Biomedical subjects

E Mugnaini

Publications and source records attributed to E Mugnaini.

At least 73 records · Page 4Linked to original sources

PubMed1991

Chemically distinct rat olivocochlear neurons.

We have produced a neurochemical map of the cell bodies of origin of the cochlear efferent terminals in rat by combining glutamic acid decarboxylase (GAD), choline acetyltransferase (ChAT), or calcitonin gene-related peptide (CGRP) immunocytochemistry with retrograde transport of horseradish peroxidase. The locations of cochlear efferent cell bodies are in general agreement with the medial and lateral systems described by White and Warr (J. Comp. Neurol. 219:203-214, 1983) with some minor modifications. The lateral system consists of at least two pools of chemically distinct neurons located within the lateral superior olive (LSO) ipsilateral to the injected cochlea. One pool immunostains with an antibody to GAD while the other immunostains with antibodies to ChAT and to CGRP. The medial efferent system consists of periolivary neurons that are almost exclusively large and ChAT-positive but CGRP-negative. They are located both ipsilateral and contralateral to the cochlea they project to. There are a few GAD-positive small neurons in the medioventral and rostral periolivary regions that project ipsilaterally, but these may prove tobe ectopic neurons. The ipsilateral lateroventral periolivary region (LVPO) contains some efferent neurons, all of which are ChAT-positive but CGRP-negative. Additional cochlear efferent neurons, some of which are ChAT-positive and others GAD-positive, are present within and immediately dorsal to the fiber capsule surrounding the medial limb, and to a lesser extent the lateral limb, of the ipsilateral LSO. Not all GAD-positive or ChAT-positive olivary cells project to the cochlea. We have complemented the results in the brainstem by demonstrating two immunocytochemically distinct populations of efferent terminals in the cochlea simultaneously, one CGRP-positive and the other GAD-positive. Approximately equal numbers of boutons immunoreactive for both markers are present beneath inner hair cells throughout the entire length of the cochlea. Surprisingly high numbers of GAD-positive and CGRP-positive boutons are also present on outer hair cells, with each class having its spatially and morphologically distinct features. The lack of CGRP-positive periolivary cells that are retrogradely labeled by cochlear injections of HRP suggests that the lateral olivocochlear system sends projections to outer hair cells. Our results raise questions about species differences in the organization of targets of the lateral and medial olivocochlear systems.

Animals↗

PubMed1991

Distribution and targets of the cartwheel cell axon in the dorsal cochlear nucleus of the guinea pig.

This investigation attempted to determine the mode of distribution and synaptic targets of the cartwheel cell axon in the guinea pig dorsal cochlear nucleus (DCoN). Antiserum against PEP-19, a putative calcium-binding neuropeptide, was employed at the light and electron microscopic levels. We show that in the hind-brain of the guinea pig, cerebellar Purkinje cells and DCoN cartwheel cells are the most densely immunoreactive neurons. The PEP-19 immunoreaction product is localized to all neuronal compartments of these cells. Primary targets of cartwheel cell axons are the DCoN pyramidal cells, the large efferent neurons of layer 2. These neurons receive numerous immunoreactive synaptic boutons on their cell bodies and apical and basal dendritic arbors. A PEP-19-immunoreactive axonal plexus, largely formed by cartwheel cell axons, highlights layer 3, co-extensively with the basal arbors of pyramidal cells. This plexus is oriented predominantly in the transstrial plane of the DCoN, in parallel with the sheet-like basal dendritic arbor of pyramidal neurons and with the isofrequency bands of primary cochlear nerve fibers. PEP-19-positive boutons contain pleomorphic synaptic vesicles and form symmetric synaptic junctions, indicative of inhibitory innervation. In addition, immunoreactive boutons, similar to those synapsing on pyramidal neurons, were observed on the cell bodies and main dendritic trunks of cartwheel neurons, indicating a system of recurrent collaterals. Furthermore, a small number of PEP-19-positive axons of unknown origin reach the caudal rim of the posteroventral cochlear nucleus. Within the territory of distribution of the cartwheel cell axon are the dendrites of at least two other types of DCoN neuron, the vertical cells of Lorente de Nó and the giant cells. These neurons may represent additional targets of the cartwheel cell axon, but this remains to be ascertained with specific methods. Our data demonstrate that the cartwheel neurons modulate the activity of pyramidal neurons and, therefore, play a key role in shaping the output of the DCoN superficial layers.

Animals↗

PubMed1991

The GABAergic cerebello-olivary projection in the rat.

Immunocytochemical detection of glutamate decarboxylase (GAD), the predominant biosynthetic enzyme of gamma-aminobutyric acid (GABA), reveals the presence of a dense GABAergic innervation in all parts of the inferior olive. One brain center that provides a substantial projection to the inferior olive is the cerebellar nuclei, which contain many small GABAergic neurons. These neurons were tested as a source of GABAergic olivary afferents by combining retrograde tract tracing with GAD immunocytochemistry. As expected from previous studies, injections of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) into the inferior olive retrogradely label many small neurons in the interposed and lateral cerebellar nuclei and the dorsal part of the lateral vestibular nucleus, and fewer neurons in the ventro-lateral region of the medial cerebellar nucleus. These projections are predominantly crossed and are topographically arranged. The vast majority, if not all, of these projection neurons are also GAD-positive. The relative contribution of this projection to the GABAergic innervation of the inferior olive was tested by lesion of the cerebellar nuclei, or the superior cerebellar peduncle. Within 10 days the lesion eliminates most GAD-immunoreactive boutons in the principal olive, the rostral lamella of the medial accessory olive, the ventrolateral outgrowth, and the lateral part of the dorsal accessory olive ventral fold. Thus, the effectiveness of this depletion demonstrates that the cerebellar nuclei provide most of the GABAergic innervation to regions of the inferior olive known to receive a cerebellar projection. Moreover, when the lateral vestibular nucleus is damaged, the dorsal fold of the dorsal accessory olive is depleted of GABAergic boutons. The synaptic relations that boutons of the GABAergic cerebello-olivary projection share with olivary neurons were investigated at the electron microscopic level by GAD-immunocytochemistry, anterograde degeneration of the cerebellar axons or anterograde transport of WGA-HRP. All of these methods confirm that GABAergic, cerebello-olivary axon terminals contain pleomorphic vesicles, and synapse on various portions of olivary neurons, and especially on dendritic spines within glomeruli, often in very close proximity to the gap junctions that characteristically couple the dendritic profiles. These results demonstrate four major points: that virtually all of the GABAergic, and presumably inhibitory, neurons of the cerebellar and dorsal lateral vestibular nuclei are projection neurons; that a large portion of the inferior olive receives GABAergic afferents from the cerebellar nuclei; that a portion of the dorsal accessory olive receives GABAergic afferents from the dorsal lateral vestibular nucleus; and that cerebello-olivary fibers often synapse near gap junctions, and therefore could influence electrical coupling of olivary neurons.

Animals↗

PubMed1990

Stiff-man syndrome: a GABAergic autoimmune disorder with autoantigenic heterogeneity.

Autoantibodies that reacted with cell bodies and axon terminals of gamma-aminobutyric acid (GABA)ergic neurons were present in the serum and cerebrospinal fluid in a patient with stiff-man syndrome with type I diabetes. Immunoblot experiments using this patient's serum and cerebrospinal fluid did not corroborate an earlier observation that these autoantibodies are directed against the GABAergic cytosolic enzyme, L-glutamic acid decarboxylase. While L-glutamic acid decarboxylase autoantibodies may be associated with this syndrome, they do not appear to be easily demonstrated.

Adult↗

PubMed1990

The Purkinje cell class may extend beyond the cerebellum.

The cerebellum develops from a germinal zone at the rhombic lip of the metencephalon. This region, like the telencephalic vesicle which gives rise to the cerebral cortex, presumably consists of germinative units showing a rather repetitive neurogenetic pattern. In all cerebellar folia, cortical neurons, and especially the Purkinje cells, express highly stereotyped phenotypes, although some variations in their chemical make-up have been uncovered with monoclonal antibodies. Here, we demonstrate for the first time that three independent murine mutations, Lurcher, Purkinje cell degeneration and staggerer, which result in the postnatal degeneration of Purkinje cells, also cause the elimination of cartwheel cells of the dorsal cochlear nucleus. The cerebellar granule cell mutation, weaver, which spares most Purkinje cells in the lateral cerebellum, also spares cartwheel cells. These data support the notion that the cerebellar germinative zone extends to the caudal portion of the rhombic lip, which gives rise to the dorsal cochlear nucleus.

Animals↗

PubMed1990

Immunocytochemical evidence for inhibitory and disinhibitory circuits in the superior olive.

Immunostaining of the cat superior olivary complex with antisera against glycine, calbindin, GABA, and its synthetic enzyme, glutamate decarboxylase (GAD) reveals considerable new information about neural connections that underlie processing of binaural signals. Antisera against glycine and calbindin immunostain principal cells and processes of the medial nucleus of the trapezoid body (MNTB). The extent of staining of fine processes afforded by the calbindin antisera reveals collateral processes of MNTB axons within the medial superior olive (MSO) and numerous terminals of these collaterals on neuronal somata and proximal dendrites. Electron microscopic immunocytochemistry of these terminals shows them to have the morphological features characteristic of inhibitory synapses, indicating that there can be considerable inhibitory inputs to MSO cells from the contralateral cochlear nucleus via the MNTB. Immunostaining for GAD and GABA shows some GABAergic inputs, mostly to dendrites within the MSO. Within the MNTB there are elaborate GABAergic endings that surround the principal cells. Electron microscopic immunocytochemistry of these terminals reveals the expected features characteristic of inhibitory synapses. The glycinergic inhibitory output of the MNTB cells is therefore subject to disinhibition by activity of these elaborate GABAergic inputs. Other endings that immunostain with antisera to leucine- enkephalin and neurotensin are found in modest amounts in both MSO and MNTB. Immunostaining of neuronal somata within the superior olivary complex suggests that the origins of the peptidergic and GABAergic endings in the MSO and MNTB may be periolivary cells. Whatever the origins, the results clearly indicate that control of the inhibitory GABAergic inputs to the MNTB can profoundly affect the inhibitory glycinergic MNTB outputs to other principal olivary nuclei. Investigations of the circumstances under which these circuits are activated will reveal much about neural processing that underlies binaural hearing.

Animals↗

PubMed1990

An evaluation of retrograde tracing methods for the identification of chemically distinct cochlear efferent neurons.

We have compared retrograde labelling of rat olivocochlear neurons after unilateral cochlear injections of wheatgerm agglutinin conjugated horseradish peroxidase (WGA-HRP) and free HRP. After cochlear injection of WGA-HRP, labelling of nerve cell bodies in the brainstem can be explained not only as conventional retrograde labelling resulting from uptake by efferent nerve terminals synapsing on or near hair cells, but also as spurious labelling originating from tracer leakage, through the periotic duct and over the eighth nerve sheaths, into the cerebral-spinal fluid. Depending on the length of survival time, spurious labelling can involve small portions of the nucleus of the trapezoid body or the entire auditory brainstem and other non-auditory centers. On the contrary, moderate amounts of free HRP delivered to the cochlea do not lead to spurious labelling. With free HRP as the tracer of choice, we found that cochlear efferent cells were located not only in the ipsilateral LSO body and bilaterally within MVPO and RPO as already described by White and Warr, but also surrounding the ipsilateral LSO and in the ipsilateral LVPO. The allocation of these newly described olivocochlear neurons to the medial large cell or lateral small cell system is uncertain because they are located laterally in the brainstem and project ipsilaterally but are large spherical to fusiform or multipolar cells. A zinc salicylate-formol fixative and a metal intensified DAB reaction were found to be effective in visualizing retrogradely transported HRP in neurons and allowed immunocytochemical staining of the same sections with antisera to glutamic acid decarboxylase and choline acetyltransferase. This double label protocol can be used to produce a neurochemical map of the OC systems.

Animals↗

PubMed1989

Comparative study of glutamate decarboxylase immunoreactive boutons in the mammalian inferior olive.

An antiserum raised against rat glutamate decarboxylase was used to map GABAergic boutons in the inferior olive of rabbit, cat, rhesus monkey, and human. A description of the human periolivary region is also included. The inferior olive of each species contained a dense GABAergic innervation, but immunostaining intensities varied among regions. These intensities were evaluated visually and photometrically, and the sizes and frequencies of occurrence of boutons in various olivary subnuclei were measured. The beta nucleus in all species was intensely immunostained and contained the largest boutons. The caudal subdivision of the dorsal accessory olive stained with a lower intensity than the beta nucleus, but contained similarly large GABAergic boutons. By visual analysis, the rostral subdivision and the subnucleus a of the medial accessory olive and the principal olive were stained with an intermediate intensity, and these regions contained small GABAergic boutons. Photometric analysis of focal regions of the neuropil, however, revealed species differences in teh staining intensity of the principal olive, which was lowest in rabbits and highest in primates. In all species, the lowest immunostaining intensity was observed in the subnucleus b of the medial accessory olive. Species variations in bouton sizes and regional staining intensities were observed in the dorsal cap and the dorsomedial cell column. The heterogeneous staining pattern and regional variation of bouton size argue for the existence of separate GABAergic projections to discrete regions of the inferior olive. Since glutamate decarboxylase immunostaining patterns in the olive are largely similar across species, the afferent projections producing these patterns may also be similar.

Adolescent↗

PubMed1988

Cerebellin and related postsynaptic peptides in the brain of normal and neurodevelopmentally mutant vertebrates.

The rat cerebellum was previously shown to contain two polypeptides, a hexadecapeptide termed cerebellin and an apparent metabolite des-Ser1-cerebellin. The cerebellins have a high degree of sequence homology with residues 625-641 of the polyimmunoglobulin (polyIg)-receptor adjacent to its membrane-spanning domain. Since the cerebellins are localized in Purkinje cells and enriched in synaptosomes, this might indicate that cerebellin is a specific proteolytic cleavage fragment of a synaptic protein involved in the transcytosis of an unknown ligand. Using a specific cerebellin radioimmunoassay described here combined with high-performance liquid chromatography, cerebellin immunoreactivity could be demonstrated in the cerebella of all vertebrates examined from man to chicken. Cerebellin immunoreactivity is localized to Purkinje cells in the rat, mouse, and chicken. Furthermore, cerebellin expression is under developmental regulation in both the chicken and mouse. In addition, neurodevelopmental mutations of mice that eliminate granule cells cause a large deficit in cerebellin levels, suggesting some form of transneuronal regulation.

Amino Acid Sequence↗

PubMed1988

Cerebellin is a postsynaptic neuropeptide.

Cerebellin is a hexadecapeptide that has been biochemically characterized and localized to cerebellar Purkinje cells and certain neurons of the dorsal cochlear nucleus (DCoN) of rat. Among rabbit antisera produced to synthetic cerebellins, one (C1) gave specific immunostaining of the Purkinje neuronal cell body, initial axon segment, and main stem dendrites, while another (R2) reacted with peripheral dendritic structures. This complementarity of staining was also present during cerebellar development. By electron microscopy, the immunoreaction product was localized to polyribosomal domains with antiserum C1 and to dendritic spines with antiserum R2, in both cerebellar cortex and DCoN. In the spine, the structure most strongly stained was the postsynaptic density, but some reaction product was adsorbed to the plasma membrane, the spine apparatus, and the granulofibrillar cytoplasmic component. Antiserum R2 also stained lysosome-like bodies. We suggest that antiserum C1 recognizes cerebellin precursor(s) and antiserum R2 mature peptide(s) and perhaps degradation product. There is structural homology between cerebellin and residues 625-641 of the polyimmunoglobulin transporter. The functional implications of this homology and other possible roles of cerebellin are discussed.

Animals↗

PubMed1988

The rat inferior olive as seen with immunostaining for glutamate decarboxylase.

Boutons presumed to use gamma-aminobutyric acid as neurotransmitter (GABAergic boutons) were detected by glutamate decarboxylase (GAD) immunocytochemistry in all regions of the rat inferior olive. The remarkably high concentration of these boutons allowed a clear visualization of olivary subnuclei boundaries. Regional variations in GAD immunostaining intensity were observed within the nuclear complex and were graded both visually and photometrically. The regional staining variations, for the most part, followed subnuclei boundaries and olivary zonal compartments that have been delineated by the topography of climbing fibre projections. Some subnuclei were grouped by similar staining intensities. The beta nucleus and a medial region in the ventral fold of the dorsal accessory olive were most intensely immunostained, followed by the subnucleus c of the medial accessory olive. Lower staining intensities were observed in the dorsomedial cell column, the dorsal fold of the dorsal accessory olive and the dorsal cap. The lowest intensities were observed in the subnuclei a and b of the medial accessory olive, the ventrolateral outgrowth, the rostral lamella of the medial accessory olive, the principal olive, and the lateral part of the ventral fold of the dorsal accessory olive. The factors contributing to the variations in immunostaining intensity (bouton size and frequency of occurrence) were investigated. The largest boutons were observed in the beta nucleus. Intermediate sized boutons were observed in the dorsomedial cell column, dorsal cap and the dorsal fold of the dorsal accessory olive. The smallest boutons were present in the remaining regions of the inferior olive, including the principal olive, the rostral lamella of the medial accessory olive, and the ventral fold of the dorsal accessory olive. The medial region of the dorsal accessory olive ventral fold contained a higher density of GABAergic boutons than other regions. GABAergic bouton size and innervation density therefore largely accounted for the variations in GAD immunostaining intensity. This study provides a map of the rat inferior olive based on the distribution of GABAergic nerve terminals, and may serve as a basis for characterizing different GABAergic afferent systems in the inferior olive.

Animals↗

PubMed1988

Effects of the murine mutation 'nervous' on neurons in cerebellum and dorsal cochlear nucleus.

'Nervous' mutant mice are presently available on two different genetic background strains which are derived from out-breeding of the original BALB/cGr mutant stock. Light and electron microscopic studies of these mutants demonstrate that cerebellar Purkinje cells and cartwheel neurons of the dorsal cochlear nucleus (DCoN) show similar, albeit not identical, cytoplasmic and mitochondrial alterations in both background strains. In the cerebellar cortex, all Purkinje cell perikarya developed a varying number of enlarged and rounded mitochondria, as previously described. Extensive changes were observed in various components of the mitochondrial matrix. As cellular degeneration proceeded, reduction, fragmentation and dilation of cisterns of endoplasmic reticulum and the Golgi apparatus were evident. Some of the mitochondria underwent a peculiar type of degeneration, i.e. the outer membrane partially or completely dissolved, occasionally accompanied by focal interruptions of the inner membrane. In older adult mutants only 10% of cerebellar Purkinje cells rehained. The few surviving cells displayed varying states, ranging from essentially normal ultrastructure to electron-dense condensation. Many of these cells, in both strains, continued to display greatly enlarged, rounded mitochondrial profiles, indicating a change in the expression of the gene defect resulting from genetic contamination. Criteria for the identification of neuronal cell classes in layers 1 and 2 of murine DCoN were established. Cartwheel neurons in the mutant DCoN presented alterations similar to those observed in cerebellar Purkinje cells. The characteristic mitochondrial anomaly developed and proceeded in cartwheel neurons within a comparable time frame. The vast majority of affected cartwheel cells did not undergo degeneration, however, but continued to possess altered mitochondria into adulthood. The differences between normal and mutant mitochondria in Purkinje and cartwheel were quantified by morphometric analyses. Our findings lend support to the notion of a homology between cerebellar Purkinje cells and DCoN cartwheel cells. These cells represent major elements in two similar spatially related circuits, and share several genetic, structural and neurochemical properties. It is therefore proposed that these two cell populations are derived from closely related precursor cells.

Aging↗

PubMed1987

Patterns of glutamate decarboxylase immunostaining in the feline cochlear nuclear complex studied with silver enhancement and electron microscopy.

The cochlear nuclear complex of the cat was immunostained with an antiserum to glutamate decarboxylase (GAD), the biosynthetic enzyme for the inhibitory neurotransmitter GABA, and studied with different procedures, including silver intensification, topical colchicine injections, semithin sections, and immunoelectron microscopy. Immunostaining was found in all portions of the nucleus. Relatively few immunostained cell bodies were observed: most of these were in the dorsal cochlear nucleus and included stellate cells, cartwheel cells, Golgi cells, and unidentified cells in the deep layers. An accumulation of immunoreactive cells was also found within the small cell cap and along the medial border of the ventral cochlear nucleus. Immunostained cells were sparse in magnocellular portions of the ventral nucleus. Most staining within the nucleus was of nerve terminals. These included small boutons that were prominent in the neuropil of the dorsal cochlear nucleus, the granule cell domain, in a region beneath the superficial granule cell layer within the small cell cap region, and along the medial border of the ventral nucleus. Octopus cells showed small, GAD-positive terminals distributed at moderate density on both cell bodies and dendrites. Larger, more distinctive terminals were identified on the large cells in the ventral nucleus, in particular on spherical cells and globular cells. There was a striking positive correlation of the size, location, and complexity of GAD-positive terminals with the size, location, and complexity of primary fiber endings on the same cells. This correlation did not hold in the dorsal nucleus, where pyramidal cells receive many large GAD-positive somatic terminals despite the paucity of primary endings on their cell bodies. The GAD-positive terminals contained pleomorphic synaptic vesicles and formed symmetric synaptic junctions that occupied a substantial portion of the appositional surface to cell bodies, dendrites, axon hillocks, and the beginning portion of the initial axon segments. Thus, the cells provided with large terminals can be subjected to considerable inhibition that may be activated indirectly through primary fibers and interneurons or by descending inputs from the auditory brainstem.

Animals↗

PubMed1987

Cytology and immunocytochemistry of the nucleus of the lateral line lobe in the electric fish Gnathonemus petersii (Mormyridae): evidence suggesting that GABAergic synapses mediate an inhibitory corollary discharge.

Knollenorgans, low-threshold electroreceptors found in mormyrid fish, are involved primarily, if not exclusively, in communication. The rhombencephalic nucleus of the lateral line lobe (nLLL) is the target nucleus of knollenorgan afferents. Cells in the nLLL receive a few medium size to large endings with round synaptic vesicles (classified as spoon; large club; small club-, and rodlet-shaped endings) with which they form nexus (gap junction) and asymmetrical chemical synapses associated with the round synaptic vesicles. In addition these endings emit thin collaterals which terminate as small boutons on nLLL neurons; these boutons also have round vesicles and make mixed (electrotonic and chemical) synapses. In addition, cells in the nLLL receive synaptic input from numerous small boutons containing pleomorphic vesicles and making symmetric synapses. We have not found any interneurons within nLLL. Our ultrastructural analysis suggests that boutons synapsing on nLLL neurons belong to only two afferent fiber systems and that the wiring diagram of nLLL is extremely simple. We have studied the immunolocalization in nLLL of glutamic acid decarboxylase (GAD), the enzyme essential for the synthesis of GABA that is also a useful marker for this widely distributed inhibitory neurotransmitter. GAD immunoreactivity was confined to the small boutons with pleomorphic vesicles. GAD was also found in a nucleus projecting to the nLLL, here named the sublemniscal nucleus (SL), which probably conveys corollary discharge signals to the nLLL. This GABAergic projection may be responsible for the potent inhibition associated with the electric organ discharge command that has been described in these cells.

Animals↗

PubMed1987

Cytology and immunocytochemistry of the nucleus extrolateralis anterior of the mormyrid brain: possible role of GABAergic synapses in temporal analysis.

Mormyrid fish use their electrosensory/electromotor system for both electrolocation and electrocommunication. One type of electroreceptor, the knollenorgan, has a low threshold and is used strictly for detecting a conspecific's electric organ discharge. Knollenorgan afferents terminate with mixed synapses (gap junctions and asymmetric chemical synapses) on neurons of the nucleus of the lateral line lobe (nLLL), which in turn projects to a midbrain nucleus--the nucleus extrolateralis anterior (ELa). ELa contains numerous granule cells, which are adendritic, and scattered larger neurons, here termed interstitial cells, which possess several branched dendrites. Because of the overall paucity of dendrites, the neuropil of ELa is scarcely developed and myelinated fibers predominate in between the cell bodies. Axons presumed to originate in nLLL make mixed synapses on both the interstitial and the granule cells. The interstitial neurons are immunoreactive for glutamic acid decarboxylase (GAD), the synthetic enzyme for the inhibitory neurotransmitter gamma-aminobutyric acid (GABA), while the granule cells are GAD-negative. Many GAD-positive synapses are found on each granule cell soma; at smaller densities, GAD-positive boutons also synapse on the cell bodies and dendrites of interstitial cells. The GAD-positive boutons have pleomorphic vesicles and make symmetric synapses. Such boutons presumably originate from a plexus of interstitial cell axons. Mormyrid fish use their knollenorgan afferents for precise temporal discriminations, and this information is needed for electrocommunication. The ELa, with its relatively simple neuronal circuitry, is the most likely site for precise temporal analysis in the knollenorgan afferent pathway, and we propose that the GABAergic synapses on the granule cells may be essential for accurate detection of specific time intervals.

Animals↗

PubMed1987

The neuropeptide cerebellin is a marker for two similar neuronal circuits in rat brain.

We report here that the neuropeptide cerebellin, a known marker of cerebellar Purkinje cells, has only one substantial extracerebellar location, the dorsal cochlear nucleus (DCoN). By reverse-phase high-performance liquid chromatography and radioimmunoassay, cerebellum and DCoN in rat were found to contain similar concentrations of this hexadecapeptide. Immunocytochemistry with our rabbit antiserum C1, raised against synthetic cerebellin, revealed that cerebellin-like immunoreactivity in the cerebellum is localized exclusively to Purkinje cells, while in the DCoN, it is found primarily in cartwheel cells and in the basal dendrites of pyramidal neurons. Some displaced Purkinje cells were also stained. Although cerebellum and DCoN receive their inputs from different brain regions, their organizations show several similarities, with many neuronal cell types present in one having their presumed homologue in the other. Cerebellin is present in Purkinje and cartwheel cells throughout the cerebellum and the DCoN without site predilection. This should be viewed together with recent data indicating that these two classes of neurons share a similar fate in several murine mutations ("nervous," "lurcher," "Purkinje cell degeneration," and "staggerer"). This situation leads us to conclude that cerebellin represents a marker for a small number of spatially related neuronal cell classes, and that it may play a role in an as yet unknown aspect of a circuit function worthy of further exploration.

Animals↗

PubMed1987

The polypeptide PEP-19 is a marker for Purkinje neurons in cerebellar cortex and cartwheel neurons in the dorsal cochlear nucleus.

This light and electron microscopic immunocytochemical study shows that the polypeptide PEP-19, a presumptive calcium binding protein specific to the nervous system, represents an excellent marker for cerebellar Purkinje cells and dorsal cochlear nucleus (DCoN) cartwheel cells. The polypeptide clearly reveals the entire populations of both types of neurons, including their complete dendritic and axonal arborizations. Other PEP-19 containing neurons in the two regions display weak immunoreactivity restricted to the cell body or to cell body and principal dendrites. Electron microscopic localization of PEP-19-like immunoreactivity reveals similarities between this polypeptide, parvalbumin, and a 28K vitamin D-dependent calcium binding protein. However, calmodulin, which is expressed in both Purkinje and granule cells, may differ from PEP-19. Similarities between the organization of the cerebellar cortex and the DCoN superficial layers have been known for some time, with several types of neurons in one system having their presumed homologue in the other. These data provide further support for the proposed structural and functional homology between Purkinje and cartwheel neurons, and establishes PEP-19 as a useful marker for examining degeneration of these two neuronal populations in murine cerebellar mutants.

Animals↗

PubMed1985

Projection of olfactory bulb efferents to layer I GABAergic neurons in the entorhinal area. Combination of anterograde degeneration and immunoelectron microscopy in rat.

Glutamic acid decarboxylase (GAD) immunoelectron microscopy in combination with anterograde degeneration was applied in rats to study the synaptic targets of olfactory bulb afferents to the lateral subdivision (LEA) of the entorhinal area (EA). Immunoreactive neurons and terminals are scattered throughout all layers of LEA. After olfactory bulb resection, terminal degeneration occurs in layer Ia of EA. Using the electron microscope we examined serial thin sections of 12 and 14 immunoreactive neurons sampled from layer Ia of the dorsal (DLEA) and ventral (VLEA) subdivisions of LEA, respectively. The morphology of all these neurons is similar: they are small (short axis 5-9 micron, long axis 7-12 micron) and possess eccentrically located, indented nuclei provided with filamentous nuclear rodlets. The immunoreactive neurons have thin, smooth dendrites which usually emerge abruptly from the somata. We observed a single cilium on 5 of the immunoreactive neurons. In layer Ia of both DLEA and VLEA, the somata of the immunoreactive neurons are contacted by degenerating, non-immunoreactive boutons showing asymmetric synaptic junctions. In addition to these boutons, 4 other categories of axo-somatic terminals can be distinguished: normal, non-immunoreactive boutons forming asymmetric synapses and containing spherical synaptic vesicles; normal, non-immunoreactive boutons with symmetric synapses and pleomorphic synaptic vesicles; normal, non-immunoreactive boutons with asymmetric synapses, containing dense-cored vesicles in addition to spherical synaptic vesicles; and normal, immunoreactive boutons with symmetric synapses and pleomorphic synaptic vesicles. It is suggested that the GAD-immunoreactive neurons which receive olfactory bulb input correspond to local circuit neurons with intralaminar axons which innervate each other as well as the distal segments of the apical dendrites of projection neurons with cell bodies in layers II and III. Thus, the olfactory input in EA seems to be wired not only for excitation of layers II and III pyramidal neurons but also for feed-forward inhibition using GABAergic intermediary neurons, strategically located in the area of termination of olfactory bulb fibers.

Animals↗