Biomedical subjects

David K Ryugo

Publications and source records attributed to David K Ryugo.

At least 19 recordsLinked to original sources

PubMed2006

Synaptic alterations at inner hair cells precede spiral ganglion cell loss in aging C57BL/6J mice.

Hearing deficits have often been associated with loss of or damage to receptor hair cells and/or degeneration of spiral ganglion cells. There are, however, some physiological abnormalities that are not reliably attributed to loss of these cells. The afferent synapse between radial fibers of spiral ganglion neurons and inner hair cells (IHCs) emerges as another site that could be involved in transmission abnormalities. We tested the hypothesis that the structure of these afferent terminals would differ between young animals and older animals with significant hearing loss. Afferent endings and their synapses were examined by transmission electron microscopy at approximately 45% distance from the basal end of the cochlea in 2-3 month-old and 8-12 month-old C57BL/6J mice. The number of terminals in older animals was reduced by half compared to younger animals. In contrast, there was no difference in the density of SGCs between the age groups. Older animals featured enlarged terminals and mitochondria and enlarged postsynaptic densities and presynaptic bodies. These morphological changes may be a combination of pathologic, adaptive and compensatory responses to sensory dysfunction. Improved knowledge of these processes is necessary to understand the role of afferent connectivity in dysfunction of the aging cochlea.

Aging↗

PubMed2006

Projections of the second cervical dorsal root ganglion to the cochlear nucleus in rats.

Physiological, anatomical, and clinical data have demonstrated interactions between somatosensory and auditory brainstem structures. Spinal nerve projections influence auditory responses, although the nature of the pathway(s) is not known. To address this issue, we injected biotinylated dextran amine into the cochlear nucleus or dorsal root ganglion (DRG) at the second cervical segment (C2). Cochlear nucleus injections retrogradely labeled small ganglion cells in C2 DRG. C2 DRG injections produced anterograde labeling in the external cuneate nucleus, cuneate nucleus, nucleus X, central cervical nucleus, dorsal horn of upper cervical spinal segments, and cochlear nucleus. The terminal field in the cochlear nucleus was concentrated in the subpeduncular corner and lamina of the granule cell domain, where endings of various size and shapes appeared. Examination under an electron microscope revealed that the C2 DRG terminals contained numerous round synaptic vesicles and formed asymmetric synapses, implying depolarizing influences on the target cell. Labeled endings synapsed with the stalk of the primary dendrite of unipolar brush cells, distal dendrites of presumptive granule cells, and endings containing pleomorphic synaptic vesicles. These primary somatosensory projections contribute to circuits that are hypothesized to mediate integrative functions of hearing.

Acoustic Stimulation↗

PubMed2006

Structural and functional classes of multipolar cells in the ventral cochlear nucleus.

Multipolar cells in the ventral cochlear nucleus (VCN) are a structurally and functionally diverse group of projection neurons. Understanding their role in the ascending pathway involves partitioning multipolar cells into distinct populations and determining where in the brain each sends its coded messages. In this study, we used retrograde labeling techniques in rats to identify multipolar neurons that project their axons to the ipsilateral dorsal cochlear nucleus (DCN), the contralateral CN, or both structures. Three rats received injections of biotinylated dextran amine in the ipsilateral DCN and diamidino yellow in the contralateral CN. Several radiate multipolar neurons (defined by their axonal projections to the ipsilateral DCN and their dendrites that traverse VCN isofrequency sheets) were double-labeled but over 70% were not. This result suggests two distinct populations: (1) radiate-commissural (RC) multipolar cells that project to the ipsilateral DCN and the contralateral CN, and (2) radiate multipolar cells that project exclusively (in this context) to the ipsilateral DCN. In a different group of animals, we retrogradely labeled multipolar neurons that project their axons to the contralateral CN and measured the size of their cell bodies. The mean size of this population (266 +/- 156 microm2) was significantly smaller than those of RC-multipolar cells (418 +/- 140 microm2). We conclude that the CN commissural pathway is composed of at least two components: (1) RC multipolar cells and (2) commissural multipolar cells that are small- and medium-sized neurons that project exclusively (in this context) to the contralateral CN. These results identify separate structural groups of multipolar cells that may correspond to physiological unit types described in the literature. They also provide protocols for isolating and studying different populations of multipolar cells to determine the neural mechanisms that govern their responses to sound.

Amidines↗

PubMed2006

Projections from auditory cortex to cochlear nucleus: A comparative analysis of rat and mouse.

Mammalian hearing is a complex special sense that involves detection, localization, and identification of the auditory stimulus. The cerebral cortex may subserve higher auditory processes by providing direct modulatory cortical projections to the auditory brainstem. To support the hypothesis that corticofugal projections are a conserved feature in the mammalian brain, this article reviews features of the rat corticofugal pathway and presents new data supporting the presence of similar projections in the mouse. The mouse auditory cortex was localized with electrophysiological recording and neuronal tracers were injected into AI. The cochlear nucleus was dissected and examined for terminal fibers by light and electron microscopy. Bouton endings were found bilaterally forming synapses with dendrites of granule cells of the cochlear nucleus. This report provides evidence for direct auditory cortex projections to the cochlear nucleus in the mouse. The distribution of projections to the granule cell domain and the synapses onto granule cell dendrites are consistent with what has been reported for rats and guinea pigs. These findings suggest a general plan for corticofugal modulation of ascending auditory information in mammals. Corticobulbar inputs to the auditory brainstem likely provided a survival advantage by improving sound detection and identification, thus allowing the development of complex social behaviors and the navigation of varied environments.

Animals↗

PubMed2005

Efficient quantification of afferent cochlear ultrastructure using design-based stereology.

The afferent synapse between the auditory nerve fiber and the inner hair cell (IHC) represents a critical junction for hearing. Elucidation of the structure at this site will help establish the substrate for normal sound encoding as well as pathologic processes associated with hearing dysfunction. Previous applications of unbiased (design-based) stereological principles have expanded our knowledge of neuro-morphological changes evident with the light microscope. Applying these principles at the level of the synapse is a promising morphometric approach for the efficient sampling of large reference spaces with electron microscopy. This study tests the accuracy of using ultra-thin sections at a fixed interval, known as disector pairs, to quantify afferent innervation density. We analyzed the total numbers of afferent terminals, synaptic thickenings, and synaptic bodies associated with each IHC in the C57BL/6J mouse cochlea, and confirmed the accuracy of the stereological approach in comparison to three-dimensional reconstructions of serial alternate sections. The higher sampling efficiency of the disector pair method rapidly increases precision while also reducing the largest source of variability, inter-animal differences. We conclude that ultrastructural quantification of afferent innervation can be accomplished in the cochlea using efficient design-based stereology.

Animals↗

PubMed2005

A modified Golgi staining protocol for use in the human brain stem and cerebellum.

The Golgi silver-impregnation method established itself as an important technique for distinguishing morphology at the individual neuron level. This technique has been especially useful for studying human neuroanatomy because it works on postmortem tissue but it is also unreliable and capricious. In this report, we describe a simple technique that was applied to human autopsy and tissue-bank material yielding useful results for the study of neuronal morphology in the brain stem and cerebellum. Human adult brain stems had been immersion-fixed in formalin for a period of time ranging from weeks to months. Brain stem tissue was cross-sectioned into 3-5mm thick slabs, centered about the cochlear nucleus. Slabs were processed under continuous vacuum (22-26 in. of Hg), a procedure that promoted penetration of reagents into the tissue. Tissue was sectioned using a Vibratome and mounted for light microscopy. The results demonstrated improved staining of neurons in the brain stem. Staining of the large synaptic endings of auditory nerve fibers called end bulbs of Held in the cochlear nucleus was especially evident. These results suggest that an age-graded series could be conducted to describe the development of these large auditory endings in humans.

Auditory Pathways↗

PubMed2005

Projections from the spinal trigeminal nucleus to the cochlear nucleus in the rat.

The integration of information across sensory modalities enables sound to be processed in the context of position, movement, and object identity. Inputs to the granule cell domain (GCD) of the cochlear nucleus have been shown to arise from somatosensory brain stem structures, but the nature of the projection from the spinal trigeminal nucleus is unknown. In the present study, we labeled spinal trigeminal neurons projecting to the cochlear nucleus using the retrograde tracer, Fast Blue, and mapped their distribution. In a second set of experiments, we injected the anterograde tracer biotinylated dextran amine into the spinal trigeminal nucleus and studied the resulting anterograde projections with light and electron microscopy. Spinal trigeminal neurons were distributed primarily in pars caudalis and interpolaris and provided inputs to the cochlear nucleus. Their axons gave rise to small (1-3 microm in diameter) en passant swellings and terminal boutons in the GCD and deep layers of the dorsal cochlear nucleus. Less frequently, larger (3-15 microm in diameter) lobulated endings known as mossy fibers were distributed within the GCD. Ventrally placed injections had an additional projection into the anteroventral cochlear nucleus, whereas dorsally placed injections had an additional projection into the posteroventral cochlear nucleus. All endings were filled with round synaptic vesicles and formed asymmetric specializations with postsynaptic targets, implying that they are excitatory in nature. The postsynaptic targets of these terminals included dendrites of granule cells. These projections provide a structural substrate for somatosensory information to influence auditory processing at the earliest level of the central auditory pathways.

Animals↗

PubMed2004

Two types of afferent terminals innervate cochlear inner hair cells in C57BL/6J mice.

Afferent synapses on inner hair cells (IHC) transfer auditory information to the central nervous system (CNS). Despite the importance of these synapses for normal hearing, their response to cochlear disease and dysfunction is not well understood. The C57BL/6J mouse is a model for presbycusis and noise-induced hearing loss because of its age-related hearing loss and susceptibility to acoustic over-exposure. In this context, we sought to establish normal synaptic structure in order to better evaluate synaptic changes due to presbycusis and noise exposure. Ultrastructural analysis of IHCs and afferent terminals was performed in a normal hearing 3-month-old C57BL/6J mouse at cochlear sites corresponding to 8, 16 and 32 kHz using semi-serial sections. A stereologic survey of random sections was conducted of IHCs in 11 additional mice. Two morphologically distinct groups of afferent terminals were identified at all 3 frequency locations in 11 out of 12 animals. "Simple" endings demonstrated classic features of bouton terminals, whereas "folded" endings were larger in size and exhibited a novel morphologic feature that consisted of a fully internalized double membrane that partially divided the terminal into two compartments. In many cases, the double membrane was continuous with the outer terminal membrane as if produced by an invagination. We still must determine the generality of these observations with respect to other mouse strains.

Afferent Pathways↗

PubMed2004

An animal model for cochlear implants.

OBJECTIVE: To test the feasibility of using the deaf white cat model of early-onset deafness. We studied the neuronal effects of prosthetic intervention with a clinical, "off-the-shelf" multichannel cochlear implant. METHODS: We placed cochlear implants in 5 deaf white kittens at age 12 and 24 weeks. The devices were activated and stimulated in the laboratory using a clinical speech processor programmed with a high-resolution continuous interleaved sampling (CIS) strategy for 8 to 24 weeks. Stimulus parameters were guided by electrically evoked brainstem responses and intracochlear-evoked potentials. Kittens were assessed with respect to their tolerance and general behavior in response to speech, music, and environmental sounds. RESULTS: Surgical complications were minimal, and kittens tolerated the experimental procedures well. Subjects were able to detect and respond to a specific sound played from a computer speaker. Electrophysiologic responses were reliably attainable and showed consistency with observed behavioral responses to sound. This experimental paradigm, using clinical devices, can be used in a practical research setting in cats. CONCLUSIONS: Deafness and other variations in neural activity result in many distinct changes to the central auditory pathways. Animal models will facilitate assessment of the reversibility of deafness-associated changes at the level of the neuron and its connections. Our observations of the feasibility of using clinical devices in animal models will enable us to simulate clinical conditions in addressing questions about the effects of "replacement" activity on the structure and function within the central auditory pathways in deafness.

Animals↗

PubMed2004

PHR1, a PH domain-containing protein expressed in primary sensory neurons.

Previously, we identified PHR1 as an abundantly expressed gene in photoreceptors and showed that it encodes four isoforms, each with N-terminal pleckstrin homology (PH) and C-terminal transmembrane domains. To better understand PHR1 function and expression, we made a Phr1 null mouse by inserting a beta-galactosidase/neor cassette into exon 3. In addition to photoreceptors, we found abundant expression of specific Phr1 splice forms in olfactory receptor neurons and vestibular and cochlear hair cells. We also found Phr1 expression in cells with a possible sensory function, including peripheral retinal ganglion cells, cochlear interdental cells, and neurons of the circumventricular organ. Despite this discrete expression in known and putative sensory neurons, mice lacking PHR1 do not have overt sensory deficits.

Amino Acid Sequence↗

PubMed2003

The source of corticocollicular and corticobulbar projections in area Te1 of the rat.

Cortical area Te1 in the rat commonly is associated with primary auditory cortex. It is the source of direct projections to the inferior colliculus (IC), superior olivary complex (SOC), and the cochlear nucleus (CN). A question that arises is whether these descending pathways derive from a common source or separate populations of cortical neurons. We addressed this question in seven rats by injecting either Diamidino yellow (DiY) or Fast blue (FB) into the IC and injecting the other tracer into the CN (n=4) or SOC (n=3). All injections were made on the left side of the brain. In a sample of sections through area Te1 in both hemispheres, we counted single- and double-labeled cells. We estimate that IC-projecting cells outnumber those projecting to the CN or SOC by at least a factor of ten. The source of corticofugal pathways to the left IC was heavily biased towards the same side of the brain (ipsi/contra ratio 8 +/- 2.5), whereas it was more equally distributed between the two hemispheres for the left CN and SOC (ipsi/contra ratios ranged from 0.7-2.3). Finally, we observed that only 10-20% of those cells filled with a tracer injection in the CN or SOC also contained the tracer injected into the IC. In a previous study, we observed a similarly small percentage of double labeled cells when FB and DiY were injected into the CN and SOC, respectively. Combined with the distinct laminar distribution of IC-, SOC-, and CN-projecting neurons within layer V, the results suggest that these three pathways largely derive from different populations of cortical neurons.

Amidines↗

PubMed2003

Multimodal inputs to the granule cell domain of the cochlear nucleus.

There is growing evidence that hearing involves the integration of many brain functions, including vision, balance, somatic sensation, learning and memory, and emotional state. Some of these integrative processes begin at the earliest stages of the central auditory system. In this review, we will discuss evidence that reveals multimodal projections into the granule cell domain of the cochlear nucleus.

Animals↗

PubMed2003

Discharge properties of identified cochlear nucleus neurons and auditory nerve fibers in response to repetitive electrical stimulation of the auditory nerve.

Using the in vitro isolated whole brain preparation of the guinea pig maintained at 29 degrees C, we intracellularly recorded and stained cochlear nucleus (CN) neurons and auditory nerve (AN) fibers. Discharge properties of CN cells and AN axons were tested in response to 50-ms trains of electrical pulses delivered to the AN at rates ranging from 100 to 1000 pulses per second (pps). At low stimulation rates (200-300 pps), the discharges of AN fibers and a large proportion of principal cells (bushy, octopus, stellate) in the ventral cochlear nucleus (VCN) followed with high probability each pulse in the train, resulting in synchronization of discharges within large populations of AN fibers and CN cells. In contrast, at high stimulation rates (500 pps and higher), AN fibers and many VCN cells exhibited "primary-like", "onset" and some other discharge patterns resembling those produced by natural sound stimuli. Unlike cells in the VCN, principal cells (pyramidal, giant) of the dorsal CN did not follow the stimulating pulses even at low rates. Instead, they often showed "pauser" and "build-up" patterns of activity, characteristic for these cells in conditions of normal hearing. We hypothesize that, at low stimulation rates, the response behavior of AN fibers and VCN cells is different from the patterns of neuronal activity related to normal auditory processing, whereas high stimulation rates produce more physiologically meaningful discharge patterns. The observed differences in discharge properties of AN fibers and CN cells at different stimulation rates can contribute to significant advantages of high- versus low-rate electrical stimulation of the AN used for coding sounds in modern cochlear implants.

Action Potentials↗

PubMed2003

Axonal pathways to the lateral superior olive labeled with biotinylated dextran amine injections in the dorsal cochlear nucleus of rats.

The lateral superior olive (LSO) contains cells that are sensitive to intensity differences between the two ears, a feature used by the brain to localize sounds in space. This report describes a source of input to the LSO that complements bushy cell projections from the ventral cochlear nucleus (VCN). Injections of biotinylated dextran amine (BDA) into the dorsal cochlear nucleus (DCN) of the rat label axons and swellings in several brainstem structures, including the ipsilateral LSO. Labeling in the ipsilateral LSO was confined to a thin band that extended throughout the length of the structure such that it resembled an LSO isofrequency lamina. The source of this labeled pathway was not obvious, because DCN neurons do not project to the LSO, and VCN bushy cells were not filled by these injections. Filled neurons in several brainstem structures emerged as possible sources. Three observations suggest that most of the axonal labeling in the LSO derives from a single source. First, the number of labeled VCN planar multipolar cells and the amount of labeling in the LSO were consistent and robust across animals. In contrast, the number of labeled cells in most other structures was small and highly variable. Second, the locations of planar cells and filled axons in the LSO were related topographically to the position of the DCN injection site. Third, labeled terminal arborizations in the LSO arose from collaterals of axons in the trapezoid body (output tract of planar cells). We infer that planar multipolar cells, in addition to bushy cells, are a source of ascending input from the cochlear nucleus to the LSO.

Animals↗

PubMed2003

Primary innervation of the avian and mammalian cochlear nucleus.

The auditory nerve of birds and mammals exhibits differences and similarities, but given the millions of years since the two classes diverged from a common ancestor, the similarities are much more impressive than the differences. The avian nerve is simpler than that of mammals, but share many fundamental features including principles of development, structure, and physiological properties. Moreover, the available evidence shows that the human auditory nerve follows this same general organizational plan. Equally impressive are reports that homologous genes in worms, flies, and mice exert the same heredity influences in man. The clear implication is that animal studies will produce knowledge that has a direct bearing on the human condition.

Animals↗

PubMed2003

Ultrastructural examination of the somatic innervation of ventrotubercular cells in the rat.

Ventrotubercular cells are multipolar cells in the ventral cochlear nucleus (VCN) that project a collateral axon to the ipsilateral dorsal cochlear nucleus (DCN). These cells are thought to be involved in sensitizing DCN output neurons to spectral shapes that represent the location of a sound source in space. The present report focused on the neuronal composition of this pathway. Intracellular labeling studies in cats and mice have described two types of ventrotubercular cells (Smith and Rhode [1989] J Comp Neurol. 282:595-626; Oertel et al. [1990] J Comp Neurol. 295:136-154). In cats, one difference between the two classes is that type I multipolar neurons have fewer than 35% of their somata apposed by terminals, whereas type II cells have greater than 70% apposition values. Intracellular recordings from single cells, however, are difficult and thus limit the yield of data. We investigated whether a two-component description of the ventrotubercular pathway was representative of a larger population. This issue was addressed by retrogradely labeling ventrotubercular neurons with an extracellular injection of biotinylated dextran amine into the DCN of rats. These injections labeled many VCN neurons, thus providing a more complete view of the pathway than previous studies. Thirty-eight labeled cells were selected for electron microscopic analysis with respect to their location, cell body size, and ultrastructural morphology. We observed labeled type I and type II neurons, but unlike ventrotubercular cells in cats, many of these neurons in rats (17 of 38 cells) had appositions between 35% and 70%. On the basis of this analysis, a third class of ventrotubercular cell, called the adendritic neuron, was revealed. Adendritic neurons have small somata with many filopodial appendages, no observable dendrites, and high percentage of terminal appositions (>80%). The results demonstrated that the ventrotubercular pathway in the rat is diverse.

Animals↗

PubMed2003

Separate forms of pathology in the cochlea of congenitally deaf white cats.

Congenital deafness due to cochlear pathology can have an immediate or progressive onset. The timing of this onset could have a significant impact on the development of structures in the central auditory system, depending on the animal's hearing status during its critical period. In order to determine whether cats in our deaf white cat colony suffered from progressive hearing loss, they were tested repeatedly in 30-day intervals using standard auditory evoked brainstem response (ABR) methodology. ABR thresholds did not change over time, indicating that deafness in our colony was not progressive. Moreover, different forms of cochlear pathology were associated with deafness. One form (67% of the deaf ears) had a collapsed Reissner's membrane that obliterated the scala media, resembling what is called the Scheibe deformity in humans. A second form (18%) exhibited excessive epithelial growth within the bony labyrinth. A third form (15%) combined excessive epithelial growth in the apex and a collapsed Reissner's membrane in the base. Cochleae having an abnormally thin tectorial membrane and an outward bulging Reissner's membrane were associated with elevated thresholds (poor hearing).

Aging↗

PubMed2003

The functional age of hearing loss in a mouse model of presbycusis. II. Neuroanatomical correlates.

This report relates patterns of age-related outer hair cell (OHC) loss to auditory behavioral deficits in C57BL/6J mice. Hair cell counts were made from serial sections of the cochlear partition in three subject groups representing young (2-3 months), middle (8-9 months), and old ages (12-13 months). The cochlear location of OHC counts was determined from three-dimensional computerized reconstructions of the serial sections. Comparisons of the topographic distribution of surviving OHCs across the subject groups confirmed an orderly base-to-apex progression of cochlear degeneration that is well known in this mouse strain. All mice appeared to follow the same progression of OHC loss, although subjects showed considerable variation in the rate at which they advanced through a uniform sequence of structural changes. Behavioral implications of the magnitude and location of OHC loss were investigated by correlating the histological status of individual mice with sound detection thresholds from the same subjects [Hear. Res. 183 (2003) 44-56]. The analysis revealed regionalized patterns of OHC loss that were correlated with frequency-dependent changes in hearing thresholds, and validates the use of 'functional age' as an indicator of age-related cochlear degeneration and dysfunction. In the absence of physiologically defined cochlear frequency maps for C57BL/6J mice, these structure-function correlation techniques offer an alternative approach for linking anatomical results to hearing abilities.

Age Factors↗