Supraoptic nucleus afferents from the main olfactory bulb--I. Anatomical evidence from anterograde and retrograde tracers in rat

The morphological features of a putative connection between the main olfactory bulb and the supraoptic nucleus of the rat was studied using a combination of anatomical techniques. Immunocytochemistry of neurophysin-containing processes were employed to delineate morphological features of supraoptic dendrites. Main olfactory bulb efferents to the supraoptic nucleus were studied by injection of the anterogradely transported substances, wheatgerm agglutinin conjugated horseradish peroxidase or Phaseolus vulgaris leucoagglutinin, into the main olfactory bulb. To confirm the results of these studies, the distribution of retrogradely labeled cells within the main olfactory bulb was determined after injection of rhodamine-labeled latex microspheres or Fluoro-Gold into the supraoptic nucleus. Neurophysin immunocytochemistry revealed the supraoptic nucleus dendritic plexus which coursed anteroposteriorly beneath supraoptic somata. Additionally, a portion of this plexus also projected ventrolaterally into periamygdaloid areas, a feature of supraoptic architecture which is not generally appreciated. The anterograde tracers labeled main olfactory bulb efferents including a dense plexus of terminals and fibers ventrolateral to the ipsilateral supraoptic nucleus. The pattern of anterogradely labeled fibers and terminals appeared to overlap with the distribution of ventrolaterally projecting neurophysin-containing processes. Since the latter consists of dendritic processes of supraoptic origin, this suggests that the main olfactory bulb projects to the supraoptic nucleus. Injections of rhodamine-labeled latex microspheres or Fluoro-Gold resulted in retrogradely labeled mitral cells throughout the ipsilateral main olfactory bulb. Taken together, these anatomical studies demonstrate a direct projection from the main olfactory bulb to the supraoptic nucleus of the rat. A comparison electrophysiological study confirmed these results.     

Cerebellar afferents from the lateral reticular nucleus in the rat

The projection from the lateral reticular nucleus to the cerebellum was examined using the retrograde fluorescent double labelling method in the rat. A quantitative analysis of the extent of axonal branching and of nucleotopic (topographic) organization in this projection was carried out. It was shown that lateral reticular nucleus neurons show axonal branching both within and between parasagittal zones of the cerebellar cortex, but that branching within zones is more common. Neurons projecting to different zones were found to be significantly spatially separated, but those projecting to different sites within the same zone were intermingled. Thus a limited degree of nucleotopic organization was found. The results are discussed in relation to the parasagittal zonation in the olivocerebellar projection.     

Cerebellar afferents from the nucleus of the solitary tract.

The cerebellar afferents from the nucleus of the solitary tract were studied in cat by means of retrograde axonal transport of horseradish peroxidase. Labelled cells occurred bilaterally in the nucleus of the solitary tract following injections in various folia of the cerebellar vermis and in the flocculus (the positive cases are shown in Fig. 1A). Injections in the anterior lobe vermis labelled cells in the caudal part of the nucleus, injections in the posterior vermis labelled cells in the rostral part (Fig. 2). The findings are discussed in relation to other efferent and afferent connections of the nucleus of the solitary tract.     

Neuronal responses of the anterior commissural nucleus to osmotic stimulation and angiotensin II in hypothalamic slices in the rat.

The firing rate and pattern of activity of neurons in the anterior commissural nucleus (ACN), which was rich in oxytocin-containing neurons, were studied electrophysiologically in hypothalamic slices. Extracellular recording showed that most ACN neurons exhibited irregular or regular continuous spontaneous unit activity. Other neurons showed short burst patterns of activity or were silent. The majority of ACN neurons were activated by bath application of angiotensin II, and a substantial number of them showed inhibitory or excitatory responses to hypertonic bathing medium. These results indicate that magnocellular neurosecretory neurons in the ACN may participate in the regulation of water balance.     

Olfactory bulb networks revealed by lateral olfactory tract stimulation in the in vitro isolated guinea-pig brain

Olfactory information processing is mediated by synaptic connections between the olfactory bulbs (OBs) and piriform-limbic cortices. Limited accessibility using common in vivo and in vitro preparations has hindered previous attempts to define these synaptic interactions. We utilized the isolated guinea-pig brain preparation to overcome these experimental limitations. Previous studies demonstrated extensive functional preservation in this preparation maintained in vitro by arterial perfusion. Field potential laminar profiles were performed with multi-channel probes in the OB following stimulation of both the lateral olfactory tract (LOT) and the anterior piriform cortex (APC). Current-source density analysis was carried out on laminar profiles to reconstruct current sinks/sources associated with intrinsic synaptic activities. LOT stimulation induced sequentially i) an antidromic population spike (at 2.66+/-0.39 ms) located in the mitral cell layer that was resistant to 100 Hz high-frequency stimulation (HFS) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (10 microM), ii) a component located in the external plexiform layer at 3.85+/-0.63 ms that was unaffected by HFS, iii) a large amplitude potential (peak amplitude at 5.84+/-0.58 ms) generated in the external plexiform layer, abolished by HFS and CNQX, but not by bicuculline (50 microM), iv) a late response (onset at 20.00+/-2.94 ms) abolished by CNQX and enhanced by bicuculline. Stimulation of the APC also induced a late potential abolished by HFS and CNQX. Both APC-evoked and late LOT-evoked responses were abolished by a transverse cut to separate OB from APC. These results demonstrate in an isolated mammalian brain preparation the presence of reciprocal synaptic interactions between the OB and piriform cortical structures.     

Lack of centripetal connection between the lateral olfactory tract and the caudate nucleus. Evoked potential study in the rat

Summary In rats under alpha-chloralose or without general anesthesia the effect of a lateral olfactory tract stimulus on the caudate nucleus electric activity was explored by bipolar macroelectrodes. The stimulation effectiveness and the reactivity of recording sites to somatic stimuli were systematically tested. Under these conditions, no functional connection from the lateral olfactory tract to the caudate nucleus was demonstrated.     

Periaqueductal gray spike trains recorded in frontal or horizontal mesencephalic brain slices from the rat

Spontaneous spike trains were recorded from the periaqueductal gray matter (PAG) in frontal and horizontal mesencephalic slices in order to compare spontaneous activity of the slice preparation to previous in vivo records. The firing rates resembled those recorded in vivo. They were low notwithstanding the fact that the slicing procedure removed tonic inhibitory input to the PAG. The firing rates increased caudo-rostrally, a fact that had not been reported in vivo, and were lower in frontal sections. Several different spike trains were found, described and classified according to their temporal firing patterns. The spike sequences were usually simple stochastic processes, distributed as a Gaussian function with or without stochastic deletion of spikes.     

Evoked responses of pyramidal tract neurons to stimulation of the lateral hypothalamus

The evoked responses of identified pyramidal tract neurons of the pericruciate region of the cortex arising on stimulation of the posterior, tuberal, and anterior sections of the lateral hypothalamus were investigated in acute experiments on cats anesthetized with chloralose. The pyramidal tract neurons recorded were situated at a depth of 0.75–2.5 mm and on stimulation of the hypothalamus discharged with a latent period of 1.6–52.5 msec. They discharged most efficiently in the case of stimulation of the caudal half of the lateral hypothalamus. The pyramidal tract neurons activated by the hypothalamus were also tested by electrocutaneous stimulation of the four limbs. Of the neurons 75% responded to stimulation of several limbs, i.e., they had a broad bilateral receptive field, while 15% reacted to stimulation of one contralateral limb (either the anterior limb or the posterior limb), i.e., they had a small contralateral receptive field. Comparison of the latent periods of the anti- and orthodromic responses of the pyramidal tract neurons did not reveal any relationship between the magnitude of the latent period of the orthodromic response (on hypothalamic or electrocutaneous stimulation) and the type of pyramidal tract neuron (according to the axonal transmission velocity). No relationship between the latent period of hypothalamic stimulation and the magnitude of the latent period of the response to a peripheral stimulus was detected either.Translated from Fiziologicheskii Zhurnal SSSR, imeni I. M. Sechenova, Vol. 65, No. 5, pp. 661–667, May, 1979.     

Gastric afferents to the paraventricular nucleus in the rat

Summary Extracellular recordings were made from vasopressin (AVP) and oxytocin (OXT)-secreting cells in the paraventricular nucleus (PVN) of the hypothalamus in rats anesthetized with urethane-chloralose to determine the effects of electrical stimulation of vagal gastric nerves and gastric distension on their activity. Electrical stimulation of gastric branches of the vagus nerves inhibited 5 and excited 10 of 32 phasically firing neurosecretory cells. Approximately one third of the phasically firing neuro-secretory cells (9 out of 29 cells) were transiently inhibited by gastric distension; an effect which was completely abolished by bilateral cervical vagotomy. In contrast, gastric nerve stimulation excited 45 of 72 non-phasically firing paraventricular cells. Thirteen of 77 non-phasically firing cells tested were excited by gastric distension. We conclude that there are some sensory afferent inputs originating from gastric receptors and transmitted by gastric vagal afferents which inhibit the activity of AVP- secreting neurons in the PVN although other inputs excite the cells. Similar inputs also excite some of the putative OXT-secreting neurons in the PVN.     

Selective neuroinhibitory effects of taurine in slices of rat main olfactory bulb

Taurine is abundant in the main olfactory bulb, exceeding glutamate and GABA in concentration. In whole-cell patch-clamp recordings in rat olfactory bulb slices, taurine inhibited principal neurons, mitral and tufted cells. In these cells, taurine decreased the input resistance and caused a shift of the membrane potential toward the chloride equilibrium potential. The taurine actions were sustained under the blockade of transmitter release and were reversible and dose-dependent. At a concentration of 5 mM, typically used in this study, taurine showed 90% of its maximal effect. GABA(A) antagonists, bicuculline and picrotoxin, blocked the taurine actions, whereas the glycine receptor antagonist strychnine and GABA(B) antagonists, CGP 55845A and CGP 35348, were ineffective. These findings are consistent with taurine directly activating GABA(A) receptors and inducing chloride conductance. Taurine had no effect on periglomerular and granule interneurons. The subunit composition of GABA(A) receptors in these cells, differing from those in mitral and tufted cells, may account for taurine insensitivity of the interneurons. Taurine suppressed olfactory nerve-evoked monosynaptic responses of mitral and tufted cells while chloride conductance was blocked. This action was mimicked by the GABA(B) agonist baclofen and abolished by CGP 55845A; CGP 35348, which primarily blocks postsynaptic GABA(B) receptors, was ineffective. The taurine effect most likely was due to GABA(B) receptor-mediated inhibition of presynaptic glutamate release. Neither taurine nor baclofen affected responses of periglomerular cells. The lack of a baclofen effect implies that functional GABA(B) receptors are absent from olfactory nerve terminals that contact periglomerular cells. These results indicate that taurine decreases the excitability of mitral and tufted cells and their responses to olfactory nerve stimulation without influencing periglomerular and granule cells. Selective effects of taurine in the olfactory bulb may represent a physiologic mechanism that is involved in the inhibitory shaping of the activation pattern of principal neurons.     

Supraoptic neurosecretory cells: Synaptic inputs from the nucleus accumbens in the rat

Summary Synaptic inputs from the nucleus accumbens (ACB) to neurosecretory cells of the supraoptic nucleus (SON) were studied in the rat. One hundred and twenty SON neurones responded antidromically to pituitary stalk stimulation and were identified as neurosecretory cells. Sixty-three of these cells were identified as vasopressin-secreting cells and 45 as oxytocin-secreting cells by their spontaneous firing patterns. About one half of the vasopressin-cells and two thirds of the oxytocin-cells were responsive to stimulation of the basal forebrain including the ACB. More vasopressin-cells were excited than were inhibited, and oxytocin-cells were mainly inhibited. Depth profile of effective stimulation sites in the basal forebrain revealed that ACB stimulation selectively produced the responses. Most of those SON neurones responsive to ACB stimulation also responded to septal stimulation. A positive correlation was observed between responses to ACB and septal stimulation in each unit. After septal lesion, the number of SON neurones which were responsive to ACB stimulation was significantly decreased. In two rats, a single SON unit was tested for ACB stimulation both before and after septal lesion, and the previously observed synaptic inputs were not seen after the lesion. Fifty septal neurones projecting to the area including the SON were antidromically identified after SON stimulation. About one half of these neurones were excited by ACB stimulation. These results demonstrate the existence of a neural pathway from the ACB to the SON and suggest that the pathway is mediated by septal neurones.Supported by grant no. 57770111 from the Ministry of Education, Science and Culture, Japan     

Glutamergic transmission of neuronal responses to carbachol in rat dorsal cochlear nucleus slices

This study found that glutamate receptor antagonists block the excitatory effects of carbachol, a cholinergic agonist, on bursting neurons in the dorsal cochlear nucleus of rat brain slices. Among antagonists for glutamate receptor subtypes, those for non-N-methyl-D-aspartate ionotropic glutamate receptors were more potent than those for N-methyl-D-aspartate receptors. The glutamate receptor antagonists did not block the effects of carbachol on regularly firing neurons in the dorsal cochlear nucleus of the same slices. Antagonists for GABA or glycine receptors did not alter the effects of carbachol on bursting neurons. Effects of carbachol on bursting activity could be mimicked by application of glutamate or its agonist, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate, whose effects were not blocked by synaptic blockade. During carbachol application, increased release of glutamate and glycine from the dorsal cochlear nucleus part of brain slices was measured using high-performance liquid chromatography. Release of other amino acids showed no significant change. The results suggest that, in rat dorsal cochlear nucleus, cholinergic effects on regular and bursting spontaneous firing occur through different mechanisms. Cholinergic effects on regular neurons (which include fusiform cells) are direct, through muscarinic receptors. Cholinergic effects on bursting neurons (which include cartwheel cells) are indirect and involve glutamatergic neurotransmission, mostly via non-N-methyl-D-aspartate ionotropic receptors. The granule cell-parallel fiber pathway may be involved in this glutamatergic transmission.     

Anterior olfactory nucleus or lateral olfactory tract destruction in rats and changes in appetitive and aversive behavior

Previous research with rats demonstrated a continuum of behavioral and biochemical effects following variable olfactory system damage. In general, increased removal of olfactory tissue along an antero-posterior axis amplified both behavioral and biochemical changes. The present experiments were undertaken to explore possible roles of the anterior olfactory nucleus (AON) and lateral olfactory tract (LOT) in the observed behavioral phenomena. Destruction of either structure decreased passive avoidance performance but selectively affected shuttle box avoidance performance; viz, AON destruction decreased it slightly whereas LOT destruction enhanced it. Animals with LOT destruction were also more emotional than AONs when handled and, when extensively lesioned, were more active in an open field than all other groups. In a bar pressing task for continuous reinforcement, AONs pressed significantly more than SHAMs during extinction although the LOT mean was also higher. When lesion size was correlated with the scores on several tests, AONs correlated positively with passive avoidance, shuttle box avoidance, and time to reach the extinction criterion in the bar pressing test while LOT lesions correlated negatively with shuttle box avoidance and bar press extinction time. The results were discussed against a background of other olfactory system lesions and some possible roles of the LOT and AON in reactivity to stimulation and responsiveness to reinforcement were considered. The close anatomical connections between the olfactory and limbic systems and similar behavioral changes after both limbic and olfactory lesions suggested a possible basis for the observed effects.     

Responses of rat lateral hypothalamic neuronal activity to fastigial nucleus stimulation

1. The aim of this study was investigation of neuronal mechanisms underlying inputs from the fastigial nucleus (FN) to the lateral hypothalamic area (LHA). 2. In male anesthetized rats, 295 extracellular and 82 intracellular recordings of LHA responses to electrical stimulation of the FN, which elicited stimulus-locked pressor responses, were examined. 3. Contralateral FN stimulation evoked three types of responses in 48% of spontaneously firing LHA neurons: inhibition with 11 +/- 6 (SD) ms latency followed by excitation (30%), excitation with 15 +/- 12.5 ms latency (14%), and excitation followed by inhibition with 6 +/- 4 ms latency (4%). 4. Contralateral FN stimulation after transection of the inferior cerebellar peduncle (ICP), which resulted in a substantial fall of the fastigial pressor response, also evoked the three types of responses. These responses were unaffected by transection of the ICP. 5. Neuronal activity was recorded intracellularly from 82 LHA neurons, of which 36 (44%) responded to FN stimulation. Of the 36 neurons, 24 showed inhibitory postsynaptic potentials (IPSPs) with a mean latency of 7.5 +/- 2 ms. Of the 24 neurons, 16 were checked for change in IPSP latency with stimulus intensity, and 11 were considered to be monosynaptically connected since their latencies were constant when FN stimulation intensity was changed. The remaining 12 exhibited excitatory postsynaptic potentials (EPSPs) with a longer latency of 10.5 +/- 3 ms, which indicated polysynaptic conduction. The reversal potentials of the IPSP and EPSP were estimated to be about -77 mV and -13 mV, respectively. 6. Most glucose-sensitive neurons (78%), which were identified by their inhibition in response to electrophoretically applied glucose, were inhibited by FN stimulation, whereas only 7% of the glucose-insensitive neurons responded to such stimulation. 7. From the results, it was concluded that LHA neurons receive inhibitory monosynaptic and excitatory polysynaptic inputs from the FN via the superior cerebellar peduncle. These connections may contribute to hypothalamic modulation of feeding behavior.     

Suprarhinal cortex response to electrical stimulation of the lateral amygdala nucleus in the rat

Summary Electrical stimulation of the lateral amygdala nucleus was found to evoke field potentials and influence unitary activity in the suprarhinal cortex of anesthetized rats. Laminar distributions of the field responses consisted of positive waves in superficial layers, that reversed to electronegatives from a depth of 0.4–0.5 mm. This response was followed by a shallow electropositive wave deeper than 0.7–0.8 mm. Extracellularly recorded units were studied in the posterior agranular insular area of the suprarhinal cortex. The data revealed that stimulation of the lateral amygdala produced a train of small amplitude spikes in association with a negative slow potential. Furthermore, such stimulation invariably elicited an inhibition of the spontaneous firing of large amplitude spikes, in association with a positive slow potential. The onset of this inhibitory response always occurred at longer latency than the excitatory one. The small amplitude spikes may well represent the firing of inhibitory interneurons after lateral amygdala stimulation. The study suggests that a feedforward system of inhibition appears to be present in the connection between lateral amygdala and posterior agranular insular area of the suprarhinal cortex.     

Somatotopic termination of spinal afferents to the feline lateral cervical nucleus

Summary The labelling pattern of the feline lateral cervical nucleus (LCN) was investigated after pressure injections of lectin-conjugated horseradish peroxidase into cervical, thoracic or lumbar segments of the spinal cord. Sixteen cats received a 5–8 nl injection staining large parts of mainly the ipsilateral grey matter of a single segment. Light microscopic examination of frozen sections reacted with tetramethylbenzidine showed a somatotopic organization in the LCN. Rostral segments of the spinal cord projected mainly to rostroventral and medial parts of the ipsilateral LCN, while more caudally located segments projected to more dorsocaudal and lateral parts of the nucleus. Minor contralateral labelling with a similar somatotopic arrangement was seen in animals given cervical and lumbar injections. No significant labelling was found in the LCN of three control animals, the segmental injections of which were engaged mainly into the ipsilateral dorsal columns and the dorsolateral funiculus. Ultrastructural analysis in two animals which received multiple cervical or lumbar injections showed that about 70% of the peroxidase-positive structures in the LCN were boutons and the rest small myelinated axons. The precise termination pattern of ascending afferents to the LCN is compatible with the somatotopic organization of the other relay centres in the spino-cervicothalamic pathway.     

Descending influences on the responses of spinocervical tract neurones to chemical stimulation of fine muscle afferents

1. In cats, extracellular micro-electrode recordings were made from axons of the spinocervical tract (s.c.t.) in both the decerebrate state and during cold block of the spinal cord (reversible spinal state) to examine the effects of intra-arterial injection of algesic agents (bradykinin, potassium, 5-hydroxytryptamine) into the gastrocnemius-soleus (g.s.) muscle on the discharge behaviour of s.c.t. neurones.2. In the decerebrate state without cooling the spinal cord 13% of the cells (eleven out of eighty-three) responded to intra-arterial injection of bradykinin, 33% (twenty-two out of sixty-nine) to 5-hydroxytryptamine, and 38% (thirty-five out of ninety-one) to potassium injection.3. The general time course and the latency of the responses of s.c.t. cells induced by injection of pain-producing substances into the g.s. muscle reflect in many respects the activations of g.s. group III and group IV primary afferent units studied previously.4. For twenty-seven s.c.t. neurones the period of recording was long enough to record the responses of the same cell to injections of algesic agents in both the decerebrate and the reversible spinal state. In the reversible spinal state 83% (nineteen out of twenty-three) of the s.c.t. neurones tested with all the three substances responded to at least one of the algesic agents. In the decerebrate state the percentage was lower (39%).5. Reversible spinalization led not only to a significant increase in the number of s.c.t. neurones responding to the algesic agents used but also to an increase in the magnitude of the chemically induced responses.6. The mean latency of the responses of neurones that were activated in both preparations were shorter in the reversible spinal state than in the decerebrate state.7. Control experiments showed that the responses to bradykinin and potassium were entirely due to the nervous outflow from the g.s. muscle. In contrast, intra-arterially applied 5-hydroxytryptamine influenced the s.c.t. cells via unknown additional sites of action.8. The results indicate that muscular group III and/or group IV units excitable by algesic substances do project on to neurones of the spinocervical tract. Furthermore it is concluded that the responses of s.c.t. neurones to activation of fine muscle afferents by algesic agents are subject to a descending control similar to the well known descending modulation of their responsiveness to cutaneous input. Therefore, in addition to serving as a cutaneous pathway the spinocervical tract may take part in muscular nociception.