Faculty Bibliography

The central nervous system of all vertebrate embryos is derived from a series of conspicuous segments, called neuromeres, that are particularly visible in the midbrain and hindbrain areas, giving rise to the brain stem sensory and motor nuclei. This article focuses on a series of eight embryonic rhombomeric segments whose progeny can be identified in adults by the locations of iteratively homologous reticulospinal neurons and cranial motor nuclei IV through XII. Evidence shows that these rhombomeric units represent domains of gene expression, lineage restriction, and accordingly, individual vestibular neuronal phenotypes with unique oculomotor and spinal projections. Preliminary electrophysiologic and behavioral correlates of a few vestibulo-oculomotor subgroups are used as examples to illustrate the hypothesis that homologous vestibular phenotypes likely exist in all taxa because the genetic prepattern is already well established in primitive vertebrates. Finally, the segmented hindbrain arrangement responsible for the longitudinally arranged column of vestibular subnuclei is placed in perspective with genetic and molecular approaches that will eventually permit a causal reconstruction of the signaling mechanisms responsible for the development of unique vestibular subgroups

Electrophysiological, ultrastructural, and immunohistochemical techniques were utilized to describe the excitatory and inhibitory vestibular innervation of extraocular motor nuclei in the goldfish. In antidromically activated oculomotor motoneurons, electrical stimulation of the intact contralateral vestibular nerve produced short-latency, variable amplitude electrotonic excitatory postsynaptic potentials (EPSPs) at 0.5-0.7 ms followed by chemical EPSPs at 1.0-1.3 ms. Stimulation of the ipsilateral vestibular nerve produced small amplitude membrane hyperpolarizations at a latency of 1.3-1.7 ms in which equilibrium potentials were slightly more negative than resting potentials. The inhibitory postsynaptic potentials (IPSPs) reversed with large amplitudes after the injection of chloride ions suggesting a proximal soma-dendritic location of terminals exhibiting high efficacy inhibitory synaptic conductances. In antidromically identified abducens motoneurons and putative internuclear neurons, electrical stimulation of the contralateral vestibular nerve produced large-amplitude, short-latency electrotonic EPSPs at 0.5 ms followed by chemical depolarizations at 1.2-1.3 ms. Stimulation of the ipsilateral vestibular nerve evoked IPSPs at 1.4 ms that were reversed after injection of current and/or chloride ions. gamma-Aminobutyric acid (GABA) antibodies labeled inhibitory neurons in vestibular subdivisions with axons projecting into the ipsilateral medial longitudinal fasciculus (MLF). Putative GABAergic terminals surrounded oculomotor, but not abducens, motoneurons retrogradely labeled with horseradish peroxidase. Hence the spatial distribution of GABAergic neurons and terminals appears highly similar in the vestibuloocular system of goldfish and mammals. Electron microscopy of motoneurons in the oculomotor and abducens nucleus showed axosomatic and axodendritic synaptic endings containing spheroidal synaptic vesicles establishing chemical, presumed excitatory, synaptic contacts with asymmetric pre- and/or postsynaptic membrane specializations. The majority of contacts with spheroidal vesicles displayed gap junctions in which the chemical and electrotonic synapses were either en face to dissimilar or adjacent to one another on the same soma/dendritic profiles. Another separate set of axosomatic synaptic endings, presumed to be inhibitory, contained pleiomorphic synaptic vesicles with symmetric pre- and/or postsynaptic membrane specializations that never included gap junctions. Excitatory and inhibitory synaptic contacts appeared equal in number but were more sparsely distributed along the soma-dendritic profiles of oculomotor as compared with abducens motoneurons. Collectively these data provide evidence for both disynaptic vestibular inhibition and excitation in all subdivisions of the extraocular motor nuclei suggesting the basic vestibulooculomotor blueprint to be conserved among vertebrates. We propose that unique vestibular neurons, transmitters, pathways, and synaptic arborizations are homologous structural traits that have been essentially preserved throughout vertebrate phylogeny by a shared developmental plan

Under normal physiological conditions, whole field visual motion generally occurs in response to either active or passive self-motion. In the laboratory, selective movement of the visual surround produces an optokinetic response (OKR) that acts primarily to support the vestibuloocular reflex (VOR). During visual world motion, however, the OKR can be viewed as operating independently over frequency and amplitude ranges insufficient for vestibular activation. The goal of the present study was to characterize this isolated behavior of the OKR in goldfish as an essential step for studying central neuronal correlates of visual-vestibular interactions and the mechanisms underlying oculomotor adaptation. After presentation of either binocular sinusoidal or step visual stimuli, conjugate eye movements were elicited with an amplitude and phase profile similar to that of other vertebrates. An early and a delayed component were measured with different dynamics that could be altered independently by visual training. The ensuing visuomotor plasticity was robust and exhibited five major characteristics. First, the gain of both early and delayed components of the OKR increased > 100%. Second, eye velocity decreased 0.5-2.0 s before the change in direction of stimulus velocity. Third, on lengthening the duration of a constant velocity visual stimulus (e.g., from 8 to 16 s), eye velocity decreased toward 0 degrees/s. This behavior was correlated with the direction and period as opposed to the frequency of the visual stimulus ('period tuning'). Fourth, visual stimulus training increased VOR eye velocity with a ratio of 0.6 to 1 to that measured for the OKR. Fifth, the OKR adaptation, eye velocity consistently oscillated in a conjugate, symmetrical fashion at 2.4 Hz in the light, whereas in the dark, a rhythmical low-amplitude eye velocity occurred at the visual training frequency. We conclude that the frequency and amplitude of visual stimuli for eliciting the goldfish OKR are well suited for complementing the VOR. Unlike most mammals, OKR adaptive modifications significantly alter VOR gain, whereas the effects of VOR training are much less on OKR gain. These observations suggest that both distributed circuits and discrete neuronal populations control visuo- and vestibulomotor performance. Finally, the existence of a rhythmic, 'period tuned' visuomotor behavior provides a unique opportunity to examine the neuronal mechanisms of adaptive plasticity

The discharge characteristics of Purkinje cells were analyzed in the goldfish cerebellum during eye movement and adaptation of the vestibulo-ocular reflex (VOR). Purkinje cells, identified by the simultaneous recording of complex and simple spikes, were recorded in the cerebellar area where electrical microstimulation elicited ipsiversive horizontal eye movements. Simple spikes of Purkinje cells displayed signals related to head and/or eye velocity as determined independently during either VOR suppression or optokinetic stimulation, respectively. Head velocity-only Purkinje cells (12%) increased their firing rate in relationship to ipsilateral head movements. Two types of eye velocity-only Purkinje cells (28%) were found that responded either in phase with ipsi- (16%) or contralateral (12%) eye movement, respectively. Purkinje cells combining both eye and head velocity (60%) were classified into two groups according to their preferred direction for eye movement. Eye velocity signals either added to (18%) or subtracted from (42%) head velocity during all visuo-vestibular interactions. Short term adaptive changes of the VOR were induced by oscillating goldfish in a moving visual surround that modified the ratio of eye to head velocity (gain) from a level of 1.0 (16 degrees/s) towards gains ranging from 2.5 (40 degrees/s) to -1.0 (-16 degrees/s). Simple spike modulation of individual Purkinje cells was shown to correlate well with VOR performance throughout adaptation irrespective of training direction. Purkinje cell behavior always equaled the algebraic summation of eye and head velocity signal sensitivity. Causality of signal generation was addressed by measuring Purkinje cell responses to both eye and head velocity separately throughout the time course of VOR adaptation. The sensitivity of each type of Purkinje cell was found to be independent of the VOR gain state. We therefore conclude that the changes responsible for short term VOR plasticity do not occur in the cerebellum. These observations suggest that Purkinje cells integrate corollary head and eye velocity signals to continuously adjust the set point of brainstem VOR interneurons that embrace the substantive site for adaptive plasticity.

This essay considers the ontogeny and phylogeny of the cranial neural circuitry producing rhythmic behaviors in vertebrates. These behaviors are characterized by predictable temporal patterns established by a neuronal network variously referred to as either a pacemaker, neural oscillator or central pattern generator. Comparative vertebrate studies have demonstrated that the embryonic hindbrain is divided into segmented compartments called rhombomeres, each of which gives rise to a distinct complement of cranial motoneurons and, as yet, unidentified populations of interneurons. We now propose that novel rhythmic circuits were innovations associated with the adoption of cardiac and respiratory pumps during the protochordate-vertebrate transition. We further suggest that the pattern-generating circuits of more recent innovations, such as the vocal, electromotor and extraocular systems, have originated from the same Hox gene-specified compartments of the embryonic hindbrain (rhombomeres 7-8) that gave rise to rhythmically active cardiac and respiratory circuits. Lastly, we propose that the capability for pattern generation by neurons originating from rhombomeres 7 and 8 is due to their electroresponsive properties producing pacemaker oscillations, as best typified by the inferior olive which also has origins from these same hindbrain compartments and has been suggested to establish rhythmic oscillations coupled to sensorimotor function throughout the neuraxis of vertebrates.

The distribution and dendritic morphology of neurons in the cat pretectal nuclear complex were analyzed with respect to their projection to the ipsilateral dorsal lateral geniculate nucleus (LGNd) and the ipsilateral inferior olive (IO). Single and double retrograde tracing techniques were combined with intracellular injections of either horseradish peroxidase into electrophysiologically identified pretectal neurons or Lucifer Yellow into retrogradely labeled somata. Pretectal cells afferent to the LGNd were located in the nucleus of the optic tract (NOT), adjacent dorsal terminal nucleus of the accessory optic system (DTN), and posterior pretectal nucleus (NPP). Cells projecting to the IO were also distributed throughout the NOT-DTN and dorsal part of the NPP. Separate tracer injections (fluorogold and horseradish peroxidase [HRP] or granular blue) into the LGNd and the IO showed considerable overlap of labeled neurons in the NOT and dorsal NPP. Double-labeled neurons, however, were not observed after double tracer injections into LGNd and IO. Partial topographical segregation of the two populations was observed along the dorsoventral axis because LGNd-projecting neurons exhibited maximum density ventral to that of IO neurons. Pretectal cells to the LGNd had cell body diameters between 16 and 48 microns. Somatic shapes varied between fusiform and multipolar with considerable overlap between these two morphological appearances. Neurons projecting to the IO exhibited similar cell body sizes and their morphology also varied from fusiform to multipolar. Quantitative analysis of dendritic field size and orientation, number and order of dendritic arborizations, and symmetry of the dendritic tree revealed no statistically significant difference between the two neuronal populations. Hence, neurons of the two populations cannot be unequivocally identified just from the dendritic morphology. By contrast, dendritic morphology was correlated with the topographical location of either cell type within the pretectal nuclei rather than projection. Thus, the morphological appearance of neurons located dorsally predominantly was fusiform while neurons located ventrally mostly were multipolar.