Zonal organization of flocculo-vestibular connections in rats.

Anatomical and electrophysiological evidence has contributed to the hypothesis that microzones in the mammalian flocculus are organized to reflect control of eye movements in the planes of semicircular canals. Adult male Long-Evans rats received iontophoretic injections of FluoroGold and/or tetramethylrhodamine dextran amine (10,000 molecular weight, "FluoroRuby") into the vestibular nuclei. The distribution of retrogradely labeled Purkinje cells revealed that efferent projections from the dorsal surface of the flocculus and the ventral paraflocculus to the superior vestibular nucleus, rostral medial vestibular nucleus, ventral lateral vestibular nucleus, and caudal aspect of the vestibular nuclear complex (caudal medial vestibular nucleus, inferior vestibular nucleus and nucleus prepositus hypoglossi) tended to correspond to previously identified climbing fiber zones [Ruigrok et al. (1992) J. comp. Neurol. 316, 129-150] in a manner consistent with other mammals. However, vestibular nucleus projections from the ventral surface of the flocculus did not appear to respect climbing fiber zonal boundaries. Rather, climbing fiber zones each contained interdigitated groups of Purkinje cells that project to different vestibular nuclear regions. It is suggested that this pattern of flocculus efferent organization is a specialization for controlling the activity of primary and accessory extraocular muscle pairs to confine vestibulo-ocular reflexes within semicircular canal planes when the "center of regard" is located at different eccentricities.

Unipolar brush cell: a potential feedforward excitatory interneuron of the cerebellum.

Unipolar brush cells are a class of interneurons in the granular layer of the mammalian cerebellum that receives excitatory mossy fiber synaptic input in the form of a giant glutamatergic synapse. Previously, it was shown that the unipolar brush cell axon branches within the granular layer, giving rise to large terminals. Single mossy fiber stimuli evoke a prolonged burst of firing in unipolar brush cells, which would be distributed to postsynaptic targets within the granular layer. Knowledge of the ultrastructure of the unipolar brush cell terminals and of the cellular identity of its postsynaptic targets is required to understand how unipolar brush cells contribute to information processing in the cerebellar circuit. To investigate the unipolar brush cell axon and its targets, unipolar brush cells were patch-clamped in fresh parasagittal slices from rat cerebellar vermis with electrodes filled with Lucifer Yellow and Biocytin, and examined by confocal fluorescence and electron microscopy. Biocytin was localized with diaminobenzidine chromogen or gold-conjugated, silver-intensified avidin. Light microscopic examination revealed a single thin axon emanating from the unipolar brush cell soma that gave rise to 2-3 axon collaterals terminating in mossy fiber-like rosettes in the granular layer, typically within a few hundred microm of the soma. In some cases, axon collaterals crossed the white matter within the same folium before terminating in the adjacent granular layer. Electron microscopic examination of serial ultrathin sections revealed that proximal unipolar brush cell axons and axon collaterals were unmyelinated and devoid of synaptic contacts. However, the rosette-shaped enlargements of each collateral formed the central component of glomeruli where they were surrounded by dendrites of granule cells and/or other unipolar brush cells, with which they formed asymmetric synaptic contacts. A long-latency repetitive burst of polysynaptic activity was observed in granule cells in this cerebellar region following white matter stimulation. The unipolar brush cell axons, therefore, form a system of cortex-intrinsic mossy fibers. The results indicate that synaptic excitation of unipolar brush cells by mossy fibers will drive a large population of granule cells, and thus will contribute a powerful form of distributed excitation within the basic circuit of the cerebellar cortex.

Organization of the coeruleo-vestibular pathway in rats, rabbits, and monkeys.

Inputs from locus coeruleus (LC) appear to be important for altering sensorimotor responses in situations requiring increase vigilance or alertness. This study documents the organization of coeruleo-vestibular pathways in rats, rabbits and monkeys. A lateral descending noradrenergic bundle (LDB) projects from LC to the superior vestibular nucleus (SVN) and rostral lateral vestibular nucleus (LVN). A medial descending noradrenergic bundle (MDB) projects from LC to LVN, the medial vestibular nucleus (MVN), group y and rostral nucleus prepositus hypoglossi (rNPH). There is a characteristic, specific pattern of innervation of vestibular nuclear regions across the three species. A quantitative analysis revealed four distinct innervation density levels (minimal, low, intermediate and high) across the vestibular nuclei. The densest plexuses of noradrenergic fibers were observed in the SVN and LVN. Less dense innervation was observed in the MVN, and minimal innervation was observed in the inferior vestibular nucleus (IVN). In monkeys and rabbits, rostral MVN contained a higher innervation density than the rat MVN. In monkeys, the rNPH also contained a dense plexus of fibers. Selective destruction of terminal LC projections (distal axons and terminals) by the neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) resulted in a dramatic reduction of immunoreactive fibers within the vestibular nuclear complex of rats, suggesting that the source of these immunoreactive fibers is LC. Retrograde tracer injections into the vestibular nuclei resulted in labeled cells in the ipsilateral, caudal LC and adjacent nucleus subcoeruleus. It is hypothesized that the regional differences in noradrenergic innervation are a substrate for differentially altering vestibulo-ocular and vestibulo-spinal responses during changes in alertness or vigilance.

N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) has differential efficacy for causing central noradrenergic lesions in two different rat strains: comparison between Long-Evans and Sprague-Dawley rats.

We tested the hypothesis that Long-Evans (LE) and Sprague-Dawley (SD) rat strains were equally sensitive to the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) with respect to central lesions of locus coeruleus (LC) terminals as measured by immunohistochemical localization of dopamine-beta-hydroxylase (D beta H). Analysis of D beta H immunoreactivity was made by both qualitative and quantitative methods. Intraperitoneal injections of 50 mg/kg DSP-4 caused a dramatic reduction of noradrenergic terminals in the neocortex, hippocampus and cerebellum of SD, but not LE rats as compared to saline-injected controls. This finding indicates that LE rats are less sensitive than SD rats to the neurotoxic effects of DSP-4 in the central nervous system.

Evidence for a noradrenergic projection to the subcommissural organ.

Several physiological studies have shown that the subcommissural organ (SCO) is influenced by catecholamines. This study provides immunohistochemical evidence for a noradrenergic input to the SCO of rats. A light plexus of tyrosine hydroxylase (TH)-and dopamine-beta-hydroxylase (D beta H)-positive axons present in the SCO of both Long-Evans and Sprague-Dawley rats. The innervation density was greatest in the hypendymal wing of the rostral aspect of the SCO and it declined both caudally in the hypendymal wing and medially in the hypendymal layer. Some TH- and D beta D beta H-immunoreactive fibers entered the lateral margin of the ependymal layer along the basal surface of ependymal cells; others coursed medially in the transverse plane to ramify along the base of the ependymal cells. These fibers are presumed to be noradrenergic because phenylethanolamine N-methyltransferase immunoreactivity was absent in adjacent sections through the SCO. Considering the potential role of the SCO region in sodium homeostasis, these data suggest that central noradrenergic input to the SCO may parallel peripheral catecholaminergic mechanisms that regulate sodium balance.

Immunohistochemical demonstration of regionally selective projections from locus coeruleus to the vestibular nuclei in rats.

This study describes a regionally selective projection of tyrosine hydroxylase and dopamine beta-hydroxylase-immunoreactive fibers from locus coeruleus (LC) and the A4 region of nucleus subcoeruleus to the vestibular nuclear complex in Long-Evans and Sprague-Dawley rats. These fibers travel in two distinct pathways. A lateral descending noradrenergic bundle provides input from LC to the superior vestibular nucleus (SVN), the cochlear nuclei, and the cerebellar cortex. A medial descending noradrenergic bundle provides input to the lateral vestibular nucleus (LVN), medial vestibular nucleus (MVN), and the inferior vestibular nucleus (IVN) before continuing on to the cochlear and cerebellar nuclei. The terminal plexus of these fibers varies markedly across these vestibular nuclear regions. Immunoreactive axons form a dense plexus around somata and proximal dendrites of Deiters' neurons in dorsal LVN. The axon plexus is less dense in SVN and ventral LVN, and relatively sparse in MVN and IVN. This regional selectivity of noradrenergic innervation suggests that central adrenergic systems may selectively modulate vestibulospinal reflexes at the level of the vestibular nuclear complex.

Retinal direction-sensitive input to the accessory optic system: an in vitro approach with behavioral relevance.

Retinal application of gamma-aminobutyric acid (GABA) antagonists block direction-sensitive (DS) responses in turtle in two ways: (1) the selectivity of DS retinal ganglion cells in vitro, and (2) the eye's ability to track the direction of full field image motion. The experiments described below demonstrate that an important locus for retinal slip computation by the accessory optic system (AOS) occurs in the retina. Visual responses were measured physiologically and behaviorally from turtles which had their telencephalon removed. Physiological responses to visual field movement were recorded in the AOS using an in vitro brain preparation. DS responses of single cells were blocked by intravitreal application of bicuculline. The behavioral approach was to measure optokinetic nystagmus (OKN) in lesioned animals. OKN occurred in the absence of the telencephalon, yet was disrupted following an intravitreal injection of bicuculline. Thus, both experimental approaches showed that DS processing exists without the telencephalon, yet is disrupted by GABA antagonists applied to the retina.