tracers into two targets and examine labeled cells for the presence of both tracers. Double-labeled cells are considered to have collateral projections to the two injection sites. This method is widely considered to underestimate the extent of collaterals. To test the efficiency of double-labeling, we mixed equal volumes of two tracers, injected them into one site in a guinea-pig brain, and counted the resulting labeled cells. Ideally, the tracers would have precisely overlapping injection sites and all labeled cells would contain both tracers. We tested several combinations of tracers: 1) Fast Blue and fluorescein dextran; 2) fluorescein dextran and FluoroGold; 3) fluorescein dextran and FluoroRuby; 4) FluoroGold and green beads; 5) FluoroGold and red beads; 6) FluoroRuby and green beads; and, 7) green beads and red beads. For each combination, a mixture was injected into the left inferior colliculus. After 1 week to allow for transport, labeled cells were counted in the right inferior colliculus and the left temporal cortex. For each mixture, the results were similar for the two areas. The percentage of cells that were double-labeled varied from 0% to 100%, depending on tracer combination. The highest efficiencies (>96%) were observed with red beads and green beads or with FluoroRuby and fluorescein dextran. The limited efficiency of other mixtures could be accounted for only in part by incomplete overlap of the two tracers at the injection site. The results indicate that the specific combination of tracers used to search for collateral projections can greatly affect the findings.

Origins of projections from the inferior colliculus to the cochlear nucleus in guinea pigs.

We used retrograde tracing techniques to examine the projections from the inferior colliculus to the cochlear nucleus in guinea pigs. Following injection of a retrograde tracer into one cochlear nucleus, labeled cells were found bilaterally in all subdivisions of the inferior colliculus. The majority of cells were located in the central nucleus and external cortex; relatively few cells were located in the dorsal cortex. Multipolar (stellate) cells were labeled in all subdivisions of the inferior colliculus. In the central nucleus, disk-shaped cells were also labeled. To determine whether individual collicular neurons send collateral projections to the cochlear nuclei on both sides, we injected different fluorescent tracers into left and right cochlear nuclei in the same animal. The inferior colliculi contained very few double-labeled cells, indicating that the projections to ipsilateral and contralateral cochlear nuclei originate from separate populations of cells.

Afferent projections of the superior olivary complex.

Based on current literature, the afferents of the superior olivary complex (SOC) are described including those from the cochlear nucleus, inferior colliculus, thalamus, and auditory cortex. Intrinsic SOC afferents and non-auditory afferents from the serotoninergic and noradrenergic systems are also described. New data are provided that show a differential distribution of serotoninergic afferents within the SOC: serotoninergic fibers were relatively sparse in the lateral and medial superior olives and the medial nucleus of the trapezoid body and were most numerous in periolivary regions. There are variations in the density of serotoninergic fibers within periolivary regions themselves. New data is also provided on auditory and non-auditory afferents to SOC neurons, which have known targets. These include: cochlear nucleus afferents to periolivary (lateral nucleus of the trapezoid body, LNTB) cells that project to the inferior colliculus; cortical afferents to periolivary (ventral nucleus of the trapezoid body, VNTB) cells that project to the cochlear nucleus; and serotoninergic and noradrenergic afferents to periolivary (LNTB and VNTB) cells that project to the cochlear nucleus. The relationships between other types of afferents and SOC neurons with known projections are also described as functional circuits. The circuits include those that are part of the ascending auditory system (to the inferior and superior colliculi, lateral lemniscus, and medial geniculate nucleus), the descending auditory system (to the cochlea and cochlear nucleus), and the middle ear reflex circuits.

Descending auditory pathways: projections from the inferior colliculus contact superior olivary cells that project bilaterally to the cochlear nuclei.

Multiple retrograde and anterograde tracers were used to characterize a pathway that extends from the inferior colliculus to both the left and right cochlear nuclei via a synaptic relay in the superior olivary complex. Different fluorescent tracers were injected into the left and right cochlear nuclei to identify cells in the superior olivary complex that project bilaterally. Double-labeled cells were present in almost all periolivary nuclei; the majority were located in the ventral nucleus of the trapezoid body and the anteroventral periolivary nucleus. Because these two nuclei are targets of descending projections from the inferior colliculus, triple-labeling experiments were performed to determine whether collicular axons contact the periolivary cells that project to the cochlear nuclei. The results demonstrate that descending axons from the inferior colliculus contact periolivary cells that project to the cochlear nuclei, including periolivary cells that project bilaterally. This pathway could provide an opportunity for higher levels of the auditory system to influence activity bilaterally in the cochlear nuclei and thus to modulate the initial processing of acoustic information by the brain.

Ventral nucleus of the lateral lemniscus in guinea pigs: cytoarchitecture and inputs from the cochlear nucleus.

Cytoarchitectonic criteria were used to distinguish three subdivisions of the ventral nucleus of the lateral lemniscus in guinea pigs. Axonal tracing techniques were used to examine the projections from the cochlear nucleus to each subdivision. Based on the cell types they contain and their patterns of input, we distinguished ventral, dorsal, and anterior subdivisions of the ventral nucleus of the lateral lemniscus. All three subdivisions receive bilateral inputs from the cochlear nucleus, with contralateral inputs greatly outnumbering ipsilateral inputs. However, the relative density of the inputs varies: the ventral subdivision receives the densest projection, whereas the anterior subdivision receives the sparsest projection. Further differences are apparent in the morphology of the afferent axons. Following an injection of Phaseolus vulgaris-leucoagglutinin into the ventral cochlear nucleus, most of the axons on the contralateral side and all of the axons on the ipsilateral side are thin. Thick axons are present only in the ventral subdivision contralateral to the injection site. The evidence from both anterograde and retrograde tracing studies suggests that the thick axons originate from octopus cells, whereas the thin axons arise from multipolar cells and spherical bushy cells. The differences in constituent cell types and in patterns of inputs suggest that each of the three subdivisions of the ventral nucleus of the lateral lemniscus makes a distinct contribution to the analysis of acoustic signals.

Projections from the ventral cochlear nucleus to the inferior colliculus and the contralateral cochlear nucleus in guinea pigs.

Multipolar cells in the ventral cochlear nucleus are the source of projections to numerous brainstem auditory nuclei, including the contralateral and ipsilateral inferior colliculi and the contralateral cochlear nucleus. Multiple fluorescent tracers were used to label the multipolar cells that project to each of these targets. Following injections of different tracers into each target, the ventral cochlear nucleus was examined for the presence of cells that contained more than one tracer. Such cells were never observed. In contrast, double-labeled cells were common in the dorsal cochlear nucleus, where cells frequently contained the two tracers that were injected into the ipsilateral and contralateral inferior colliculi. The distribution and somatic morphology of cells in the ventral cochlear nucleus that project to each of the three targets were examined. Each population contained cells with somas that ranged in shape from elongated to rounded, but there were differences in soma size. Projections to the ipsilateral and contralateral inferior colliculi arise predominantly from small to medium-sized cells, the average size being slightly less for cells with projections to the ipsilateral colliculus. Projections to the contralateral cochlear nucleus arise from cells with somas that range in size from small to large, including cells much larger than those that projected to either inferior colliculus. On the basis of these results, we conclude that projections from the ventral cochlear nucleus to the ipsilateral and contralateral inferior colliculi and to the contralateral cochlear nucleus arise in three different populations of multipolar cells.

Origins and targets of commissural connections between the cochlear nuclei in guinea pigs.

Our objective was to identify the origins and targets of axons that project from one cochlear nucleus to the other. First, retrograde tracers were injected into one cochlear nucleus to label commissural cells in the opposite nucleus. In the dorsal cochlear nucleus, a few cells in the deep layers were labeled; they were not further classified according to type. In the ventral cochlear nucleus, all commissural cells that could be classified were multipolar cells. Second, an anterograde tracer was injected into one cochlear nucleus, and the distribution of boutons in the opposite cochlear nucleus was examined. Labeled boutons were present throughout the ventral cochlear nucleus, where they appeared to contact multipolar cells, spherical and globular bushy cells, and octopus cells. In the dorsal cochlear nucleus, labeled boutons were present in the fusiform cell and deep layers and appeared to contact fusiform cells and cells of unknown type. Many labeled terminals were also present in the granule cell regions. Injections into regions associated with high or low frequencies labeled boutons in corresponding regions in the contralateral ventral cochlear nucleus. Third, multiple tracers were used to determine whether cells that project to the inferior colliculus are contacted by commissural axons. Boutons labeled by anterograde transport of one tracer placed in the cochlear nucleus were frequently observed to be apposed to cells that were labeled by retrograde transport of a different tracer placed in the contralateral inferior colliculus. We conclude that commissural projections originate from multipolar cells throughout the ventral cochlear nucleus (and from a small number of cells in the dorsal cochlear nucleus) and make contact with all major cell types of the cochlear nuclei, including at least some of those that project to the inferior colliculus.

Projections from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs.

Axonal tracing techniques were used to study the projection from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs. Different tracers were used to identify the cell types that give rise to the projections, the morphology of their axons, and the cell types that they contact in the superior paraolivary nucleus. Injections of Fluoro-Gold or peroxidase-labeled-WGA and HRP into the superior paraolivary nucleus labeled multipolar cells and octopus cells bilaterally in the ventral cochlear nucleus, mainly on the contralateral side. Injections of PHAL into the ventral cochlear nucleus labeled two types of axons in the superior paraolivary nucleus. Thin axons branch infrequently and give rise primarily to small, en passant boutons. Thick axons have larger boutons, many of which are terminal boutons that arise from short collaterals. Thin axons appear to originate from multipolar cells, whereas thick axons probably originate from octopus cells. Both types are found bilaterally after an injection into the ventral cochlear nucleus on one side. Individual thick or thin axons may contact multiple cell types in the superior paraolivary nucleus. Individual cells in the superior paraolivary nucleus can receive convergent input from both thick and thin axons. Combined anterograde and retrograde transport of different fluorescent tracers was used to identify the projections of the cells in the superior paraolivary nucleus that receive inputs from the ventral cochlear nucleus. Cells in the superior paraolivary nucleus that projected to the ipsilateral cochlear nucleus or to the ipsilateral inferior colliculus appeared to be contacted by axons that were labeled by anterograde transport from the contralateral ventral cochlear nucleus. Thus the projections to the superior paraolivary nucleus are in a position to affect the activity in both ascending and descending auditory pathways.

Projections to the cochlear nuclei from principal cells in the medial nucleus of the trapezoid body in guinea pigs.

Spherical and globular cells in the cochlear nucleus provide input to the cell groups in the superior olivary complex devoted to the analysis of binaural cues. Descending projections from the superior olivary complex appear to inhibit the spherical and globular cells. It is not known which of the numerous cell types in the superior olive provide this descending input, but recent studies have shown that some of the cells are located in the medial nucleus of the trapezoid body (MTB). The present experiments were designed to determine whether the MTB projections arise from principal cells, which are known to play a role in sound localization, and to determine whether their projections terminate on spherical or globular cells. Principal cells in the MTB are characterized by their contacts with synaptic specializations called calyces, which arise from the axons of cells in the contralateral cochlear nucleus. In the first experiment, a fluorescent tracer was injected into one cochlear nucleus to label the calyces anterogradely. A different tracer was injected into the opposite cochlear nucleus to label cells retrogradely in the MTB. In every case, some of the labeled cells were enveloped by a labeled calyx, demonstrating that principal cells do project to the cochlear nucleus. In the second experiment, fluorescent tracers were injected into different parts of the cochlear nucleus. Analysis of the distribution of labeled cells suggested that MTB projections selectively target the globular cell region of the cochlear nucleus. In a third experiment, the axonal arborizations arising from this projection were labeled with biocytin or wheat germ agglutinin conjugated to horseradish peroxidase. Labeled boutons appeared to contact globular cells but not spherical cells. Multipolar cells in the ventral cochlear nucleus and cells in the dorsal cochlear nucleus were also contacted. The results suggest that MTB projections to the cochlear nucleus arise largely from principal cells and contact, at least in part, cells in the cochlear nucleus that give rise to ascending pathways involved in sound localization.

The sublaminar organization of corticogeniculate neurons in layer 6 of macaque striate cortex.

We examined the laminar distribution of corticogeniculate neurons in the macaque striate cortex labeled by axonal transport following injections of retrograde tracers into the lateral geniculate nucleus (LGN). Large injections of retrograde tracers involving all layers of the LGN resulted in a distinctive bilaminar distribution of labeled cells in cortical layer 6. One tier of labeled neurons was located along the layer 5-6 border and a second was located near the bottom of the layer, leaving the middle of layer 6 largely free of labeled neurons. Following injections of tracers that were restricted to the magnocellular layers of the LGN, almost all of the labeled neurons were located in the lower tier. In contrast, following injections of retrograde tracers confined to the parvocellular layers of the LGN, labeled cells were found in both tiers, with the greatest number in the upper tier. Thus, layer 6 of macaque striate cortex consists of three distinct sublayers only two of which are the source of descending projections to the LGN: an upper tier that projects exclusively to the parvocellular layers and a lower tier that projects to both magnocellular and parvocellular layers.

Organization of the superior olivary complex in the guinea pig: II. Patterns of projection from the periolivary nuclei to the inferior colliculus.

The superior olivary complex is a major source of auditory projections to the inferior colliculus. Although the projections from the medial and lateral superior olivary nuclei have been well characterized, projections from the surrounding periolivary nuclei have received relatively little attention. In the guinea pig, cytoarchitectonic criteria can be used to distinguish 11 periolivary nuclei that can be divided into four groups. These are: 1) a lateral group that comprises the anterolateral and posteroventral periolivary nuclei and the lateral nucleus of the trapezoid body; 2) a dorsal group that comprises the dorsal and dorsolateral periolivary nuclei; 3) a ventral group that comprises the rostral, ventromedial, and anteroventral periolivary nuclei and the ventral nucleus of the trapezoid body; and 4) a medial group that comprises the superior paraolivary nucleus and the medial nucleus of the trapezoid body. In the present study we used horseradish peroxidase and fluorescent tracers to identify olivocollicular cells in each of the periolivary nuclei. The lateral, dorsal, and medial periolivary groups project bilaterally, and the ventral periolivary group projects ipsilaterally. Within groups, individual nuclei contain different numbers of olivocollicular cells. The posteroventral periolivary nucleus is the only periolivary nucleus that does not project to the inferior colliculus. The superior paraolivary nucleus is the only periolivary nucleus that contains significant numbers of individual cells that project to both inferior colliculi. The remaining periolivary nuclei project only ipsilaterally or contain separate populations of cells that project to the two inferior colliculi.