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PubMed1996

Expression of insulin receptor-related receptor in the rat brain examined by in situ hybridization and immunohistochemistry.

Insulin receptor-related receptor (IRR) is a member of the insulin receptor family. However, its endogenous ligand and physiological roles are unknown. To elucidate the physiological roles of IRR, an orphan receptor, in the brain, we examined its expression at mRNA and protein levels in the brain by in situ hybridization and immunohistochemistry, respectively. The expression of IRR mRNA in the brain was highly restricted to the forebrain including the nucleus of the diagonal band, medial septal nucleus, ventral pallidum, accumbens nucleus and caudate putamen, and the brainstem including the prepositus hypoglossal nucleus, medial vestibular nucleus, gigantocell reticular nucleus, paragigantocellular nucleus and ventral cochlear nucleus. Most IRR mRNA-positive cells in the forebrain but not in the brainstem were cholinergic neurons. However, most IRR mRNA in the forebrain and brainstem was coexpressed with that of trkA, a high-affinity receptor for nerve growth factor. IRR-immunoreactive cell bodies were also detected in the forebrain and brainstem. The pattern of IRR immunoreactivity was similar to that of IRR mRNA. Its restricted pattern indicates that IRR plays unique roles in the brain, in contrast to insulin and insulin-like growth factor-I receptors, other members of the insulin receptor family, which are widely expressed in the brain.

Animals↗
PubMed2000

Identification of a brain specific protein that associates with a refsum disease gene product, phytanoyl-CoA alpha-hydroxylase.

Refsum disease is an autosomal recessive neurologic disorder of the lipid metabolism. Major diagnostic clinical findings include retinitis pigmentosa, peripheral polyneuropathy, cerebellar ataxia, increased cerebrospinal fluid protein without pleocytosis, nerve deafness, and cardiac involvement. We have identified a novel protein (PAHX-AP #1) associated with phytanoyl-CoA alpha-hydroxylase (PAHX), a Refsum disease gene product, using the yeast-based two-hybrid assay. The middle portion (amino acids 83-264) of PAHX was used as a bait and a mouse brain cDNA library was searched. The ability of PAHX-AP #1 to interact with PAHX was confirmed using immunoprecipitation and Western blot studies in NIH3T3 cells which stably expressed both PAHX and PAHX-AP #1. Northern and Western blot analyses demonstrated a unique pattern of developmental PAHX-AP #1 expression which was targeted to the adult brain, but ubiquitous expressions of PAHX were observed in all examined tissues. In situ hybridization analyses of the brain showed specific localization of PAHX-AP #1 to the supragranular layer in the cerebral cortex, dentate gyrus, hippocampus, Purkinje cell layer, deep cerebellar nucleus, trigeminal nucleus, abducent nucleus, facial nucleus, cochlear and vestibular nucleus, ganglion cell and nuclear layer of the retina. These data indicate that localization of PAHX-AP #1 in the brain is correlated with central neurologic symptoms of Refsum disease such as retinitis pigmentosa, cerebellar ataxia, nerve deafness and suggest that PAHX-AP #1 may be involved in the development of the central neurologic deficits of Refsum disease.

3T3 Cells↗
PubMedDate not supplied

The auditory pathology of anoxia.

A case of anoxic encephalopathy is reported, with study on a whole-auditory-pathways basis, and the method of processing tissues is outlined. Pathologic changes are found throughout the central part of the auditory pathway. The importance of including the superior ventral cochlear nucleus (SVCN) with cochlear structures in correlating findings with audiometric data is supported. The previously advanced tonotopic frequency pattern, with zonal vulnerability, of the spheroid cells of SVCN is supported.

Auditory Cortex↗
PubMed1987

Gaze-evoked tinnitus.

Two patients developed gaze-evoked tinnitus in the year following removal of a cerebellopontine angle mass. The eighth nerve was transected in both cases. The noise was present with saccades, pursuit, and vestibuloocular eye movements. We postulate the tinnitus was due to an abnormal interaction between the vestibular nucleus and the cochlear nucleus, possibly secondary to neural sprouting.

Eye Movements↗
PubMed1999

Distribution of cytoplasmic phospholipase A2 in the normal rat brain.

The calcium-dependent cytoplasmic PLA2 (cPLA2) is an 85-kDa cytosolic enzyme that has been detected in cytosolic fractions from rat brain. With immunocytochemical methods, this cPLA2 is distributed throughout rat brain. Very dense immunostaining is observed in the superior olivary nucleus, periolivary nucleus, facial motor nucleus and dorsal cochlear nucleus in hindbrain whereas light immunostaining is seen in forebrain and midbrain areas. Assays of cPLA2 activity in forebrain, midbrain and hindbrain show the highest specific activity in the hindbrain. The distribution of cPLA2 coincides with that of protein kinase C activity in rat brain. The presence of cPLA2 and PKC in hindbrain suggests that these enzymes play a central role in neurotransmitter release, long-term potentiation and neuritogenesis in this area under normal conditions.

Animals↗
PubMed2000

Mapping of alpha-melanocyte-stimulating hormone-like immunoreactivity in the cat brainstem.

The distribution of alpha-melanocyte-stimulating hormone-like immunoreactive structures was studied in the brainstem of the cat using an indirect immunoperoxidase technique. Immunoreactivity was observed in several brainstem nuclei of the cat in which no immunoreactivity had been previously reported. Immunoreactive fibres were observed in the following; the inferior central nucleus; the pontine gray nuclei; the Kölliker-Fuse nucleus; the motor trigeminal nucleus, the anteroventral cochlear nucleus; the abducens nucleus; the retrofacial nucleus; the superior, lateral, inferior, and medial vestibular nuclei; the lateral nucleus of the superior olive; the external cuneate nucleus; the nucleus of the trapezoid body; the postpyramidal nucleus of the raphe; the medial accessory inferior olive; the dorsal accessory nucleus of the inferior olive; the nucleus ambiguus; the principal nucleus of the inferior olive; the preolivary nucleus; the nucleus ruber; the substantia nigra; and in the area postrema. Our results point to a more widespread distribution of alpha-melanocyte-stimulating hormone-like immunoreactive structures in the cat brainstem than that reported in previous studies carried out in the same region of the cat, rat and humans.

Animals↗
PubMed1994

Light and electron microscope distribution of the NMDA receptor subunit NMDAR1 in the rat nervous system using a selective anti-peptide antibody.

NMDA receptors play key roles in synaptic plasticity and neuronal development, and may be involved in learning, memory, and compensation following injury. A polyclonal antibody that recognizes four of seven splice variants of NMDAR1 was made using a C-terminus peptide (30 amino acid residues). NMDAR1 is the major NMDA receptor subunit, found in most or all NMDA receptor complexes. On immunoblots, this antibody labeled a single major band migrating at M(r) = 120,000. The antibody did not cross-react with extracts from transfected cells expressing other glutamate receptor subunits, nor did it label non-neuronal tissues. Immunostained vibratome sections of rat tissue showed labeling in many neurons in most structures in the brain, as well as in the cervical spinal cord, dorsal root and vestibular ganglia, and in pineal and pituitary glands. Staining was moderate to dense in the olfactory bulb, neocortex, striatum, some thalamic and hypothalamic nuclei, the colliculi, and many reticular, sensory, and motor neurons of the brainstem and spinal cord. The densest stained cells included the pyramidal and hilar neurons of the CA3 region of the hippocampus, Purkinje cells of the cerebellum, supraoptic and magnocellular paraventricular neurons of the hypothalamus, inferior olive, red nucleus, lateral reticular nucleus, peripheral dorsal cochlear nucleus, and motor nuclei of the lower brainstem and spinal cord. Ultrastructural localization of immunostaining was examined in the hippocampus, cerebral cortex, and cerebellar cortex. The major staining was in postsynaptic densities apposed by unstained presynaptic terminals with round or mainly round vesicles, and in associated dendrites. The pattern of staining matched that of previous in situ hybridization but differed somewhat from that of binding studies, implying that multiple types of NMDA receptors exist. Comparison with previous studies of localization of other glutamate receptor types revealed that NMDAR1 may colocalize with these other types in many neurons throughout the nervous system.

Alternative Splicing↗
PubMed1996

[Effects of chronic electrical stimulation on cochlear nuclear neuron size in deaf kittens].

It is now well recognized that normal afferent innervation is necessary for development of the auditory pathway. This study investigated the effect of chronic electrical stimulation of the auditory nerve on the size of cochlear nucleus somata of neonatally deafened kittens. Four kittens were deafened by using kanamycin and ethacrynic acid at 10 days of age and implanted at various ages with an active intracochlear implant in the left side and a dummy implant in the right side, and chronically electrically stimulated from ages 98-142 days for approximately 1000 hours at twice the EABR threshold. After chronic stimulation or after an equivalent period in the controls, the animals were injected with 2-deoxyglucose (2DG) and stimulated electrically for 45 minutes, then processed for autoradiography. Then the sections were stained for Nissle substance, and the cross-sectional areas (CSSA) of approximately 33,000 neuron somata in the cochlear nuclei were measured with an image-analysis system. Soma sizes in regions with 2DG uptake were compared with those without the uptake. The CSSA of 2DG-labeled regions was usually significantly larger than that of the unlabeled regions, but there were no significant right-left differences. Thus, chronic electrical stimulation of a restricted sector of the cochlea is correlated with a larger soma size in the corresponding region of the cochlear nucleus.

Animals↗
PubMed2002

Auditory system development: primary auditory neurons and their targets.

The neurons of the cochlear ganglion transmit acoustic information between the inner ear and the brain. These placodally derived neurons must produce a topographically precise pattern of connections in both the inner ear and the brain. In this review, we consider the current state of knowledge concerning the development of these neurons, their peripheral and central connections, and their influences on peripheral and central target cells. Relatively little is known about the cellular and molecular regulation of migration or the establishment of precise topographic connection to the hair cells or cochlear nucleus (CN) neurons. Studies of mice with neurotrophin deletions are beginning to yield increasing understanding of variations in ganglion cell survival and resulting innervation patterns, however. Finally, existing evidence suggests that while ganglion cells have little influence on the differentiation of their hair cell targets, quite the opposite is true in the brain. Ganglion cell innervation and synaptic activity are essential for normal development of neurons in the cochlear nucleus.

Animals↗
PubMed1997

Effect of adult-onset deafness on the human central auditory system.

Degenerative change in the central auditory system was assessed in seven subjects with profound bilateral adult-onset deafness. The degree of transneuronal atrophy was determined by measuring cell size at three levels of the brain stem auditory pathway (anteroventral cochlear nucleus, medial superior olivary nucleus, and inferior colliculus). Within subjects, the relative degree of cell shrinkage was similar across all levels of the central pathway. Across subjects, the best neuronal preservation was seen in a case of viral labyrinthitis with 1 year of bilateral dearness and a near-normal population of cochlear ganglion cells. Reduction in cell size was greatest in cases of bacterial labyrinthitis or Scheibe degeneration with reduced populations of ganglion cells and longer periods of deafness. At the level of the cochlear nucleus, there was no consistent difference in cell size between the side stimulated by a functioning prosthetic device and the nonstimulated side.

Adult↗
PubMed1993

Quantitative comparison of electrically and acoustically evoked auditory perception: implications for the location of perceptual mechanisms.

Electrical stimulation of the human auditory system produces different patterns of spatial and temporal neural activity than those that occur in the normal, acoustically stimulated system. Quantitative comparison of psychophysical performance measured with acoustic and electrical stimulation may allow us to infer the physiological locus of perceptual mechanisms. In this paper we compare psychophysical data on temporal resolution from normal-hearing listeners, cochlear implant listeners, and patients electrically stimulated on the cochlear nucleus. Measures of gap detection, forward masking, and modulation detection will be compared. These comparisons demonstrate that temporal processing is relatively similar across these three groups once the obvious differences in dynamic range are taken into consideration. In addition, preliminary results with speech processors indicate that implant patients can utilize all temporal information in speech. Thus, implant patients have relatively normal temporal resolution and can integrate temporal cues normally for the recognition of complex acoustic patterns such as speech. These results imply that the central auditory systems of implant patients are able to fully utilize the non-natural patterns of temporal neural information produced by electrical stimulation. The differences in the microstructure of the neural pattern (phase locking, stochastic independence of fibers, spatial distribution of activity, etc.) between electrical and acoustic stimulation are apparently not necessary for temporal processing. Thus, the physiological locus of temporal processing mechanisms must be more central in the auditory system than the cochlea and cochlear nucleus.

Acoustic Stimulation↗
PubMed1994

Neurotrophins affect survival and neuritogenesis by adult injured auditory neurons in vitro.

This study evaluates the trophic effects of three neurotrophins on traumatized adult auditory neurons in culture, and the presence of these neurotrophins in cochlear nucleus tissue. BDNF and NT-3 promoted survival but very limited neuritogenesis by adult auditory neurons in vitro, while NGF, although without a survival effect, evoked a robust neuritic outgrowth response when combined with BDNF. Messenger RNAs that encode for NGF, BDNF and NT-3 were detected by RT-PCR in RNA extracts from adult cochlear nuclei tissue. Based on these in vitro and in vivo findings, we propose NT-3 as the agent of the peripheral target-derived survival promoting effect and NGF, BDNF, and NT-3 as mediators of trophic influences originating from the central target (i.e. cochlear nucleus).

Animals↗
PubMed1999

Inhibitory synaptic interactions between cochlear nuclei: evidence from an in vitro whole brain study.

Using guinea-pig isolated whole brain preparation in vitro, synaptic responses to electrical stimulation of auditory nerves were examined in intracellularly recorded and stained neurons of posteroventral and dorsal divisions of the cochlear nucleus. Stimulation of the contralateral auditory nerve evoked exclusively IPSPs in 70% of neurons, with amplitude of 2.3+/-1.2mV. Neurons of all major cell types were inhibited from the contralateral side. In the majority of responding cells (78%) IPSPs were induced at latencies of 3-9 ms suggesting di- and trisynaptic connections from contralateral auditory afferents or, respectively, mono- and disynaptic connections from the contralateral cochlear nucleus. Few cells responded with long-latency IPSPs (13.5-23ms), indicating involvement of polysynaptic pathways. These data demonstrate the existence of functional, direct and indirect inhibitory connections between the cochlear nuclei.

Animals↗
PubMed1996

Distribution of calcium-binding protein immunoreactivities in the guinea pig auditory brainstem.

This study was intended to provide an overview of the distribution of calcium-binding proteins in the rodent auditory brainstem. We based our observations on immunohistochemical material obtained in the guinea pig, a species widely used in auditory research in which a mapping of calcium-binding proteins in the auditory brainstem is still missing. Differences in the amounts of these proteins throughout the auditory brainstem were further analyzed semiquantitatively. Parvalbumin was present in most neurons and their axon terminals throughout the ascending auditory brainstem. Nuclei that surround the main relay nuclei of the ascending auditory pathway lacked labeling. Calretinin staining was prominent in spherical and globular cells of the cochlear nucleus, in their axon terminals in the superior olivary complex, and in principal cells of the medial superior olive. Measures of optical densities showed that auditory neurons involved in sound localization had the highest calretinin labeling levels. Calbindin D-28k was present in cartwheel cells of the dorsal cochlear nucleus, in almost all neurons of the medial nucleus of the trapezoid body, and in globular cells in the ventral nucleus of the lateral lemniscus. The labeling patterns for calretinin and calbindin D-28k were non-overlapping throughout the auditory brainstem. This was also evident in the ventral nucleus of the lateral lemniscus where calbindin D-28k-immunoreactive terminals were found in the medial portion, while the calretinin-immunoreactive terminals were observed in the lateral portion. This study presents the first direct and comprehensive comparison of these three calcium-binding proteins in the auditory brainstem of a rodent. Each antibody yields a unique staining pattern that provides a basis for further defining neuronal populations. In addition, since their axons are also selectively stained, auditory nuclei can further be compartmentalized based on different terminal fields. These immunoreactivities have provided clues to the complex structure of the auditory brainstem.

Animals↗
PubMed2001

Encoding of the temporal regularity of sound in the human brainstem.

We measured the neural activity associated with the temporal structure of sound in the human auditory pathway from cochlear nucleus to cortex. The temporal structure includes regularities at the millisecond level and pitch sequences at the hundreds-of-milliseconds level. Functional magnetic resonance imaging (fMRI) of the whole brain with cardiac triggering allowed simultaneous observation of activity in the brainstem, thalamus and cerebrum. This work shows that the process of recoding temporal patterns into a more stable form begins as early as the cochlear nucleus and continues up to auditory cortex.

Acoustic Stimulation↗
PubMed1996

gamma-Aminobutyric acid and glycine in the baboon cochlear nuclei: an immunocytochemical colocalization study with reference to interspecies differences in inhibitory systems.

Previous studies of the cochlear nuclei in cat, rat, and guinea pig have demonstrated neural structures that are enriched in the inhibitory neurotransmitter amino acids gamma-aminobutyric acid (GABA) and glycine. In these mammals, inhibitory terminals are widely distributed throughout the nuclear complex, but somata of inhibitory neurons are concentrated in the dorsal cochlear nucleus, in granule cell regions, and in the cap area. Because these are the subdivisions that undergo the most pronounced phylogenetic changes in primates, we wanted to see whether the inhibitory systems are influenced by changes in cytoarchitecture. Therefore, we applied light microscopic postembedding immunostaining and optical densitometry to the cochlear nuclei of an anthropoid primate, the Senegalese baboon (Papio anubis). Our results demonstrate that, in baboon 1) glycinergic neurons and axons in the ventral cochlear nucleus seem to form a commissural system similar to that of other mammals; 2) the tuberculoventral system appears to be unchanged in morphology but exhibits a higher level of colocalization of GABA with glycine; 3) there is a reduction of the granule/cartwheel cell system, which is reflected in lesser numbers of inhibitory cartwheel, Golgi, and molecular layer stellate cells; 4) the cap area is larger than in rodents and carnivores and contains many neurons that colocalize GABA and glycine; and 5) throughout the nuclear complex, a higher proportion of the inhibitory terminals colocalize GABA and glycine. We conclude that modulation of the ascending auditory pathway in baboon is likely to differ from that in rodents and cat.

Animals↗
PubMed2001

In vivo visualization of the cochlear nerve and nuclei with fluorescent axonal tracers.

In recent years multichannel neuroprostheses have been developed which directly stimulate the central auditory pathway. Substantially these have been used in cases of total hearing loss caused by neurofibromatosis type 2 where bilateral damage to the auditory nerve prevents more peripheral stimulation. The electrode carrier of the auditory brainstem implant (ABI) is designed to be placed on the cochlear nucleus complex residing at the lateral brainstem surface. Despite altered anatomy due to tumor growth or preceding surgery, correct electrode placement is essential to maximize the variety of pitch percept elicited during electrical stimulation with the ABI without producing side-effects. In order to assist intraoperative identification of the proximal auditory nerve and cochlear nuclei, the non-toxic fluorescent axonal tracers Fast Blue or Fluorogold were injected into the cochlea of rats and Java monkeys. Four to seven days after tracer application, labeling of the eighth cranial nerve, its entrance into the brainstem and the primary radiation of auditory fibers into the cochlear nucleus could be demonstrated as colored fluorescence on the living brain under appropriate ultraviolet illumination. Additional histological processing revealed groups of retrogradely labeled neuronal cell bodies in both species. Our results suggest that this method could also be used in humans in order to aid surgeons with the proper positioning of the electrode array.

Amidines↗
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↗