Kirsten K. Osen

Anatomy of the inferior colliculus in rat

SummaryThis paper defines the pattern of subdivision of the inferior colliculus in rat. It is based on serial sections of brains of albino and hooded rats cut in the frontal, sagittal and horizontal planes using Golgi, Nissl and a combined cell-myelin method. In rat, like in other mammals, the inferior colliculus consists of a central nucleus, an external cortex, and a dorsal cortex. The central nucleus is flattened in the frontal plane and confined to the caudomedial part of the inferior colliculus. It is characterized by a lamellar organization of disc-shaped neurons interspersed with multipolar cells. The cells are small to medium-sized. Although there is a dorsoventral gradient in size and packing density of cells within the nucleus, the overall size is smaller and the packing density larger than in adjacent subdivisions. The two cortices each consists of three layers. The outer-most layer is common to the two cortices, forming a fibro-cellular capsule continuous along most of the circumference of the inferior colliculus. The external cortex is located lateral, rostral, ventral and ventrocaudal to the central nucleus. Its second layer, deep to the superficial capsule, is characterized by clusters of many small and a few medium-sized neurons in a myelin-dense neuropil. Layer 3, which constitutes the major portion of the subdivision, consists of relatively scattered, small, medium and large cells, the most characteristic element being large multipolar neurons with coarse Nissl granules. The dorsal cortex is located dorsocaudal and dorsomedial to the central nucleus. Its second layer is composed of small neurons, while the third, deep layer in addition contains medium-sized neurons. The cell density is intermediate to that of the central nucleus and the deep part of the external cortex. We have tried to facilitate the parcellation by reference to easily recognizable, nearby structures and to standard stereotaxic coordinates.

An atlas of glycine- and GABA-like immunoreactivity and colocalization in the cochlear nuclear complex of the guinea pig

SummaryThe distribution and colocalization of γ-aminobutyric acid (GABA)- and glycine-like immunoreactivity in the cochlear nuclear complex of the guinea pig have been studied to produce a light microscopic atlas. The method used was based on post-embedding immunocytochemistry in pairs of 0.5-μm-thick plastic sections treated with polyclonal antibodies against conjugated GABA and glycine respectively. Immunoreactive cells, presumably short axon neurones, predominated in the dorsal cochlear nucleus, with mostly single-GABA-labelled cells in the superficial layer, double-labelled in the middle, and single-glycine-labelled in the deep layers. A few large single-glycine-labelled cells, interpreted as commissural neurons, occurred in the ventral nucleus. Scattered double-labelled cells, probably Golgi cells, were seen in the granule cell domain. Immunolabelled puncta of all three staining categories occurred in large numbers throughout the complex, apposed to somata and in the neuropil, showing a differential distribution onto different types of neuron. Three immunolabelled tracts were noted: the tuberculoventral tract, the commissural acoustic stria, and the trapezoidal descending fibres. Most of the fibres in these tracts were single-labelled for glycine, although in the last mentioned tract single-GABA- and double-labelled fibres were also found. Some of the immunolabelled cell types described here are proposed as the origins of the similarly labelled puncta and fibres on the basis of known intrinsic connections.

Anatomy of the cochlear nuclear complex of guinea pig

SummaryThe cyto-and fibre-architecture of the cochlear nuclear complex of the guinea-pig has been studied in serial sections using Nissl, Golgi and combined cellmyelin staining of normal material, and a silver degeneration method after cochlear ablation. The nuclear subdivisions and major cell types can be recognised on the basis of those found in the cat, but there are some differences between the two species in the precise distribution and morphology of the neurons. The rostrodorsal part of the anteroventral cochlear nucleus (AVCN) contains predominantly spherical bushy cells, but these cannot be readily divided into large and small types as in the cat. Globular bushy cells are seen in the caudal region of the AVCN, but the majority occur in the posteroventral cochlear nucleus (PVCN), in an area extending from the nerve root right up to the boundary of the dorsal cochlear nucleus (DCN). The octopus cells constitute a distinct region in the most dorsomedial part of the PVCN underneath the DCN. Giant cells are seen scattered around the nerve root region. Multipolar and small cells are seen throughout the non-granular regions of the ventral cochlear nucleus (VCN) except for the octopus cell area, but occur mainly in the more rostral regions of the PVCN. Small cells occur in greatest abundance in the thin cap area at the dorsal edge of the VCN below a superficial granule cell layer. The latter covers the dorsolateral surface of the VCN, and a lamina of granule cells partially separates the PVCN from the DCN. The DCN can be divided into four layers. The outermost molecular layer (layer 1) is separated from the deeper regions by a prominent layer of granule cells (layer 2) which also contains the pyramidal cells. Molecular layer stellate cells are seen in layer 1 and a staggered row of cartwheel neurons is found at the boundary between layers 1 and 2. Layer 3 contains the basal dendrites of the pyramidal cells and some small (vertical) cells, and is innervated by the descending branches of the cochlear nerve. The deepest layer 4, which contains multipolar cells and giant cells, does not appear to receive this direct cochlear input.

Histochemical localization of cholinesterases in the cochlear nuclei of the cat, with notes on the origin of acetylcholinesterase-positive afferents and the superior olive.

Abstract This study was carried out in order to correlate the distribution of cholinesterases in the cat cochlear nuclei with the cytoarchitecture and terminal areas of afferent fiber tracts. Formaldehyde-fixed frozen sections were stained by the thiocholine method using acetylthiocholine and butyrylthiocholine iodides as substrates and Mipafox as a specific inhibitor of butyrylcholinesterase (BuChE). The acetylcholinesterase (AChE) activity appeared to be associated with neurons while the BuChE reaction probably was confined to glial elements. Short incubation revealed only a few AChE positive structures which appeared to form a continuous pathway leading from certain cells in the superior olivary complex via the olivocochlear bundle to the molecular and granular cell layers of the cochlear nuclei. The probable role of this AChE-positive pathway in the central control of the pyramidal cell activity in the cochlear nuclei is discussed. Prolonged incubation resulted in a more widespread AChE reaction, and in these sections most cell areas of the cochlear nuclei could be defined by their staining properties. The AChE-positive plexus of the large and small spherical cell areas may represent the terminal arborization of the facilitatory cholinergic pathway found physiologically by Whitfield and Comis50,51 to arise in the lateral superior olive on the same side.

Pyramidal neurones of the dorsal cochlear nucleus: A golgi and computer reconstruction study in cat

The main projection neurones of the dorsal cochlear nucleus, termed pyramidal, bipolar or fusiform cells, have an apical dendritic arbor approaching the ependymal surface of the nucleus and a basal arbor oppositely directed. In Golgi-Del Rio-Hortega material these neurones were studied, with the light microscope, in nonconventional planes of sectioning oriented across or parallel to the main axis of the elongated nucleus. The pyramidal neurones were seen to be flattened across this axis. The size, shape and orientation of 21 cells from six blocks were studied in detail with computer-aided graphic reconstructions including stereo views. Camera lucida drawings of each cell (usually from several sections) were digitized to obtain x and y coordinates while z coordinates (depths in the tissue) were read from the fine focus knob during microscopy and typed interactively during digitization. The z values were corrected for the effects of refractive index differences in the optical system. Since it was the aim of this study to focus on some fundamental principles of structure and arrangement of pyramidal cells in the dorsal cochlear nucleus rather than on topographic variations, only the middle, regularly built part of the nucleus was examined. Towards the ends of the nucleus the architecture is less regular and will require separate analysis. Measurements of arbor and total cell height and of dendritic length are given. The height of the apical and basal arbor in individual cells showed considerable reciprocity. The total dendritic length was up to 8300 micron (average 6536 micron). The basal arbors always proved to be conspicuously flattened; roughly, the width varied between about 300 and 700 micron (average 489 micron) and the thickness between 65 and 105 micron (average 80 micron). The apical arbors were also often flattened but much less and with a greater variability than the basal arbors (average width 319 micron, thickness 115 micron). The two arbors of individual cells were practically coplanar, the arbor planes showing only moderate angularity (bend) and/or torsion relative to each other (angularity maximum 10 degrees, average 5 degrees; torsion maximum 18 degrees, average 6 degrees). The mutual orientation of cells from the same block was examined. The planes through the basal arbors proved to be very parallel, the differences in orientation angles being between 10 and 0 degrees with rare exceptions. Clearly flattened, apical arbors showed a somewhat greater spread.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell-specific expression of the glutamine transporter SN1 suggests differences in dependence on the glutamine cycle.

Glutamine is involved in a variety of metabolic processes, including recycling of the neurotransmitters glutamate and γ‐aminobutyric acid (GABA). The system N transporter SN1 mediates efflux as well as influx of glutamine in glial cells [Chaudhry et al. (1999), Cell, 99, 769–780]. We here report qualitative and quantitative data on SN1 protein expression in rat. The total tissue concentrations of SN1 in brain and in kidney are half and one‐quarter, respectively, of that in liver, but the average concentration of SN1 could be higher in astrocytes than in hepatocytes. Light and electron microscopic immunocytochemistry shows that glutamatergic, GABAergic and, surprisingly, purely glycinergic boutons are ensheathed by astrocytic SN1 laden processes, indicating a role of glutamine in the production of all three rapid transmitters. A dedication of SN1 to neurotransmitter recycling is further supported by the lack of SN1 immunoreactivity in oligodendrocytes (cells rich in glutamine but without perisynaptic processes). All neuronal structures appear unlabelled implying that a different protein mediates glutamine uptake into nerve endings. In several regions, SN1 immunoreactivity is higher in association with GABAergic than glutamatergic synapses, in agreement with observations that exogenous glutamine increases output of transmitter glutamate but not GABA. Nerve terminals with low transmitter reuptake or high prevailing firing frequency are associated with high SN1 immunoreactivity in adjacent glia. Bergmann glia and certain other astroglia contain very low levels of SN1 immunoreactivity compared to most astroglia, including retinal Müller cells, indicating the possible existence of SN isoforms and alternative mechanisms for transmitter recycling.

The Intrinsic Organization of the Cochlear Nuclei in the Cat

In the cochlear nuclei nine different cell types are distinguished on the basis of Nissl preparations. Only three of these cell types are presented here. The large and small spherical cells are innervated from the ascending cochlear branches by means of typical bulbs of Held. The former cell group is apparently supplied only from the apical and middle part of the cochlea and projects bilaterally on the medial superior olive, while the latter group seems to be supplied from the entire cochlea and projects on the lateral superior olive on the same side. The octopus cells are innervated from the descending cochlear branches by means of small ring-shaped boutons. The axons of these cells run in the intermediate acoustic stria and terminate probably in the retroolivary and medial preolivary nuclei on both sides. The function of the three types of cells is discussed.

Colocalization of GABA and glycine in the ventral nucleus of the lateral lemniscus in rat: An in situ hybridization and semiquantitative immunocytochemical study

We have studied by in situ hybridization for GAD65 mRNA in thick sections and by semiquantitative postembedding immunocytochemistry in consecutive semithin sections, the expression of γ‐aminobutyric acid (GABA) and glycine in cell bodies and axosomatic puncta of the rat ventral nucleus of the lateral lemniscus (VNLL), a prominent monaural brainstem auditory structure. The in situ hybridization and the densitometric analysis of the immunostaining suggest that the rat VNLL contains two main populations of neurons. Approximately one‐third of neurons are unstained with either technique and are presumably excitatory; their cell bodies are enveloped by a large number of glycine‐immunoreactive puncta. Most if not all of the remaining two‐thirds colocalize GABA and glycine and are assumed to be inhibitory. These two populations show a complementary distribution within the VNLL, with inhibitory neurons located mainly ventrally and excitatory neurons dorsally. In scatterplots of gray values measured from cell bodies, the double‐labeled cells appear to form a single cluster in terms of their staining intensities for the two transmitter candidates. However, this cluster may have to be further subdivided because cells with extreme GABA/glycine ratios differ from those with average ratios with respect to location or size. The VNLL seems unique among auditory structures by its large number of neurons that colocalize GABA and glycine. Although the functional significance of this colocalization remains unknown, our results suggest that the VNLL exerts convergent excitatory and inhibitory influences over the inferior colliculus, which may underlie the timing processing in the auditory midbrain. J. Comp. Neurol. 432:409–424, 2001.

Expression of glutamine synthetase and glutamate dehydrogenase in the latent phase and chronic phase in the kainate model of temporal lobe epilepsy.

It has been suggested that astrocytic glutamate release or perturbed glutamate metabolism contributes to the proneness to epileptic seizures. Here we investigated whether astrocytic contents of the major glutamate degrading enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) decreases on moving from the latent phase (prior to seizures) to the chronic phase (after onset of seizures) in the kainate (KA) model of temporal lobe epilepsy. Western blotting and immunogold analysis of hippocampal formation indicated similar levels of GDH in the latent and chronic phases of KA injected rats and in corresponding controls. In contrast, the level of GS was increased in the latent phase compared with controls, as assessed by Western blots of whole hippocampal formation and subregions. The increase in GS paralleled that of glial fibrillary acidic protein (GFAP). Compared with the latent phase, the chronic phase revealed a lower level of GS (approaching control levels) but an unchanged GFAP content. The decrease in GS from latent to chronic phase was significant in whole hippocampal formation, dentate gyrus and CA3. It is concluded that kainate treated rats show an initial increase in GS, pari passu with the increase in GFAP, and a secondary decrease in GS that is not accompanied by a similar loss of GFAP. In a situation where glutamate catabolism is in high demand the secondary reduction in GS level may be sufficient to contribute to the seizure proneness that develops between the latent and chronic phases.

Immunocytochemical Evidence that Glutamate is a Neurotransmitter in the Cochlear Nerve: A Quantitative Study in the Guinea‐pig Anteroventral Cochlear Nucleus

The large so‐called type I afferents of the cochlear nerve carry the majority of the auditory input from the cochlea to the cochlear nuclei in the brainstem. These fibres are excitatory and previous studies have suggested they may use glutamate as their neurotransmitter. In the present investigation therefore, antibodies to glutamate and to the glutamate precursor, glutamine, were applied to resin sections of perfusion‐fixed brains and of in vitro brain slices subjected to depolarizing levels of potassium before fixation to study glutamate handling and synaptic release. Ultrathin sections were labelled by the immunogold technique, and the immunoreactivity was quantified by recording the density of gold particles over the various tissue profiles. Non‐primary, presumably inhibitory, terminals and glial processes were used as reference structures. The cochlear primary terminals proved to be strongly immunoreactive for glutamate. The density of glutamate labelling was higher in primary terminals than in non‐primary ones, and lowest in glial processes. The ratio between the mean glutamate and glutamine labelling densities was also higher in primary terminals than in non‐primary ones, and lowest in glial processes in each case. In the primary terminals, the glutamate immunoreactivity was higher over vesicle‐containing regions than over vesicle‐free regions, whilst glutamine was evenly distributed throughout. The in vitro brain slices showed a potassium‐induced, partly calcium‐dependent depletion of glutamate from the primary terminals but not from the non‐primary ones. These observations strongly support the conclusion that glutamate is a neurotransmitter of type I cochlear afferents.