Connections of Purkinje cell axons of lobule X with vestibulocerebellar neurons projecting to lobule X or IX in the rat

Connections of Purkinje cell axons of lobule X (nodulus) with vestibulocerebellar neurons projecting to lobule X or IX (uvula) were revealed in the rat. Purkinje cell axons were anterogradely labeled with biotinylated dextran (BD) injected into sublobule Xa while vestibular neurons were retrogradely labeled with cholera toxin subunit B (CTB) injected into sublobule Xa or IXc. Labeled terminals of Purkinje cell axons of lobule X were numerous in the superior vestibular nucleus (SV), medial parts of the parvocellular (MVpc) and the caudal part (MVc) of the medial vestibular nucleus (MV), and group y. These subdivisions of the vestibular nuclei contained many neurons projecting to lobule X or IX. Lobule-X-projecting and lobule-IX-projecting neurons were in contact with terminals of Purkinje cell axons of lobule X in the MVpc and MVc. They were distributed dorsally to medially in medial parts of the MVpc and MVc. The present study suggests that Purkinje cells in lobule X regulate the output of a population of lobule-X-projecting or lobule-IX-projecting neurons of the MVpc and MVc.     

Receptive field organization of climbing fiber afferents responding to optokinetic stimulation in the cerebellar nodulus and flocculus of the pigmented rabbit

Summary and conclusions Under anesthesia with N₂O (70%) and halothane (2–4%), complex spikes of Purkinje cells were extracellularly recorded in the nodulus and flocculus of immobilized pigmented rabbits. Optokinetic stimulation (OKS) was delivered to each eye as repetitive movements of a random dot pattern. The visual field of each eye was divided into anterior, central and posterior fields at axes 45° and 135° along the horizon. With OKS of the ipsilateral eye, the preferred direction of complex spike responses was: (1) forward (F) in all visual fields (F response), (2) upward (U) in both the anterior and central visual fields but downward (D) in the posterior visual field (U response), or (3) no response (N) in any of the visual fields (N response). With OKS of the contralateral eye, the preferred direction was: (1) backward (B) in both the anterior and central visual fields but N in the posterior visual field (B response), (2) U in the anterior but D in both the central and posterior visual fields (D response), or (3) N in all visual fields. Purkinje cells were classified into five categories in terms of the complex spike responses to OKS to the central visual fields of the ipsi-/contralateral eyes: F/B, F/N, U/D, U/N and N/D types. In cells with ipsi-F and/or contra-B responses, OKS delivered above the horizon induced F and/or B responses, but OKS below the horizon induced no response. In cells with contra-D response, OKS both above and below the horizon induced D responses. Cells with ipsi-U responses showed unusually complex direction selectivity: in the anterior, central and posterior fields, the preferred direction was U, U and D, respectively, above the horizon, as opposed to N, D and D below the horizon. In the nodulus, F/B and F/N type cells were localized in the ventral lamella within two distinct longitudinal zones about 0.5–1.5 and 2.5–3.5 mm from the midline, while U/D, U/N and N/D type cells were found in both the ventral and dorsal lamellae within a longitudinal zone about 1.5–2.5 mm from the midline. In the flocculus, the locations of F/N (or F/B), U/N and N/D type cells roughly corresponded to H, anterior V and R zones, respectively, as previously reported on the basis of the direction of eye movements induced by microstimulation. The receptive field of visual climbing fiber inputs to the nodulus and flocculus is organized such that complex spike activity is best modulated with retinal image slips caused by head rotation around the axis of either the horizontal (F/B, F/N types) or the anterior canal (U/D, U/N, N/D types) on the ipsilateral side.     

Optokinetic response of simple spikes of Purkinje cells in the cerebellar flocculus and nodulus of the pigmented rabbit

Summary Under anesthesia with N₂O (70%) and halothane (2–4%), Purkinje cell activities were extracellularly recorded in the flocculus and nodulus of immobilized pigmented rabbits. Large field (60° × 60°) optokinetic stimulation (OKS) was delivered to the central visual field of one eye with a constant velocity (0.1–4.0 °/S) at 0°, 45°, 90° or 135° to the horizontal plane of the eye. Most of the Purkinje cells in the flocculus and the nodulus showed significant simple spike modulations to OKS delivered to either eye. As a whole, the preferred directions of simple spike responses in the flocculus had the same orientation as those of complex spike responses. However, the preferred directions and amplitudes of modulation of simple spike responses did not necessarily correlate with those of complex spike responses in individual flocculus Purkinje cells. On the other hand, the preferred directions of simple and complex spike responses were not necessarily in the same orientation in the nodulus. The optimum velocity for simple spike responses was in the range 0.1–2.0°/s for Purkinje cells in both the flocculus and the nodulus. The amplitude and time to peak of the simple spike responses of nodulus Purkinje cells were significantly smaller and longer, respectively, than those of flocculus Purkinje cells. In both the flocculus and the nodulus, Purkinje cells whose simple spikes preferred the horizontal orientation (H cells) and the vertical orientation (V cells) showed clustering. In particular, zonal organization was noted in the flocculus. H cells were localized in a dorso-ventral zone in the rostral one third of the flocculus, and V cells were in two distinct zones rostral and caudal to the H cell zone. The locations of H and V cells in the flocculus correspond to the H zone and V zones, respectively, determined on the basis of the preferred directions of complex spike responses to OKS. This indicates that the same subdivisions of the flocculus are supplied with optokinetic signals with the same orientation selectivity through both mossy and climbing fibers, and suggest that such subdivisions of the flocculus are functional units which control horizontal and vertical components of optokinetic eye movements. The present results indicate that the flocculus and the nodulus are supplied with distinct optokinetic signals through mossy fibers and play different roles in controlling optokinetic eye movements.     

The pontine projection to the flocculonodular lobe and the paraflocculus studied by means of retrograde axonal transport of horseradish peroxidase in the rabbit

Summary The occurrence and distribution of labeled cells in the pontine nuclei were mapped following injections of small amounts of horseradish peroxidase (0.05–0.5 l, 50% suspension) in the flocculus, nodulus and the dorsal and ventral paraflocculus in adult albino rabbits. While no labeled cells were found in the pontine nuclei following injections in the nodulus, some were present following injections in the flocculus and a great number following injections in the paraflocculus. The projections onto the flocculus and paraflocculus are precisely organized. Following injections in the paraflocculus labeled neurons are arranged in four columns (E and G in the paramedian pontine nucleus, F in the peduncular and H in the dorsolateral nucleus). Following injections in the ventral paraflocculus labeled cells are present only in parts of column E and F, while columns G and H and parts of E and F project onto the dorsal paraflocculus. Following injections in the flocculus labeled cells occur in the rostral part of column E only.A comparison between the sites of termination of pontine afferents and the areas giving origin to floccular and parafloccular fibers shows that only few fibers mediating visual impulses end in these pontine areas, while they receive numerous fibers from gyrus cinguli and areas 18 and 19 of the cerebral cortex.     

Diminution and reversal of eye movements induced by local stimulation of rabbit cerebellar flocculus after partial destruction of the inferior olive

Summary After the dorsal cap and adjacent ventrolateral outgrowth regions of the inferior olive had been chronically destroyed in the rabbits, the eye movements evoked by local stimulation of the flocculus were reduced in amplitude and reversed in direction, indicating that the inhibition by flocculus Purkinje cells of vestibulo-ocular relay neurons could no longer be actuated by the stimulation.M. Dufossé was supported by French-Japanese Cultural Exchange Programs.This work was supported by a grant from the Educational Ministry of Japan (212103)     

Cholinergic modulation of optokinetic and vestibulo-ocular responses: a study with microinjections in the flocculus of the rabbit

Summary In spite of a large body of histochemical evidence for a cholinergic system in the cerebellum, particularly in lobules IX and X, the physiological role of such a system has remained obscure. In view of the important role of these same lobules in the control of the vestibulo-ocular (VOR) and optokinetic (OKR) responses, we tested the effect of microinjections of cholinergic (ant)agonists in the flocculus of the rabbit on these reflexes. Very marked effects were found. Bilateral floccular injection of the aspecific cholinergic agonist carbachol raised the gain of the OKR by about 0.46 above the baseline values, while the gain of the VOR in darkness was raised by about 0.14. These effects were statistically significant and persisted for several hours. Similar, but smaller effects were obtained after injection of eserine, an inhibitor of acetylcholinesterase. Thus, the effects could be produced by increasing the naturally present amount of acetylcholine. Microinjections of the nicotinic blocker mecamylamine reduced the gain of the VOR and OKR, although these effects did not reach statistical significance. The muscarinic blocker atropine significantly reduced the gain of the OKR, but not of the VOR. The present results argue strongly for an important physiological role of the cholinergic system in the cerebellum. Specifically, acetylcholine appears to be involved in the modulation of oculomotor reflexes through the flocculus.     

Branching projection of the nucleus “k” neurons to the rabbit cerebellar paramedian lobule: a retrograde fluorescent tracing study

The nucleus "k", in the reticular core of the rabbit caudal pons, is divided into a large medial (composed of dorsal k1 and ventral k2) and a small lateral (k3) subdivision. In this study, the nucleus "k" subdivisions were examined in the rabbit with respect to projections to the cortex of rostral (rPML; face-forelimb region) and caudal (cPML; hindlimb region) paramedian lobule of the cerebellum. The retrograde fluorescent labeling method with Fast Blue (FB) and Diamidino Yellow (DY) was used. Numerous single FB or DY labeled neurons were found in defined regions of all nucleus "k" subdivisions bilaterally, with an ipsilateral preponderance. The distribution of these neurons indicated that afferents originating from different nucleus "k" subdivisions terminated in overlapping regions within the rPML and the cPML rather than in separate domains. Apart from this, double FB + DY labeled neurons (n = 104) were intermingled within a common region of single labeling, but exclusively on the ipsilateral side. Such neurons occupied predominantly the central and lateral regions of the caudal two thirds of the k1 subdivision, and were scattered in the caudal half of k2 as well as throughout the entire rostrocaudal extent of the k3 subdivision. The size of labeled perikarya varied from 20 to 40 microm in diameter. The number of neurons with branching axons was considerably lower than those with single projections to the rPML and the cPML. It amounted to about 3% in k1 and k3, and 2% in the k2 subdivision. However, this population may form an intralobular link between two somatotopically non-corresponding PML regions. The present study provides a morphological basis for further investigations for comparison with other species using both anatomical and electrophysiological methods, also with respect to other connections of the nucleus "k".     

Commissural and intrinsic connections of the vestibular nuclei in the rabbit: a retrograde labeling study

Summary The intrinsic and commissural projection of the vestibular nuclei were investigated by means of retrograde transport of normal (HRP) and wheatgerm-agglutinated horseradish peroxidase (WGA-HRP). It was found that within each vestibular complex, the superior (SV), medial (MV) and descending (DV) vestibular nuclei are reciprocally connected. A rostrocaudally oriented column of medium-sized and large neurons, comprising the central SV and the magnocellular MV (MVmc) receives input from the surrounding neurons and does not reciprocate this projection. Efferents from group y terminate in the SV, MV and DV. The infracerebellar nucleus (INF) as well as the interstitial nucleus of the VIII the nerve (IN) supply fibers to the MV and DV. The neurons that participate in the commissural projection are distributed throughout the vestibular complex with the exception of the lateral vestibular nucleus (LV) and group x. The largest number of cells was found in the MV. The HRP labeled cells show a tendency to cluster into rostrocaudally oriented groups. Each nucleus projects to more than one contralateral nucleus. Group y shows a more extensive contralateral projection than the bordering INF. It was concluded that quantitative differences in connectivity were present between a core region in the vestibular complex and peripheral parts. This core region comprises the central SV, the LV, the MVmc and extends into the rostral DV. It receives predominantly intrinsic input from the surrounding vestibular neurons and is in contrast to these latter neurons only minimally involved in the commissural projection.Abbreviations AChE acetylcholinesterase - bc brachium conjunctivum - bp brachium pontis - CE nucleus cuneatus externus - CO nuclei cochlearis - cr corpus restiforme - DV nucleus vestibularis descendens - DX nucleus dorsalis vagi - F nucleus fastigii - flm fasciculus longitudinalis medialis - gVII genu of the nervus facialis - group x, y, f groups x, y and f of Brodal - HRP horseradish peroxidase - IA nucleus interpositus anterior - IN nucleus interstitialis of nVIII - INF nucleus infracerebellaris - L nucleus lateralis - LV nucleus vestibularis lateralis - flm fasciculus longitudinalis medialis - MV nucleus vestibularis medialis - MVc caudal MV - MVmc magnocellular MV - MVpc parvocellular MV - nV nervus trigeminus - nVI nervus abducens - nVII nervus facialis - NV par nucleus vestibularis parabrachialis - PH nucleus prepositus hypoglossi - rV ramus descendens of nV - S nucleus and tractus solitarius - sad stria acustica dorsalis - SV nucleus vestibularis superior - tu tractus uncinatus - VI nucleus abducens - VM nucleus masticatorius - VOR vestibulo-ocular reflex - VP nucleus princeps trigemini - WGA-HRP wheatgerm-agglutinated HRP - XII nucleus hypoglossus     

Vestibular nucleus projections to nucleus tractus solitarius and the dorsal motor nucleus of the vagus nerve: potential substrates for vestibulo-autonomic interactions

Autonomic effects of vestibular stimulation are important components of phenomena as diverse as acute vestibular dysfunction and motion sickness. How ever, the organization of neural circuits mediating these responses is poorly understood. This study presents evidence for direct vestibular nucleus projections to brain stem regions that mediate autonomic function. One group of albino rabbits received injections of Phaseolus vulgaris leucoagglutinin into the vestibular nuclei. The tracer was visualized immunocytochemically with standard techniques. Anterogradely labeled axons from the caudal medial vestibular nucleus (cMVN) and inferior vestibular nucleus (IVN) could be traced bilaterally to nucleus tractus solitarius (NTS). Fewer axons ended near the somata of neurons in the dorsal motor nucleus of the vagus nerve (DMX). A second group of rabbits received pressure or iontophoretic injections of cholera toxin B-HRP or Fluoro-Gold into a region including NTS and DMX. Retrogradely labeled neurons were observed bilaterally in the caudal half of cMVN and ipsilaterally in IVN. The labeled somata were small and they tended to occupy the center of cMVN in transverse sections. These previously unreported vestibular nucleus projections to NTS and DMX are a potential substrate for vestibular influences on autonomic function. In particular, they may contribute to both cardiovascular control during head movements (e.g., orthostatic reflexes) and autonomic manifestions of vestibular dysfunction, motion sickness and exposure to altered gravitational environments.     

Convergence of cortico- and cuneopontine projections onto components of the pontocerebellar system in the rat: an anatomical and electrophysiological study

Summary Previous studies in the rat have demonstrated that corresponding peripheral tactile and somatosensory cortical inputs converge within the granule cell layer of various cerebellar lobules and further that descending corticopontine projections from the forelimb sensory cortex (FLSCx) partially overlap with the projection zones of ascending basilar pontine afferents from nucleus cuneatus (NC). The present study employed anatomical and electrophysiological procedures to determine whether cortical and dorsal column nuclear afferent projections converge on pontine neurons that, in turn, provide mossy fiber input to the granule cell layer of the paramedian lobule (PML), i. e., that portion of the rodent cerebellum shown to receive forelimb peripheral inputs. The combination of the orthograde and retrograde axonal transport of horseradish peroxidase (HRP) conjugated to wheat germ agglutinin (WGA) was used light microscopically to demonstrate that orthogradely labeled projections from injections of the FLSCx and NC converged with ponto-paramedian projection neurons that were retrogradely labeled from injections of the PML. These studies were also repeated in conjunction with ablations of either the FLSCx or NC which resulted in the ultrastructural identification of degenerating, as well as WGA-HRP labeled axonal boutons of these pontine afferent projections thus confirming that such projections actually formed synaptic contacts with the retrogradely labeled pontoparamedian projection neurons. Single unit recording analyses of neurons in the ventromedial region of the basilar pons following combined electrical stimulation of various regions of the sensorimotor cortex and the contralateral body surface indicated that approximately 40% of all cells recorded responded to electrical stimulation of corresponding regions of the cortex and periphery, particularly the FLSCx and the forepaw. Natural cutaneous stimuli applied to the forepaw that also elicited responses in these same groups of basilar pontine neurons and were associated with relatively small receptive fields. Taken together, these observations indicate that the previously observed convergence of peripheral and somatosensory cortical inputs within the granule cell layer of the cerebellar cortex may be at least partially organized at the level of the basilar pons.     

A study of branching in the projection from the inferior olive to the x and lateral c1 zones of the cat cerebellum using a combined electrophysiological and retrograde fluorescent double-labelling technique

Summary The pattern of transverse branching in the olivocerebellar projection to the x zone in the vermis and the lateral c₁ zone in the paravermis of the cat anterior lobe was studied using a combined electrophysiological and retrograde double-labelling tracer technique. Fluorochrome-tagged latex microspheres were well suited for this purpose. The results show that the region of olive that supplies climbing fibres to the two zones forms a continuous, rostrocaudally directed column about 2.25 mm in length, in a caudo-lateral to rostromedial part of the medial accessory olive (MAO), on average between A-P levels 12.50-10.50. This column may be divided into caudal and rostral halves that project respectively to the x and lateral c₁ zones in the apical folia of lobules V/VIa. Partial overlap between these two territories occurs in an intermediate region (A-P levels 12.00-11.00) in middle MAO where olive cells that supply climbing fibres to either x or lateral c₁ are intermingled with a smaller population of cells whose axons branch to provide climbing fibres to both zones. Quantitative analysis showed that, when different tracers were injected into each zone in the same animal, double-labelled cells represented only 5–7% of either single-labelled cell population within this area of overlap. It is concluded that, although some transverse branching is present within the olivocerebellar projection to the x and lateral c₁ zones in the apical folia of lobule V, such branching is not extensive.     

Network organization of the connectivity between parietal area 7, posterior cingulate cortex and medial pulvinar nucleus: a double fluorescent tracer study in monkey

Summary The topographic distribution of medial pulvinar neuronal populations projecting to area 7a and to posterior cingulate gyrus (area 23) was investigated with retrograde axonal transport of fluorescent dyes. In an initial stage, separate injections of fast blue and diamidino-yellow were placed in area 7a. Two segregated backfilled cell populations were observed in the cingulate gyrus revealing a topographic distribution of cortico-cortical connections. In a second stage, the two dyes were injected in area 7a and in the posterior cingulate gyrus. After injections in two cortical sub-areas previously shown to be in topographic correspondence, the two projecting populations observed in the medial pulvinar over-lapped extensively. In the case of injections in two regions not topographically correspondent, the two medial pulvinar populations were found to be segregated. These findings reveal that within both posterior parietal cortex and posterior cingulate gyrus, sub-areas which are in topographic correspondence receive projections from a common region of the medial pulvinar.     

Collateralization of cerebellar efferent projections to the paraoculomotor region,superior colliculus,and medial pontine reticular formation in the rat: a fluorescent double-labeling study

Summary Collateralization of cerebellar efferent projections to the oculomotor region, superior colliculus (SC), and medial pontine reticular formation (mPRF) was studied in rats using fluorescent tracer substances. In one group, True Blue (TB) was injected into the oculomotor complex (OMC), including certain paraoculomotor nuclei and supraoculomotor ventral periaqueductal gray (PAG), and Diamidino Yellow (DY) was injected into the medial pontine reticular formation (mPRF) or pontine raphe. The largest number of single-TB-labeled (paraoculomotor-projecting) cells was observed in the medial cerebellar nucleus (MCN) and posterior interposed nucleus (PIN), whereas the largest number of single-DY-labeled (mPRF-projecting) cells was in the MCN. Double-TB/DY-labeled cells were present in the caudal two-thirds of the MCN, suggesting that some MCN neurons send divergent axon collaterals to the paraoculomotor region and mPRF. In another group, TB was injected into the SC and DY into the mPRF. The largest number of single-TB-labeled (SC-projecting) cells was in the PIN, although a considerable number of cells was observed in the caudal MCN, and ventral lateral cerebellar nucleus (LCN). Single-DY-labeled (mPRF-projecting) neurons were primarily located in the central and ventral MCN, but were also present in the lateral anterior interposed (AIN) and in the LCN. Double-TB/DY-labeled neurons were observed in the caudal two-thirds of the MCN and in the central portion of the LCN. The most significant new findings of the study concerned the MCN, which not only contained neurons that projected independently to the paraoculomotor region, SC, and mPRF, but also contained a considerable number of cells which collateralized to project to more than one of these nuclei. The possibility that the MCN projects to the supraoculomotor ventral PAG (containing an oculomotor interneuron system) and to the mPRF, which in the cat and monkey contain neural elements essential to the production of saccadic eye movements, is discussed. The anatomical findings suggest that the MCN in the rat plays an important role in eye movement.Abbreviations AI anterior interposed nucleus - AIN anterior interposed nucleus - BC brachium conjunctivum (sup. cerebellar peduncle) - dlh dorsolateral hump of the AI - dmc dorsomedial crest of the AI - IC inferior colliculus - ICP inferior cerebellar peduncle - Inf infracerebellar nucleus - L lateral cerebellar (dentate) nucleus - LCN lateral cerebellar (dentate) nucleus - M medial cerebellar (fastigial) nucleus - MCN medial cerebellar (fastigial) nucleus - MLF medial longitudinal fasciculus - mPRF medial pontine reticular formation (incl. nuc. reticularis pontis oralis and caudalis) - OMC oculomotor complex - OMN oculomotor nucleus - PI posterior interposed nucleus - PIN posterior interposed nucleus - RN red nucleus - SC superior colliculus - Vl lateral vestibular nucleus - Vs superior vestibular nucleus - Y cell group Y     

The accessory optic system of the rabbit, cat, dog and monkey: a whole-mount HRP study

Summary The three-dimensional fiber pathways of the accessory optic system in the rabbit, cat, dog and monkey were studied in whole-mounted preparations of the diencephalon and the midbrain, without sectioning, by anterograde labeling of retinal axons with horseradish peroxidase (HRP). HRP histochemical studies on alternate serial coronal sections were also performed. The rabbit accessory optic system exhibited two fasciculi (the inferior fasciculus, and the superior fasciculus consisting of the anterior fibers and the posterior fibers) and three terminal nuclei (the medial terminal nucleus, and the anterior and posterior porcions of the lateral terminal nucleus), but lacked the dorsal terminal nucleus. In the cat and dog, only the posterior fibers of the superior fasciculus were detected. The inferior fasciculus and the anterior fibers of the superior fasciculus were absent. The medial terminal nucleus and the posterior portion of the lateral terminal nucleus were commonly observed in the cat and dog. The cat accessory optic system possessed the dorsal terminal nucleus, and the dog accessory optic system possessed the anterior portion of the lateral terminal nucleus. The monkey (Macaca fuscata) accessory optic system consisted of the posterior fibers of the superior fasciculus, but blacked the inferior fasciculus and the anterior fibers of the superior fasciculus. Most of the posterior fibers terminated in the well-developed posterior portion of the lateral terminal nucleus located on the upper surface of the cerebral peduncle. A small number of posterior fibers projected to the poorly-developed medial terminal nucleus. Based on these findings, species differences in the mammalian accessory optic system were discussed.     

Thalamic inputs to the rabbit visual cortex: Identification and organization using horseradish peroxidase (HRP)

Summary Horseradish peroxidase (HRP) has been injected in visual cortical area 1 (V1, striate cortex) of 33 rabbits (16 received a unilateral injection, 17 bilateral injections) in order to identify its thalamic inputs and to determine their retinotopic organization. This study has shown that when HRP is injected into different portions of V1: 1. Labeled cells are consistently seen in the dorsal lateral geniculate nucleus (LGNd) and are always organized into horizontal columns arranged perpendicularly to the long axis of the LGNd; although the columns are always present in the alpha sector of the LGNd, they extend into the beta sector of this nucleus only occasionally. 2. With a series of injection sites that lie on or near the medial edge of V1 and run rostrocaudally, cell columns shift from ventromedial to dorsolateral within the LGNd. With a series of injection sites that are located along the lateral border of V1 and run from rostral to caudal, columns of labeled cells move from ventral to dorsal along the medial edge of the LGNd. 3. In some of the experiments, with injections in either medial or lateral portions of V1, columns of HRP-labeled cells have also been found within the pulvinar.A preliminary report of this study (Karamanlidis and Giolli, 1976) has been presented at the sixth annual meeting of the Society for Neurosciences, Toronto, Canada, November 1976     

Axonal branching of medullary swallowing neurons projecting on the trigeminal and hypoglossal motor nuclei: demonstration by electrophysiological and fluorescent double labeling techniques

Summary The projections of ventral medullary reticular neurons on both trigeminal (Vth) and hypoglossal (XIIth) motor nucleus were studied in sheep anesthetized with halothane. In a first series of experiments, extracellular microelectrodes were used to record the activity of medullary swallowing interneurons (SINs) located in the ventral region (around the nucleus ambiguus) of the swallowing center. Antidromic activation after electrical stimulation of the Vth and XIIth nuclei was tested in 83 SINs. For 38 SINs a clear antidromic activation was observed and for 8 of them the response was triggered by stimulation of either nucleus. As confirmed by the reciprocal collision test, these 8 SINs had branched axons sending information to both nuclei tested. Average latencies for antidromic activation of branched SINs after stimulation of the XIIth and the Vth motor nucleus were 2.2±0.6 ms and 2.7±0.8 ms respectively. The axonal conduction velocity of these neurons was 4.4±1.3 m/s for the collateral to the Vth motor nucleus and 2.7±0.7 m/s for axons projecting to the XIIth motor nucleus. In a second series of experiments the double retrograde labeling technique was used to confirm the existence of neurons with branched axons in the medullary regions corresponding to the swallowing center. Small and well localized injections of Fast Blue (FB) and Diamidino Yellow (DY) fluorescent tracers were made in the Vth and in the XIIth motor nucleus respectively. A relatively large number of double-labeled cells was found in the ventral region of swallowing center (reticular formation around the nucleus ambiguus, 2–4 mm in front of obex). Such neurons (supplying both the XIIth and the Vth motor nucleus) appeared mixed with those innervating only either the XIIth or the Vth motor nucleus. Each type of neuron, i.e. single or double labeled, was shown to have bilateral distribution with an ipsilateral predominance.     

Glutamate-like immunoreactivity in synaptic terminals of the posterior cingulopontine pathway: a light and electron microscopic study in the rabbit

A postembedding immunoperoxidase method for light microscopy was used to localize glutamate-like immunoreactivity in the rabbit basilar pontine nuclei. Labelled fibre bundles, neuronal cell bodies and numerous puncta of diverse size were heavily glutamate immunoreactive throughout all subdivisions of the pontine nuclei. To determine whether some of the glutamate-immunoreactive puncta were synaptic terminals of posterior cingulate cortical neurons, a double-labelling technique involving an anterograde tract-tracing method and a postembedding immunogold procedure for electron microscopy was used. A quantitative evaluation of gold particle densities revealed that anterogradely labelled cingulopontine synaptic terminals were about twice as immunoreactive as their postsynaptic dendrites, perikaryal and glial profiles and about three times more than symmetric synaptic terminals. The present results indicate that the posterior cingulopontine projection contains high levels of glutamate at its synaptic terminals. This observation provides further support to the role for glutamate as a neurotransmitter in the corticopontine pathway.     

A comparison of the distribution of the cerebellar and cortical connections of the nucleus of Darkschewitsch (ND) in the cat: a study using anterograde and retrograde HRP tracing techniques

Summary Bidirectional transport of lectin conjugated horseradish peroxidase was employed to investigate the relative distribution qf the cerebellar and cortical connections of the nucleus of Darkschewitsch in the cat. Injection of horseradish peroxidase into the deep cerebellar nuclei produced terminal labeling which extended throughout the length of the contralateral nucleus of Darkschewitsch and into the perifascicular region. Injection of horseradish peroxidase into the pericruciate cortex produced both ipsilateral terminal labeling which extended throughout the length of the nucleus of Darkschewitsch and into the perifascicular region, and ipsilateral retrograde neuronal labeling. Labeled neurons displayed a variety of shapes and sizes, were more numerous in sections cut at rostral levels of the nucleus of Darkschewitsch, and were located both within and outside fields of terminal labeling. Comparison of the distribution of labeling following cerebellar and cortical injections indicates that convergence and overlap of input from these two sources occur in the nucleus of Darkschewitsch. These findings provide the morphological basis for integration of cerebellar and cortical information in this nucleus which may, in turn, influence output from neurons which project to the cortex or to the inferior olivary nucleus.Abbreviations ASG Anterior sigmoid gyrus - CA Cerebral aqueduct - CoS Coronal sulcus - CrS Cruciate sulcus - dCN Deep cerebellar nuclei - FR Fasciculus retroflexus - INC Interstitial nucleus of Cajal - ION Inferior olivary nucleus - IP _A Interposed anterior nucleus - IP _P Interposed posterior nucleus - L Lateral nucleus - LV Lateral vestibular nucleus - M Medial nucleus - ND Nucleus of Darkschewitsch - PAG Periaqueductal gray - PSG Posterior sigmoid gyrus - PSS Presylvian sulcus - V ₃ Third ventricle - y Cell group y A preliminary report of this study was presented at the Seventeenth Meeting of the Society for Neuroscience in New Orleans, LA, November 1987     

Brainstem organization of efferent projections to the guinea pig cochlea studied using the fluorescent tracers fast blue and diamidino yellow

Summary Intracochlear injection of the fluorescent retrograde neuronal tracers fast blue and diamidino yellow was used to investigate the distribution within the brainstem of neurones projecting to the cochlea in the guinea pig. The overall pattern of distribution of cells within the brainstem auditory nuclei was the same for both tracers and was also in broad agreement with recent studies in this species using horse-radish peroxidase as the neuronal tracer. However, the total number of neurones found (mean of 1234 projecting to each cochlea) was significantly greater than that reported using horseradish peroxidase, largely as a result of more small labelled neurones being detected within the lateral superior olivary nucleus ipsilateral to the injected cochlea. The yield of labelled cells was greatest in animals in which care was taken to perfuse the whole length of the cochlear epithelium. After bilateral injections of both tracers, double-labelled cells were found in small numbers within all the large neurone medial system nuclei and the ventral nucleus of the lateral lemniscus. It was concluded that between 1–5% of the medial system neurones project to both cochleas.