Functional networks of parvalbumin-immunoreactive neurons in cat auditory cortex.

Inhibitory interneurons constitute ∼20% of auditory cortical cells and are essential for shaping sensory processing. Connectivity patterns of interneurons in relation to functional organization principles are not well understood. We contrasted the connection patterns of parvalbumin-immunoreactive cells in two functionally distinct cortical regions: the tonotopic, narrowly frequency-tuned module [central narrow band (cNB)] of cat central primary auditory cortex (AI) and the nontonotopic, broadly tuned second auditory field (AII). Interneuronal connectivity patterns and laminar distribution were identified by combining a retrograde tracer (wheat-germ agglutinin apo-horseradish peroxidase colloidal gold) with labeling of the Ca(2+) binding protein parvalbumin (Pv), a marker for the GABAergic interneurons usually described physiologically as fast-spiking neurons. In AI, parvalbumin-positive (Pv+) cells constituted 13% of the retrograde labeled cells in the immediate vicinity of the injection site, compared to 10% in AII. The retrograde labeling of Pv+ cells along isofrequency countours was confined to the cNB. The spatial spread of labeled excitatory neurons in AI was more than twice that found for Pv+ cells. By contrast, in the AII, the spread of Pv+ cells was nearly equal to that of excitatory neurons. The retrograde labeling of Pv+ cells was anisotropic in AI and isotropic in AII. This demonstration of inhibitory networks in auditory cortex reveals that the connections of cat GABAergic AI and AII cells follow different anatomical plans and thus contribute differently to the shaping of neural response properties. The finding that local connectivity of parvalbumin-immunoreactive neurons in AI is closely aligned with spectral integration properties demonstrates the critical role of inhibition in creating distinct processing modules in AI.

Wiring of divergent networks in the central auditory system.

Divergent axonal projections are found throughout the central auditory system. Here, we evaluate these branched projections in terms of their types, distribution, and putative physiological roles. In general, three patterns of axon collateralization are found: intricate local branching, long-distance collaterals, and branched axons (BAs) involved in feedback-control loops. Local collaterals in the auditory cortex may be involved in local processing and modulation of neuronal firing, while long-range collaterals are optimized for wide-dissemination of information. Rarely do axons branch to both ascending and descending targets. Branched projections to two or more widely separated nuclei or areas are numerically sparse but widespread. Finally, branching to contralateral targets is evident at multiple levels of the auditory pathway and may enhance binaural computations for sound localization. These patterns of axonal branching are comparable to those observed in other modalities. We conclude that the operations served by BAs are area- and nucleus-specific and may complement the divergent unbranched projections of local neuronal populations.

Distinct core thalamocortical pathways to central and dorsal primary auditory cortex.

The cat primary auditory cortex (AI) is usually assumed to form one continuous functional region. However, the dorsal and central parts of the AI iso-frequency domain contain neurons that have distinct response properties to acoustic stimuli. In this study, we asked whether neurons projecting to dorsal versus central regions of AI originate in different parts of the medial geniculate body (MGB). Spike rate responses to variations in the sound level and frequency of pure tones were used to measure characteristic frequency (CF) and frequency resolution. These were mapped with high spatial density in order to place retrograde tracers into matching frequency regions of the central narrow-band region (cNB) and dorsal AI. Labeled neurons projecting to these two parts of AI were concentrated in the middle and rostral thirds of the MGB, respectively. There was little evidence that differences in dorsal and central AI function could be due to convergent input from cells outside the ventral division of the MGB (MGBv). Instead, inputs arising from different locations along the caudal-to-rostral dimension of MGBv represent potential sources of response differences between central and dorsal sub-regions of AI.

Local connection patterns of parvalbumin-positive inhibitory interneurons in rat primary auditory cortex.

In the auditory cortex (AC), GABAergic neurons constitute approximately 15-25% of all neurons. GABAergic cells are present in all sensory modalities and essential for modulating sensory receptive fields. Parvalbumin (PV) positive cells represent the largest sub-group of the GABAergic population in auditory neocortex. We investigated the projection pattern of PV cells in rat primary auditory cortex (AI) with a retrograde tracer (wheat germ apo-HRP conjugated to gold [WAHG]) and immunocytochemistry for PV. All AC layers except layer I contained cells double-labeled for PV and WAHG. All co-localized PV+ cells were within 2 mm of the injection site, regardless of laminar origin. Most (ca. 90%) of the co-localized PV cells were within 500 µm of the injection site in both dorsal-ventral and rostral-caudal dimension of the auditory core region. WAHG-only cells declined less rapidly with distance and were found up to 6 mm from the deposit sites. WAHG-only labeled cells in the medial geniculate body were in ventral division loci compatible with an injection in AI. Differences in the range and direction of the distribution pattern of co-localized PV+ cells and WAHG-only cells in AI express distinct functional convergence patterns for the two cell populations.

Convergence of thalamic and cortical pathways in cat auditory cortex.

Cat auditory cortex (AC) receives input from many thalamic nuclei and cortical areas. Previous connectional studies often focused on one connectional system in isolation, limiting perspectives on AC computational processes. Here we review the convergent thalamic, commissural, and corticocortical projections to thirteen AC areas in the cat. Each input differs in strength and may thus serve unique roles. We compared the convergent intrinsic and extrinsic input to each area quantitatively. The intrinsic input was almost half the total. Among extrinsic projections, ipsilateral cortical sources contributed 75%, thalamic input contributed 15%, and contralateral sources contributed 10%. The patterns of distribution support the division of AC areas into families of tonotopic, non-tonotopic, multisensory, and limbic-related areas, each with convergent input arising primarily from within its group. The connections within these areal families suggest a form of processing in which convergence of input to an area could enable new forms of integration. In contrast, the lateral connections between families could subserve integration between categorical representations, allowing otherwise independent streams to communicate and thereby coordinating operations over wide spatial and functional scales. These patterns of serial and interfamilial cooperation challenge more classical models of organization that underestimate the diversity and complexity of AC connectivity.

Microvascular organization of the cat inferior colliculus.

Brain neural activity depends critically on the blood supply to a given structure. The blood supply can differ within and between divisions, which may have functional significance. We analyzed the microvascular organization of the cat inferior colliculus (IC) to determine if the capillary distribution is homogenous throughout. The IC consists of the central nucleus (CN), the dorsal cortex (DC), and the lateral cortex (LC), each with different roles in auditory behavior and perception. Plastic-embedded tissue was studied from adult cats in 1-µm thick semi-thin sections stained with toluidine blue; tissue was sampled from the IC in a caudal-rostral series of sections. The architectonic subdivisions were drawn independently based on Golgi impregnations. We used the nearest neighbor distance (NND) method to quantify capillary density between subdivisions. Overall, the distribution of capillary density was non-homogenous across the IC. We found significant capillary NND differences between the CN and LC (Mann-Whitney test; p ≤ 0.05), CN and DC (Mann-Whitney test; p ≤ 0.05), and LC and DC (Mann-Whitney test; p ≤ 0.05). The CN had the lowest NND values among all three divisions, indicating the highest capillary density. NND values changed gradually as analysis moved from the center of the IC towards the periphery. The significantly higher microvascular density in the CN may imply that the lemniscal auditory pathway has higher levels of blood flow and metabolic activity than non-lemniscal areas of the IC. The non-homogenous microvascular organization of the IC supports parcellation schemes that delineate three major subdivisions and confirms that the borders between the three regions are not sharp.

GABAergic organization of the auditory cortex in the mustached bat (Pteronotus p. parnellii).

The structure and distribution of neurons and axon terminals (puncta) immunostained for gamma-aminobutyric acid (GABA) in the parietotemporal neocortex of the mustached bat (Pteronotus p. parnellii) was studied. The types of GABAergic neurons and puncta (putative terminals) were analyzed, and the immunocytochemical patterns were compared to those in cat auditory cortex (AC). The classic map of mustached bat primary auditory cortex (AI) corresponds to a belt of granular six-layered cortex on the temporal convexity. This area encompasses the Doppler-shifted constant frequency 60 kHz domain (DSCF) described in physiological investigations, as well as its flanking, low-frequency, posterior field (AIp) and the anterior high-frequency region (AIa). Many types of GABAergic neurons correspond to those in cat primary AC. However, the bat had a significantly lower proportion of such cells in five of the six layers. The classes of GABAergic neurons in most layers were small, medium-sized, and large multipolar cells, and bipolar and bitufted neurons. Types found in only one or two layers included horizontal cells (layers I and VI) or extraverted multipolar neurons (layer II). Only layer IV had comparable percentages (∼ 26%), suggesting that the GABAergic influence on lemniscal thalamocortical input is conserved phylogenetically. While the cellular basis for GABAergic cortical processing may reflect shared neural circuits and common modes of inhibitory processing, laminar differences could underlie adaptations specific to microchioptera.

Patterns of olivocochlear axonal branches.

The olivocochlear (OC) pathway is the source of major feedback control of ascending acoustic information. Two main patterns of axonal branching are evident at this and other levels of the auditory pathway: long-distance collaterals and branches involved in feedback-control loops. Only a minority of OC neurons project to both cochlea, consistent with a role for the system in sound localization. OC branches to the ventral cochlear nucleus provide the anatomical substrate for a feedback control loop that could aid in modulating the intensity of acoustic information being conveyed to higher cortical levels while still dampening the overall intensity of signaling from the cochlea itself.

Areas of cat auditory cortex as defined by neurofilament proteins expressing SMI-32.

The monoclonal antibody SMI-32 was used to characterize and distinguish individual areas of cat auditory cortex. SMI-32 labels non-phosphorylated epitopes on the high- and medium-molecular weight subunits of neurofilament proteins in cortical pyramidal cells and dendritic trees with the most robust immunoreactivity in layers III and V. Auditory areas with unique patterns of immunoreactivity included: primary auditory cortex (AI), second auditory cortex (AII), dorsal zone (DZ), posterior auditory field (PAF), ventral posterior auditory field (VPAF), ventral auditory field (VAF), temporal cortex (T), insular cortex (IN), anterior auditory field (AAF), and the auditory field of the anterior ectosylvian sulcus (fAES). Unique patterns of labeling intensity, soma shape, soma size, layers of immunoreactivity, laminar distribution of dendritic arbors, and labeled cell density were identified. Features that were consistent in all areas included: layers I and IV neurons are immunonegative; nearly all immunoreactive cells are pyramidal; and immunoreactive neurons are always present in layer V. To quantify the results, the numbers of labeled cells and dendrites, as well as cell diameter, were collected and used as tools for identifying and differentiating areas. Quantification of the labeling patterns also established profiles for ten auditory areas/layers and their degree of immunoreactivity. Areal borders delineated by SMI-32 were highly correlated with tonotopically-defined areal boundaries. Overall, SMI-32 immunoreactivity can delineate ten areas of cat auditory cortex and demarcate topographic borders. The ability to distinguish auditory areas with SMI-32 is valuable for the identification of auditory cerebral areas in electrophysiological, anatomical, and/or behavioral investigations.

Connections of cat auditory cortex: I. Thalamocortical system.

Despite the functional importance of the medial geniculate body (MGB) in normal hearing, many aspects of its projections to auditory cortex are unknown. We analyzed the MGB projections to 13 auditory areas in the cat using two retrograde tracers to investigate thalamocortical nuclear origins, topography, convergence, and divergence. MGB divisions and auditory cortex areas were defined independently of the connectional results using architectonic, histochemical, and immunocytochemical criteria. Each auditory cortex area received a unique pattern of input from several MGB nuclei, and these patterns of input identify four groups of cortical areas distinguished by their putative functional affiliations: tonotopic, nontonotopic, multisensory, and limbic. Each family of areas received projections from a functionally related set of MGB nuclei; some nuclei project to only a few areas (e.g., the MGB ventral division to tonotopic areas), and others project to all areas (e.g., the medial division input to every auditory cortical area and to other regions). Projections to tonotopic areas had fewer nuclear origins than those to multisensory or limbic-affiliated fields. All projections were organized topographically, even those from nontonotopic nuclei. The few divergent neurons (mean: 2%) are consistent with a model of multiple segregated streams ascending to auditory cortex. The expanded cortical representation of MGB auditory, multisensory, and limbic affiliated streams appears to be a primary facet of forebrain auditory function. The emergence of several auditory cortex representations of characteristic frequency may be a functional multiplication of the more limited maps in the MGB. This expansion suggests emergent cortical roles consistent with the divergence of thalamocortical connections.

Connections of cat auditory cortex: II. Commissural system.

The commissural projections between 13 areas of cat auditory cortex (AC) were studied using retrograde tracers. Areal and laminar origins were characterized as part of a larger study of thalamic input and cortical origins of projections to each area. Cholera toxin beta subunit (CTbeta) and cholera toxin beta subunit gold-conjugate (CTbetaG) were injected separately within an area or in different areas in an experiment. The areas were identified independently with SMI-32, which revealed differences in neurofilament immunoreactivity in layers III, V, and VI. Each area received convergent AC input from 3 to 6 (mean, 5) contralateral areas. Most of the projections (>75%) were homotopic and from topographically organized loci in the corresponding area. Heterotopic projections (>1 mm beyond the main homotopic projection) constituted approximately 25% of the input. Layers III and V contained >95% of the commissural neurons. Commissural projection neurons were clustered in all areas. Commissural divergence, assessed by double labeling, was less than 3% in each area. This sparse axonal branching is consistent with the essentially homotopic connectivity of the commissural system. The many heterotopic origins represent unexpected commissural influences converging on an area. Areas more dorsal on the cortical convexity have commissural projections originating in layers III and V; more ventral areas favor layer III at the expense of layer V, to its near-total exclusion in some instances. Some areas have almost entirely layer III origins (temporal cortex and area AII), whereas others have a predominantly layer V input (anterior auditory field) or dual contributions from layers III and V (the dorsal auditory zone). A topographic distribution of commissural cells of origin is consistent with the order observed in thalamocortical and corticocortical projections, and which characterizes all extrinsic projection systems (commissural, corticocortical, and thalamocortical) in all AC areas. Thus, laminar as well as areal differences in projection origin distinguish the auditory cortical commissural system.

Connections of cat auditory cortex: III. Corticocortical system.

The mammalian auditory cortex (AC) is essential for computing the source and decoding the information contained in sound. Knowledge of AC corticocortical connections is modest other than in the primary auditory regions, nor is there an anatomical framework in the cat for understanding the patterns of connections among the many auditory areas. To address this issue we investigated cat AC connectivity in 13 auditory regions. Retrograde tracers were injected in the same area or in different areas to reveal the areal and laminar sources of convergent input to each region. Architectonic borders were established in Nissl and SMI-32 immunostained material. We assessed the topography, convergence, and divergence of the labeling. Intrinsic input constituted >50% of the projection cells in each area, and extrinsic inputs were strongest from functionally related areas. Each area received significant convergent ipsilateral input from several fields (5 to 8; mean 6). These varied in their laminar origin and projection density. Major extrinsic projections were preferentially from areas of the same functional type (tonotopic to tonotopic, nontonotopic to nontonotopic, limbic-related to limbic-related, multisensory-to-multisensory), while smaller projections link areas belonging to different groups. Branched projections between areas were <2% with deposits of two tracers in an area or in different areas. All extrinsic projections to each area were highly and equally topographic and clustered. Intrinsic input arose from all layers except layer I, and extrinsic input had unique, area-specific infragranular and supragranular origins. The many areal and laminar sources of input may contribute to the complexity of physiological responses in AC and suggest that many projections of modest size converge within each area rather than a simpler area-to-area serial or hierarchical pattern of corticocortical connectivity.

Auditory cortex mapmaking: principles, projections, and plasticity.

Maps of sensory receptor epithelia and computed features of the sensory environment are common elements of auditory, visual, and somatic sensory representations from the periphery to the cerebral cortex. Maps enhance the understanding of normal neural organization and its modification by pathology and experience. They underlie the derivation of the computational principles that govern sensory processing and the generation of perception. Despite their intuitive explanatory power, the functions of and rules for organizing maps and their plasticity are not well understood. Some puzzles of auditory cortical map organization are that few complete receptor maps are available and that even fewer computational maps are known beyond primary cortical areas. Neuroanatomical evidence suggests equally organized connectional patterns throughout the cortical hierarchy that might underlie map stability. Here, we consider the implications of auditory cortical map organization and its plasticity and evaluate the complementary role of maps in representation and computation from an auditory perspective.

The distributed auditory cortex.

A synthesis of cat auditory cortex (AC) organization is presented in which the extrinsic and intrinsic connections interact to derive a unified profile of the auditory stream and use it to direct and modify cortical and subcortical information flow. Thus, the thalamocortical input provides essential sensory information about peripheral stimulus events, which AC redirects locally for feature extraction, and then conveys to parallel auditory, multisensory, premotor, limbic, and cognitive centers for further analysis. The corticofugal output influences areas as remote as the pons and the cochlear nucleus, structures whose effects upon AC are entirely indirect, and it has diverse roles in the transmission of information through the medial geniculate body and inferior colliculus. The distributed AC is thus construed as a functional network in which the auditory percept is assembled for subsequent redistribution in sensory, premotor, and cognitive streams contingent on the derived interpretation of the acoustic events. The confluence of auditory and multisensory streams likely precedes cognitive processing of sound. The distributed AC constitutes the largest and arguably the most complete representation of the auditory world. Many facets of this scheme may apply in rodent and primate AC as well. We propose that the distributed auditory cortex contributes to local processing regimes in regions as disparate as the frontal pole and the cochlear nucleus to construct the acoustic percept.

Focal projections of cat auditory cortex to the pontine nuclei.

The pontine nuclei (PN) receive projections from the auditory cortex (AC) and they are a major source of mossy fibers to the cerebellum. However, they have not been studied in detail using sensitive neuroanatomical tracers, and whether all AC areas contribute to the corticopontine (CP) system is unknown. We characterized the projection patterns of 11 AC areas with WGA-HRP. We also compared them with their corticothalamic and corticocollicular counterparts. A third objective was to analyze the structure of the CP axons and their terminals with BDA. Both tracers confirm that all AC areas projected to lateral, central, and medial ipsilateral pontine divisions. The strongest CP projections were from nontonotopic and polymodal association areas. Preterminal fibers formed single terminal fields having many boutons en passant as well as terminal endings, and there was a specific morphological pattern for each pontine target, irrespective of their areal origin. Thus, axons in the medial division had a simpler terminal architecture (type 1 terminal plexus); both the central and lateral pons received more complex endings (type 2 terminal plexus). Auditory CP topographical distribution resembled visual and somatosensory CP projections, which preserve retinotopy and somatotopy in the pons, respectively. However, the absence of pontine tonotopy suggests that the AC projection topography is unrelated to tonotopy. CP input to the medial and central pons coincides with the somatosensory and visual cortical inputs, respectively, and such overlap might subserve convergence in the cerebellum. In contrast, lateral pontine input may be exclusively auditory.

Decoding the auditory corticofugal systems.

The status of the organization of the auditory corticofugal systems is summarized. These are among the largest pathways in the brain, with descending connections to auditory and non-auditory thalamic, midbrain, and medullary regions. Auditory corticofugal influence thus reaches sites immediately presynaptic to the cortex, sites remote from the cortex, as in periolivary regions that may have a centrifugal role, and to the cochlear nucleus, which could influence early central events in hearing. Other targets include the striatum (possible premotor functions), the amygdala and central gray (prospective limbic and motivational roles), and the pontine nuclei (for precerebellar control). The size, specificity, laminar origins, and morphologic diversity of auditory corticofugal axons is consonant with an interpretation of multiple roles in parallel descending systems.

Decoding the auditory corticofugal systems.

The status of the organization of the auditory corticofugal systems is summarized. These are among the largest pathways in the brain, with descending connections to auditory and non-auditory thalamic, midbrain, and medullary regions. Auditory corticofugal influence thus reaches sites immediately presynaptic to the cortex, sites remote from the cortex, as in perolivary regions that may have a centrifugal role, and to the cochlear nucleus, which could influence early central events in hearing. Other targets include the striatum (possible premotor functions), the amygdala and central gray (prospective limbic and motivational roles), and the pontine nuclei (for precerebellar control). The size, specificity, laminar origins, and morphologic diversity of auditory corticofugal axons is consonant with an interpretation of multiple roles in parallel descending systems.

Auditory thalamocortical transformation: structure and function.

Communicative, predatory, and reproductive behaviors rely on the auditory thalamocortical system, a key nexus that combines, transforms, and distributes virtually all acoustic information relevant to survival. The rules of connectivity for this complex network, both anatomically and functionally, are only beginning to be uncovered. Although the auditory thalamocortical system shares many features with other modalities, its connectivity and information processing principles differ from those of other modalities in many ways. Some physiological and anatomical bases for these differences are the subject of this review.

Principles governing auditory cortex connections.

Topographic maps are common constituents of the primary auditory, visual, and somatic sensory cortex. However, in most cortical areas, no such maps have yet been identified, posing a conceptual problem for theories of cortical function centered on topography. What principle guides the organization of these other areas? We investigated this issue in cat auditory cortex. The connectional topography of five tonotopic areas and eight non-tonotopic areas was assessed using retrograde tract tracing and quantified by three metrics: clustering, dispersion, and separation. Clustering measures the spatial density of labeled neurons, dispersion provides an index of their spread, and separation serves as a scaling metric. These parameters each show that all auditory cortical regions receive precise and equally topographic connections from thalamic, corticocortical, and commissural sources. This isotropic principle suggests a common substrate for coordinating communication across the cortex and may reflect common mechanisms related to the developmental patterning of connections. This unifying principle extends to auditory and prefrontal cortex, and perhaps to other neocortical areas.

Concurrent tonotopic processing streams in auditory cortex.

The basis for multiple representations of equivalent frequency ranges in auditory cortex was studied with physiological and anatomical methods. Our goal was to trace the convergence of thalamic, commissural, and corticocortical information upon two tonotopic fields in the cat, the primary auditory cortex (AI) and the anterior auditory field (AAF). Both fields are among the first cortical levels of processing. After neurophysiological mapping of characteristic frequency, we injected different retrograde tracers at separate, frequency-matched loci in AI and AAF. We found differences in their projections that support the notion of largely segregated parallel processing streams in the auditory thalamus and cerebral cortex. In each field, ipsilateral cortical input amounts to approximately 70% of the number of cells projecting to an isofrequency domain, while commissural and thalamic sources are each approximately 15%. Labeled thalamic and cortical neurons were concentrated in tonotopically predicted regions and in smaller loci far from their spectrally predicted positions. The few double-labeled thalamic neurons (<2%) are consistent with the hypothesis that information to AI and AAF travels along independent processing streams despite widespread regional overlap of thalamic input sources. Double labeling is also sparse in both the corticocortical and commissural systems ( approximately 1%), confirming their independence. The segregation of frequency-specific channels within thalamic and cortical systems is consistent with a model of parallel processing in auditory cortex. The global convergence of cells outside the targeted frequency domain in AI and AAF could contribute to context-dependent processing and to intracortical plasticity and reorganization.