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J Fex

Publications and source records attributed to J Fex.

At least 37 records · Page 2Linked to original sources
PubMed1986

Neurotransmitter-related immunocytochemistry of the organ of Corti.

The principles of immunocytochemistry were outlined in 1942 by Coons et al. and in the 1970's immunocytochemistry emerged as a powerful method for identifying structures and tracing pathways in the nervous system. It now plays a fundamental role in the neuroanatomical and histochemical analysis of the central nervous system. The first immunocytochemical studies of the mammalian cochlea were reported in 1980, from three different laboratories. Since then many studies on cochlear immunocytochemistry have been carried out, concerned with questions about neurotransmitter candidates or about structural proteins. This review describes immunoreactivity of enkephalin, choline acetyltransferase (ChAT), glutamate decarboxylase (GAD), gamma-aminobutyric acid (GABA), aspartate aminotransferase (AATase) and glutaminase (GLNase) in the organ of Corti. ChAT is the enzyme that catalyzes the synthesis of acetylcholine (ACh). GAD is the terminal enzyme in the biosynthesis of the inhibitory neurotransmitter GABA. AATase and GLNase are two enzymes involved in the metabolism of the excitatory neurotransmitter candidates aspartate and glutamate. We have much relied on surface preparations of the organ of Corti. We have also used cryostat sectioning of the cochlea, particularly when there was a need to apply a number of different antisera to comparable preparations from one and the same cochlea. We have used immunofluorescence and immunoperoxidase procedures. Immunoperoxidase procedures have given us better signal noise ratio for specific immunoreactivity (in surface preparations) than has immunofluorescence. Occasionally, to achieve maximal resolution of surface preparations in light microscopy studies, we have used enhanced contrast video display. We have found immunoreactivity in efferent fibers in the organ of Corti following the application of antisera to enkephalin, ChAT, GAD, GABA, AATase and GLNase. Most of these different antisera give different distributions of immunoreactivity and other antisera have evoked no immunoreactivity in the organ of Corti. To the best of our knowledge, the cells of origin of efferent axons and terminals in the organ of Corti are located in the brainstem. Originally described as crossed and uncrossed olivocochlear neurons, these efferents have recently been classified into a medial and a lateral system predominantly innervating, respectively, the outer hair cell region and the inner hair cell region. However, our findings on the distribution of GAD- and GABA-like immunoreactivity indicate that there may be more than two different systems of efferents in the organ of Corti, as previously suggested by Schwartz and Ryan (1983).

Animals↗
PubMed1985

Immunocytochemical localization of choline acetyltransferase-like immunoreactivity in the guinea pig cochlea.

The immunocytochemical localization of the enzyme choline acetyltransferase (ChAT) was examined in the guinea pig organ of Corti to determine if both lateral and medial systems of efferents would show immunoreactive labeling for this specific enzyme marker of cholinergic neurons. Cochleae were also examined after lesion of efferents to determine if ChAT-like immunoreactivity is confined to efferents. ChAT-like immunoreactivity was seen in the inner spiral bundle, tunnel spiral bundle and by the bases of inner hair cells corresponding to the lateral system of efferents. ChAT-like immunoreactivity was also seen in crossing fibers and puncta at the bases and by the nuclei of outer hair cells corresponding to the medial system of efferents. With the use of video enhanced contrast microscopy more than 9 ChAT-like immunoreactive puncta at the bases of outer hair cells could be resolved. In cochleae examined 6 weeks after ipsilateral lesion of efferents, no ChAT-like immunoreactivity was observed. These results add strong evidence that acetylcholine is a transmitter of both the medial and lateral systems of efferents.

Acetylcholine↗
PubMed1985

Neuron-specific enolase-like immunoreactivity in inner hair cells but not outer hair cells in the guinea pig organ of Corti.

Neuron-specific enolase (NSE) has been localized only in neurons and cells with characteristics of neurons. The immunocytochemical localization of NSE was examined in guinea pig cochleae to determine if hair cells, which have some neuronal characteristics, would show NSE-like immunoreactive labeling. NSE-like immunoreactivity was seen in inner hair cells but not in outer hair cells. This is the first report of NSE-like immunoreactivity in a receptor cell. NSE-like immunoreactivity was also seen in efferent fibers and terminals and in both type I and type II spiral ganglion cells. The finding of NSE-like immunoreactivity in inner but not outer cells adds to the number of differences found between them and may be related to differences in function and action.

Animals↗
PubMed1985

Glutaminase-like immunoreactivity in the organ of Corti of guinea pig.

The distribution of glutaminase (GLNase)-like immunoreactivity (IR) in the normal and surgically de-efferented organ of Corti of guinea pig was studied. Primary antisera were against phosphate-dependent GLNase from rat kidney. Indirect immunocytochemical techniques were used; IR was visualized in cryostat sections through immunofluorescence, and through immunofluorescence or with horseradish peroxidase reaction product in surface preparations. Standard microscopy and video-enhanced light microscopy with asymmetric illumination contrast were used. GLNase-like IR was found at inner hair cells (IHCs) in the normal and in the de-efferented organ of Corti, in the tunnel spiral bundle, in tunnel-crossing fibers, in endings high up on outer hair cells (OHCs), in outer spiral bundles, in puncta close to OHCs, and in large, efferent endings at OHC bases. There was no GLNase-like IR at OHCs in the de-efferented organ of Corti. It is concluded that GLNase-like IR is present in auditory nerve dendrites at IHCs and in olivocochlear efferents of the medial system, and that future studies are needed to determine whether also the lateral system of olivocochlear efferents contains GLNase-like IR. A diagram is included depicting the relation between OHCs and efferent nerve endings along the cochlear spiral, showing that in the apicalmost 3/4 turn of the spiral OHCs have no efferent endings.

Animals↗
PubMed1985

Localization of dynorphin B-like and alpha-neoendorphin-like immunoreactivities in the guinea pig organ of Corti.

Antiserum to dynorphin B and antiserum to alpha-neoendorphin were used in an immunocytochemical examination of the guinea pig organ of Corti. Immunoreactive staining for these two proenkephalin B (prodynorphin)-derived peptides was seen in the lateral system of olivocochlear efferents in the organ of Corti: the inner spiral bundle, the tunnel spiral bundle and by the bases of inner hair cells. Immunoreactive staining with both antisera was also seen in efferent terminals on outer hair cells at or above the level of the nucleus, which may represent terminals of either the lateral or the medial system. No immunoreactive staining was seen in tunnel crossing fibers and at bases of outer hair cells corresponding to the medial system of efferents. The staining seen with antiserum to dynorphin B and to alpha-neoendorphin has similar distribution to that seen with antisera to methionine enkephalin; there may be co-localization of these neuropeptides in the lateral system of efferents. Choline acetyltransferase-like immunoreactivity (co-localized with enkephalin-like immunoreactivity in the lateral system in the brainstem) and glutamic acid decarboxylase (GAD)-like immunoreactivity have also been found in olivocochlear efferents. Further studies will be necessary to determine if the dynorphins are co-localized with other neurotransmitter candidates and what their interactions may be.

Animals↗
PubMed1985

Proenkephalin and prodynorphin related neuropeptides in the cochlea.

Dynorphin B (rimorphin), a proenkephalin B (prodynorphin)-derived peptide, and met-enkephalin-Arg6, Gly7, Leu8 (met-enkephalin octapeptide), a proenkephalin A-derived peptide, were identified in the mammalian cochlea by specific radioimmunoassays. The antisera are directed against unique sequences in the peptides, and this immunoreactivity cannot be ascribed to cross-reaction with any other known opioid peptides. Met-enkephalin octapeptide and dynorphin B can for this reason serve as reliable markers for the proenkephalin A- and proenkephalin B-derived peptides, respectively. Lesion studies in the cochlea indicate that dynorphin B is confined to olivocochlear efferents. It has not been determined if the dynorphin-containing neurons are the same as those known to contain enkephalin-related peptides, or if they may be cholinergic. Different, presumably inhibitory, neurotransmitters or modulators in the olivocochlear fibers create the possibility of separately modulating the effects of inner or outer hair cells on auditory nerve activity, and so becoming able to study their individual actions in audition. The olivocochlear fiber-hair cell-eighth nerve interaction may provide a valuable model for a complex multi-transmitter synaptic junction.

Animals↗
PubMed1984

Several distinct receptor binding enkephalins in olivocochlear fibers and terminals in the organ of Corti.

Biochemical studies centering on the use of reverse-phase high-performance liquid chromatography (HPLC) and radioimmunoassays (RIA) demonstrate the presence in the guinea pig organ of Corti of at least 3 enkephalin-related peptides, two of which are identified as Met- and Leu-enkephalin, respectively. Enkephalins were identified and quantitated by HPLC-RIA in the isolated second turn of the organ of Corti, but were not found in stria vascularis or auditory nerve dissected from the cochlea. Three enkephalin-immunoreactive HPLC fractions inhibited the binding of labeled naloxone to rat brain membranes. All enkephalins identified by the combined HPLC-RIA procedure had an apparent molecular weight similar to that of Met- and leu-enkephalin peptide standards. Immunocytochemistry, performed with the best-characterized Met-enkephalin antiserum used in the RIAs, localized the enkephalin-like immunoreactivity to lateral efferent fibers and terminals under inner hair cells of the organ of Corti. Other antisera raised against Met-enkephalin, not used for RIA, visualized enkephalin-like immunoreactivity in medial efferent fibers under outer hair cells as well. This enkephalin-like immunoreactivity may reflect the presence in the medial efferent system of other structurally similar peptides in addition to those detected biochemically. Efferent fiber lesion, by evulsion of the vestibular nerve close to the vestibulocochlear anastomosis in which the olivocochlear fibers run, eliminated enkephalin-like immunoreactivity and the enkephalin-related peptides identified by HPLC-RIA.

Animals↗
PubMed1984

Immunocytochemical localization of glutaminase-like immunoreactivity in the auditory nerve.

The immunocytochemical localization of glutaminase, which we have proposed as a marker for excitatory amino acid neurotransmitters was determined in the guinea pig auditory nerve. Glutaminase-like immunoreactivity was seen in auditory nerve terminals in the cochlear nucleus and in the cell bodies of the auditory nerve in the cochlea. This staining was seen in type I and not type II spiral ganglion cells. Glutaminase-like immunoreactivity was also observed in granule cells in the cochlear nucleus.

Animals↗
PubMed1984

Enkephalin-like immunoreactivity in the guinea pig organ of Corti: ultrastructural and lesion studies.

Enkephalin-like immunoreactivity (ELI) was examined in a light and electron microscopic study of the normal guinea pig cochlea and of cochlea de-efferented through evulsion of the vestibular nerve. Antiserum to methionine enkephalin, 164, which gives immunoreactive labeling of only the lateral system of efferents, and antiserum 163, which gives immunoreactive labeling of lateral and medial efferents, were used. In de-efferented cochleae no immunoreactive labeling was seen with either antiserum, confirming that in the organ of Corti ELI is confined to efferents. At the ultrastructural level antiserum 163 but not 164 showed ELI in efferent terminals at the base of outer hair cells. ELI with 164 was seen in efferents ending on outer hair cells at the level of the nucleus. Medially, ELI was seen with both antisera in the inner and tunnel spiral bundles. Efferent terminals containing ELI were seen apposing afferent dendrites, other efferents and the inner hair cell. However, only rarely could synaptic contacts be unambiguously identified and then only with afferent dendrites.

Acetylcholinesterase↗
PubMed1984

Glutamic acid decarboxylase immunoreactivity of olivocochlear neurons in the organ of Corti of guinea pig and rat.

The distribution of glutamic acid decarboxylase (GAD)-like immunoreactivity in the organ of Corti of guinea pig and rat was studied under the light microscope. Indirect immunohistochemical techniques were used. Cochleae were first incubated with a specific antiserum to rat brain GAD and then stained through an avidin-biotin-horseradish peroxidase (HRP) procedure. GAD-like immunoreactivity was visualized as staining with HRP reaction product. Surface preparations were prepared from the immunoreacted cochleae. GAD-like immunoreactivity was found in the inner spiral bundle, tunnel spiral bundle, tunnel crossing fibers, outer hair cell synaptic regions and outer spiral bundles. Little staining was seen in the basal turn. Most of the immunoreactivity was seen in the third and lower fourth turn of the guinea pig cochlea, but even there many efferent fibers and endings were unstained. It is concluded that GAD-like immunoreactivity is present in a subpopulation of cochlear efferents that contains elements from both the medial and the lateral olivocochlear system. Future studies are needed to determine whether this subpopulation is GABA-ergic (i.e. uses gamma-aminobutyric acid as a neurotransmitter) and/or cholinergic.

Animals↗
PubMed1984

Colocalization of enkephalin-like and choline acetyltransferase-like immunoreactivities in olivocochlear neurons of the guinea pig.

The guinea pig lateral superior olive was examined immunocytochemically using antisera against enkephalin and choline acetyltransferase sequentially on the same sections. A colocalization of choline acetyltransferase-like and enkephalin-like immunoreactivities was found in cells of the lateral superior olive that give rise to the lateral system of olivocochlear efferents. Only choline acetyltransferase-like immunoreactivity was observed in the group of olivary nuclei that give rise to the medial group of olivocochlear fibers.

Animals↗
PubMed1983

Localization of enkephalin-like immunoreactivity in acetylcholinesterase-positive cells in the guinea-pig lateral superior olivary complex that project to the cochlea.

Olivocochlear fibers have been demonstrated to have acetylcholinesterase-positive staining both in brainstem and cochlea. Olivocochlear fibres in the cochlea have also been determined to contain enkephalin-like immunoreactivity. In this study, we first determined the source of olivocochlear fibers in the guinea-pig using horseradish peroxidase and wheat germ agglutinin in retrograde transport studies. These cells were then examined for enkephalin-like immunoreactivity followed by acetylcholinesterase staining on the same sections to determine which cells and fibers showed staining for both. It was found that cells in the guinea-pig lateral superior olive that project to the cochlea have both enkephalin-like immunoreactivity staining and acetylcholinesterase-positive staining. Cells in other areas giving rise to olivocochlear fibers showed only acetylcholinesterase staining. These results suggest that there is co-localization of enkephalin and acetylcholine in a population of olivocochlear cells and fibers.

Acetylcholinesterase↗
PubMed1983

High-performance liquid chromatographic identification of enkephalin-like peptides in the cochlea.

Met-enkephalin was identified in cochleae from guinea pigs by a combined reversed-phase high-performance liquid chromatographic separation and subsequent radioimmunoassay of the chromatographic fractions. A second Met-enkephalin-immunoreactive fraction was found in the cochlea, with a retention time shorter than that of Met-enkephalin. These findings confirm and extend earlier cytochemical observations, and suggest the differential immunocytochemical staining seen under inner and outer hair cells in the cochlea may be related to different peptides in the two efferent systems innervating the cochlea. Different molecular forms of enkephalin-related peptides may serve different functional or metabolic roles in complex efferent or local regulation of peripheral sensory processing.

Animals↗
PubMed1982

Bicuculline, strychnine and depressant amino acid responses in the anteroventral cochlear nucleus of the cat.

Experiments were conducted in the cat anteroventral cochlear nucleus, comparing the actions of strychnine and bicuculline on amino acid-induced depression of spontaneous and evoked firing. Strychnine reduced responses induced by glycine, taurine and beta-alanine more than GABA or muscimol-induced responses. These latter responses were sensitive to bicuculline. Responses to single and paired tone bursts were not sensitive to strychnine or bicuculline applied either iontophoretically or intravenously. The results indicate that the receptors for depressant amino acids are similar to the receptors found in the cat in other brainstem and spinal cord sites, but differ from those found in the cerebellum, thalamus and cerebral cortex. The data also indicate that these amino acids are not involved either in the response to tone bursts at characteristic frequency or in the suppression of this response by a second, higher frequency tone.

Amino Acids↗
PubMed1982

Aspartate aminotransferase immunoreactivity in cochlea of guinea pig.

The distribution of aspartate aminotransferase-like immunoreactivity in the cochlea of the guinea pig was studied at the light microscopy level. Indirect immunofluorescence histochemistry using antisera against cytoplasmic aspartate aminotransferase prepared from pig heart was applied to surface preparations of the organ of Corti and cryostat sections of the cochlea. In the modiolus, immunofluorescence was localized to spiral ganglion cells and myelinated fibers of the auditory nerve and intraganglionic spiral bundles. In the organ of Corti, immunofluorescence was seen in upper tunnel crossing fibers and at the base of outer hair cells, following a distribution similar to that of the efferent innervation of the outer hair cells. Weak immunofluorescence was seen in the inner spiral bundle and tunnel spiral bundle, but was not present in all preparations. Immunofluorescence was not seen in inner hair cells, nor at the base of inner hair cells, and may have been absent from outer hair cells. It is concluded that spiral ganglion cells and myelinated auditory nerve axons contain aspartate aminotransferase-like immunoreactivity such immunoreactivity has previously been determined in auditory nerve endings inthe cochlear nucleus. Olivocochlear neurons that innervate outer hair cells also contain such immunoreactivity while other cochlear efferents contain little or none.

Animals↗
PubMed1981

Immunocytochemical localization of aspartate aminotransferase immunoreactivity in cochlear nucleus of the guinea pig.

There is substantial evidence supporting the role of aspartate or glutamate as the neurotransmitter of the auditory nerve. The concentration of aspartate aminotransferase (L-aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1), an enzyme associated with the metabolism of these amino acids, is high in axons and terminals of the auditory nerve. Antibodies were raised against aspartate aminotransferase and used in immunocytochemical studies to determine its localization in the cochlear nucleus of the guinea pig. Indirect immunofluorescence techniques were used for light microscopic localization of aspartate aminotransferase-like immunoreactivity in normal guinea pigs and guinea pigs with auditory nerve lesions. Fluorescent rings of aspartate aminotransferase-like immunoreactivity were seen around spherical cells in the anteroventral cochlear nucleus. In animals with auditory nerve lesions, rings were no longer seen in the ipsilateral cochlear nucleus. Immunoreactivity was also seen on cells in the posteroventral cochlear nucleus and in auditory nerve fibers. Ultrastructural studies were done in the rostral anteroventral cochlear nucleus, using the peroxidase-antiperoxidase technique. Aspartate aminotransferase-like immunoreactivity was seen at axosomatic synapses on large spherical cells in terminals with the morphological characteristics of auditory nerve terminals. Other classes of terminals on the soma of large spherical cells showed no immunoreactivity. It was concluded that aspartate aminotransferase-like immunoreactivity is present in axons and terminals of the auditory nerve. These findings indicate that aspartate aminotransferase-like immunoreactivity may serve as a marker at terminals where aspartate or glutamate is a neurotransmitter.

Animals↗
PubMed1981

Enkephalin-like immunoreactivity of olivocochlear nerve fibers in cochlea of guinea pig and cat.

The distribution of enkephalin-like immunoreactivity in the cochlea of the guinea pig and cat was studied. Indirect immunofluorescence immunohistochemistry using antisera generated against a methionine enkephalin-bovine thyroglobulin conjugate was applied to surface preparations of the organ of Corti and cryostat sections of the whole of the cochlea. In the cochlear osseous spiral lamina, immunofluorescence was localized to unmyelinated fibers of the intraganglionic spiral bundle. In the organ of Corti, immunofluorescence was localized to a small number of fibers at inner hair cells, the inner spiral bundle, and tunnel spiral bundle, to tunnel crossing fibers at the level of the tunnel floor, to an occasional spiral outer fiber, and to the synaptic region of outer hair cells in the three rows of the basal turn of the cochlea. Less immunofluorescence was found in this region as one progressed towards the apex, with none seen at the apex. At the most apical region the inner spiral bundle became patchy and the tunnel spiral bundle developed arcades. There was no immunofluorescence found in spiral ganglion cells, in auditory nerve fibers, or in the hair cells of the organ of Corti. The findings were the same in cat as in guinea pig, the latter being studied in more detail. It was concluded that efferent, olivocochlear neurons of the cochlea, synapsing predominantly with primary auditory nerve fibers from the inner sensory cells or with the sensory cells, contain enkephalin-like immunoreactivity. Also, the findings indicate that endings of olivocochlear neurons that synapse predominantly with outer hair cells contain enkephalin-like immunoreactivity. It has previously been shown that olivocochlear neurons are likely to be cholinergic.

Animals↗