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Effects of excitotoxic lesions of the substantia innominata, ventral and dorsal globus pallidus on visual discrimination acquisition, performance and reversal in the rat.

Rats received infusions of ibotenic acid into the substantia innominata, in the region of the nucleus basalis of Meynert (nbM), before and after training on simple (simultaneous) and conditional visual discriminations. The ibotenate infusions reduced cortical choline acetyltransferase (ChAT) levels by about 20%, destroyed many ChAT-immunoreactive neurons in the nbM, but also caused the loss of many neurons in the substantia innominata and adjacent areas. These lesions did not impair the acquisition and performance of a simple visual discrimination, but did impair reversal of the discrimination and the performance of a conditional visual discrimination. However, the degree of impairment was unrelated to the degree of cortical ChAT loss. Ibotenic acid lesions to the dorsal globus pallidus also impaired reversal of discrimination but left acquisition and performance unaffected. Striatal dopamine depletion produced by 6-hydroxydopamine (6-OHDA) infusions into the mid-ventral caudate nucleus impaired performance of the simultaneous visual discrimination. Cortical noradrenaline depletion produced by 6-OHDA lesions of the dorsal noradrenergic bundle either alone or in combination with ibotenic acid lesions of the substantia innominata had no effect on acquisition of the discrimination. It is concluded that ibotenic acid lesions of the substantia innominata or to the dorsal globus pallidus affect learning and performance of conditional visual discrimination performance and impair reversal learning without affecting the capacity to discriminate visual events. These results are compared to those following cortical noradrenaline depletion or striatal dopamine loss.

Animals↗

Evidence for a projection from the lateral preoptic area and substantia innominata to the 'mesencephalic locomotor region' in the rat.

A series of anatomical and electrophysiological experiments have been carried out to examine the organization of a direct projection from the substantia innominata and the lateral preoptic area of the hypothalamus, referred to collectively as the subpallidal region, to the pedunculopontine nucleus and adjacent parts of the dorsal midbrain in the adult rat. In the first series of experiments, the retrogradely transported fluorescent tracer SITS, which does not appear to be taken up by fibers-of-passage, was injected into the pedunculopontine nucleus, and the distribution of labeled neurons was plotted in the substantia innominata and lateral preoptic area, as well as in adjacent regions including the medial preoptic area, the bed nucleus of the stria terminalis and parvocellular parts of the paraventricular nucleus. Then, the anterogradely transported lectin PHA-L, which also does not appear to be taken up in effective amounts by fibers-of-passage, was injected into parts of the substantia innominata and lateral preoptic area that project directly to the pedunculopontine nucleus. These experiments demonstrated that fibers from both regions descend through the medial forebrain bundle and give rise in the pedunculopontine nucleus to a terminal field that contains many structures with the appearance of terminal boutons. They also indicated that individual fibers from the subpallidal region innervate both the pedunculopontine nucleus and adjacent parts of the central gray, and that the pathway innervates areas along the length of the medial forebrain bundle on its way to the dorsal midbrain. In a third series of experiments the retrogradely transported fluorescent tracer True Blue was injected into upper thoracic levels of the spinal cord, and it was found that the region of the pedunculopontine nucleus that receives the densest input from the subpallidal region contained many retrogradely labeled neurons on both sides of the brain. And finally, a series of electrophysiological experiments demonstrated that single-pulse stimulation of the substantia innominata and the lateral preoptic area altered the firing rate of a majority of the neurons in and around the pedunculopontine nucleus, and that excitatory and inhibitory responses occurred about equally. These results clearly suggest that the subpallidal region projects directly to the pedunculopontine nucleus and adjacent regions including the central gray and the superior colliculus.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

Cortical projections of the nucleus of the diagonal band of Broca and of the substantia innominata in the rat: an anatomical study using the anterograde transport of a conjugate of wheat germ agglutinin and horseradish peroxidase.

The projections of the nucleus of the diagonal band of Broca and of the substantia innominata to the cerebral cortex were studied in the rat, using the anterograde transport of wheat germ agglutinin conjugated with horseradish peroxidase. Following diagonal band injections, fibers were observed ascending in the septum and reaching the cingulate cortex. They had a rostrocaudal, horizontal direction, mostly in layer VI and could be followed over long distances on sagittal sections. The fibers gave off collaterals which were seen ascending in the cerebral cortex and reaching more superficial layers. Following substantia innominata injections, fibers were observed to take two routes: the first one identical to the one described above and a second through the caudate-putamen reaching the temporo-insular cortex. Terminal fields had a more diffuse distribution following substantia innominata than following diagonal band injections. No clear laminar pattern of termination was observed. However the density of terminals was higher in layers IV, V and VI than in layers I, II and III. Since the conjugate used is not taken up by fibers of passage, this pattern of connection is believed to reflect the organization of the projection of the nucleus of the diagonal band and of the substantia innominata to the cerebral cortex.

Animals↗

Autoradiographic mapping of cerebral blood flow responses to cholinergic stimulation of the rat substantia innominata: modulatory effect of galanin.

In order to analyze the precise cerebrovascular effects of a specific cholinergic stimulation of the rat substantia innominata and their modulation by galanin, cerebral blood flow was measured by the [14C]-iodoantipyrine autoradiographic method in anesthetized (urethane and alpha-chloralose), artificially ventilated male Sprague-Dawley rats that received a microinjection into the substantia innominata of saline (n = 7), or 63 pmol of galanin (n = 8), or 50 nmoles of carbachol (n = 6) or a coinjection of carbachol and galanin (n = 8). Significant carbachol-induced cerebral blood flow increases were noted in ipsilateral cortices (+36%, p < 0.01 in the cingulate to +82%, p < 0.01 in the parietal somatosensory cortices), but also in ipsilateral hippocampus and ipsilateral thalamus. These cerebral blood flow increases were abolished by the coinjection of carbachol and galanin, while infusions of galanin alone failed to affect cerebral blood flow. Cholinergic stimulation of the substantia innominata represents thus a good model for the analysis of the detailed pharmacological properties of the cholinergic vasodilatatory basalocortical system. The existence of an inhibitory galaninergic modulation of this system could be of particular interest, in terms of cerebrovascular reactivity, in various neurodegenerative states.

Animals↗

Autoradiographic study of the cerebrovascular effects of stimulation of the substantia innominata: convenient stimulation paradigm.

The aim of this study was to determine the distribution within the whole brain of the vascular effects of stimulation of the substantia innominata. This basal forebrain nucleus is the major cholinergic input in the neocortex in the rodent. The local cerebral blood flow was measured by the autoradiographic [14C]iodoantipyrine technique in a group of control and a group of stimulated unanesthetized rats. The substantia innominata was electrically stimulated through a chronically implanted electrode. The stimulation induced blood flow increases exceeding 200% in the hemisphere ipsilateral to the stimulation and 100% in the contralateral hemisphere compared to the control group. The ipsilateral vasodilations were observed not only in the cortical areas but also in some subcortical structures. Comparison with previous data suggests that part of the effects is due to cholinergic neurons of the substantia innominata and part to non-cholinergic neurons and indirect effects. However, only two out of eight stimulated rats displayed this response. The low reproducibility of the results is discussed, considering the stimulation paradigm which has been developed for future measurements of the cerebral glucose utilization which requires a long duration stimulation period.

Animals↗

Third group of neostriatofugal neurons: neurokinin B-producing neurons that send axons predominantly to the substantia innominata.

Neostriatal neurons that produce neurokinin B were investigated immunocytochemically in the rat brain with an antibody against the C-terminal portion of the precursor prepropeptide of neurokinin B, preprotachykinin B (PPTB). PPTB-immunoreactive neurons were scattered throughout the neostriatum and constituted 5.1% of neostriatal neurons. They were immunopositive for projection neuron markers, such as precursor peptides of substance P, enkephalins, and dynorphins, but negative for intrinsic neuron markers, suggesting that PPTB was expressed in neostriatal projection neurons. However, PPTB-immunoreactive neurons were immunonegative for dopamine- and cyclic AMP-regulated phosphoprotein, which is known to be produced by striatopallidal and striatonigral neurons. Furthermore, almost no PPTB-immunoreactive axon terminals were observed in the substantia nigra or globus pallidus. The authors then made large kainic acid lesions in the neostriatum to reveal the target areas of PPTB-producing neurons and observed a decrease in PPTB-immunoreactive fibers in the sublenticular portion of the substantia innominata and, to much lesser extent, in the bed nucleus of the stria terminalis and central nucleus of the amygdala. After injection of wheat germ agglutinin into the substantia innominata, PPTB immunoreactivity was detected in many retrogradely labeled neostriatal neurons. In contrast, no PPTB immunoreactivity was observed in striatonigral or striatopallidal neurons after injection of retrograde tracers into the substantia nigra or globus pallidus. Thus, neurokinin B-producing neostriatal neurons were considered to send projection fibers predominantly to the substantia innominata. Furthermore, PPTB-immunoreactive axonal swellings were closely apposed to neurokinin B receptor-immunoreactive dendrites in the substantia innominata. Overall, the present results indicate that the rat brain possesses a chemically and hodologically unique neostriatofugal pathway in addition to the direct and indirect pathways.

Animals↗

The substantia innominata in Alzheimer's disease: an histochemical and biochemical study of cholinergic marker enzymes.

Acetylcholinesterase staining was examined in the substantia innominata of 3 normal human brains. Large intensely stained neurones were seen within the region of the basal nucleus of Meynert which is believed to be the origin of the cholinergic projection to the neocortex in animals. On the basis of the acetylcholinesterase staining pattern, the substantia innominata was dissected from post-mortem brain tissue of 19 cases of Alzheimer's disease (AD) and 16 controls so as to include the basal nucleus. Choline acetyltransferase (ChAT) activity was found to be reduced in the substantia innominata and amygdala in AD but not in the adjacent lentiform nucleus and hypothalamus.

Acetylcholinesterase↗

Neuronal responses related to the novelty and familarity of visual stimuli in the substantia innominata, diagonal band of Broca and periventricular region of the primate basal forebrain.

This study examined correlates of memory processes in neuronal activity recorded from the substantia innominata, the diagonal band of Broca and the periventricular region or the basal forebrain of monkeys performing recognition and/or visual discrimination tasks. Two types of neurons were found that responded differentially to stimuli on the basis of their novelty or their familiarity. Neurons (5/572) in the periventricular region rostral to the thalamus and caudal to the anterior commissure responded to familiar stimuli with increases in firing rate. Neurons in the substantia innominata (16/1058) and in the diagonal band (14/489) responded maximally to novel stimuli, with smaller responses to reappeared presentations of these stimuli. The properties of these two types of neurons were similar in three respects: (1) the magnitude of the differential response to novel and familiar presentations of stimuli were largest for stimuli presented on successive trials, and were attenuated for stimuli that had not been seen for some intervening trials; (2) both types of neurons responded to highly familiar stimuli as if they were novel if they had not been recently seen; (3) both types responded to two- and three-dimensional stimuli; and were recorded in rhesus monkeys trained on the recognition tasks and in untrained cynomolgus monkeys. An injection of HRP into the periventricular region of one monkey resulted in retrograde labeling of ventromedial regions of the prefrontal, cingulate and temporal cortices, of the amygdala, medial thalamus, supramammillary region of the midbrain. These data indicate that information about the novelty, familiarity or recency of presentation of visual stimuli is reflected in the responses of some basal forebrain neurons. Neurons in the substantia innominata and diagonal band of Broca could not be distinguished using the battery of applied tests, suggesting functional mechanisms common to both regions. The results of the anatomical experiment suggest that ventromedial limbic cortical and subcortical regions projecting to or through the periventricular region may be important for the transmission of information about visual stimuli and for memory function.

Action Potentials↗

[Neuronal reactions of the substantia innominata in the cat during an instrumental conditioned reflex].

Neuronal impulse activity in the substantia innominata has been studied in cats during conditional placing with food reinforcement. 44.3% of neurons changed their activity in trained cats. Activity of 24.3 neurons correlated with conditional sound stimuli. Reactions correlating with the conditional movement were weak. The most intensive and prolonging response was observed during food reinforcement. In some cases this reaction appeared 500 ms before the reinforcement. It is assumed that involvement of cholinergic neurons of the substantia innominata in reinforcement assisted trace phenomenon maintenance in neocortex neurons.

Acoustic Stimulation↗

Comparative effects of ibotenic acid- and quisqualic acid-induced lesions of the substantia innominata on attentional function in the rat: further implications for the role of the cholinergic neurons of the nucleus basalis in cognitive processes.

Two experiments examined the effects of excitotoxic lesions of the substantia innominata on cholinergic activity in the neocortex and on performance in a paradigm measuring selective attention in the rat. In Expt. 1, ibotenate-induced lesions produced approximately 30% reductions in cortical choline acetyltransferase (ChAT) activity, and damage to wide regions of the substantia innominata and ventral pallidum. The rats were impaired in their ability to localize brief visual targets in a serial reaction time task, as measured by reduced choice accuracy. This impairment was particularly evident at short stimulus durations, but the lesioned rats did not exhibit evidence of primary visual sensory dysfunction and exhibited only minor deficits when the stimuli were presented unpredictably. The deficit was exacerbated when distracting white noise was interpolated into the task. The rats with lesions were also slower to respond correctly, probably resulting partly from the adoption of a speed/error trade-off strategy, and were slower to collect earned food pellets, although they made no more errors of omission than controls. In Expt. 2, quisqualate-induced lesions produced fewer signs of non-specific damage and 50% reductions in cortical ChAT activity. This lesion produced generally qualitatively similar, but weaker effects to those of ibotenate-induced lesions. It was notable that many of the deficits following either ibotenate- or quisqualate-induced lesions lasted for several months after surgery. The results are discussed in terms of the cholinergic hypothesis of cognitive dysfunction. It is argued that lesions of the substantia innominata, including the magnocellular cholinergic neurons of the nucleus basalis of Meynert, produce deficits in attentional processing, which may not result from damage specifically to cholinergic cells. However, the longevity of the effects makes these preparations suitable for further exploration of the restorative effects of cholinergic treatments.

Animals↗

[Electrophysiological identification of neurons of the substantia innominata].

Recently in senile dementia of Alzheimer type, neuronal loss of cholinergic neurons in the substantia innominata is described. However these findings are based on morphological studies and, so far, these neurons have not been identified physiologically. In this study we tried to record these neurons electrophysiologically by antidromic activation. Cats were anesthetized with sodium pentobarbital and paralyzed with gallamine triethiodide. Three arreys of 3 or 4 silver-ball stimulating electrodes were fixed to the surface of the cerebral cortex. Each electrode was stimulated by rectangular pulses of 1 msec duration. Recording electrodes were glass micropipettes filled with 2 M NaCl saturated with fast green FCF and inserted into the ipsilateral substantia innominata stereotaxically. Twenty-eight neurons were constituted of mainly negative component; suggesting that they were recorded from cell bodies, not from axons. Also they responded in an all-or-none manner and showed constant latencies when stimulus intensities were at threshold level. When paired stimuli were applied, the latency of the action potential to the second stimulus was equal to that of the first one. These neurons were, therefore, considered to be activated antidromically. These neurons had axons of very slow conduction velocities. They are divided into two groups according to conduction velocities. The first group had mean conduction velocity of 2.3 m/sec and they responded to rather high frequency stimuli. The second group had mean conduction velocity of 1.0 m/sec and neurons belonging to this group showed quite long refractory periods. Based on conduction velocity analysis, the former is thought to include neurons with myelinated axons and the latter those with unmyelinated ones.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of lesions in the substantia innominata on active avoidance task and cerebral glucose metabolism.

Abnormalities in the central cholinergic system have been reported in Alzheimer disease. The present study was designed to evaluate the effects of the central cholinergic neurons in the substantia innominata on cognitive functions and cerebral metabolism in rats. Forty-nine Wistar rats were trained for an active avoidance task for 5 days. On Day 5, rats which had acquired the avoidance response were divided into two groups. The lesioned group (n = 7) received ibotenic acid in the substantia innominata bilaterally and the sham-operated group (n = 7) received the vehicle only. From Day 8 to 12, the rats were again trained for the avoidance task. On Day 12, the local cerebral metabolic rate for glucose was measured by the 14C-2-deoxyglucose method. The rats in the lesioned group revealed severe deficits in retention and re-acquisition of the active avoidance task. In addition, the glucose metabolism was decreased in the cerebral cortices, especially in the frontal cortex (p less than 0.05). The central cholinergic neurons of the substantia innominata may be closely related to cognitive functions and to glucose metabolism in the cortices.

Animals↗

High density of benzodiazepine binding sites in the substantia innominata of the rat.

In order to study the neuronal basis of the pharmacological interactions between benzodiazepine receptor ligands and cortical cholinergic turnover, we examined the regional distribution of specific benzodiazepine binding sites using in vitro autoradiography. In the basal forebrain, the substantia innominata contained a high density of [3H]lormetazepam (LMZ) binding sites (Bmax = 277 fmol/mg tissue; Kd = 0.55 nM). The label could be displaced by diazepam (IC50 = 100 nM), the benzodiazepine receptor antagonist beta-carboline ZK 93426 (45 nM) and the partial inverse agonist beta-carboline FG 7142 (540 nM). It is hypothesized that the amnesic effects of benzodiazepine receptor agonists are exerted through benzodiazepine receptors which are situated on cholinergic neurons in the substantia innominata and are involved in a tonic inhibition of cortical acetylcholine release. The benzodiazepine receptor antagonist ZK 93426 may exert its nootropic effects via benzodiazepine receptors in the substantia innominata and, consequently, by disinhibiting cortical acetylcholine release.

Animals↗

Responses of monkey prefrontal neurons during a visual tracking task reinforced by substantia innominata self-stimulation.

The role of the substantia innominata (SI) in the generation of a behavior mediated by the prefrontal (PF) cortex was examined in two Japanese monkeys. PF neuronal activities related to a visual tracking task by wrist movement were recorded and neural responses to SI stimulation were analyzed. Sixty-six neurons showed task-related activity and were classified into 3 types. Type 1 (n = 31) showed transient activation during the movement. Type 2 (n = 26) showed gradually increasing activity before the reward presentation. Type 3 (n = 9) showed tonic activation from the GO signal to the reward presentation. Antidromic and orthodromic responses to SI stimulation were observed in every type. In Type 1, the percentage of antidromically activated neurons (26%) was similar to that of orthodromically activated ones (16%), but in Type 2, 83% of responding neurons showed antidromic responses, and 56% of Type 3 showed orthodromic responses. These results show that the different types of PF neurons have different anatomical relations to SI. Although orthodromically activated neurons were fewer than antidromically activated neurons, many orthodromically activated neurons showed movement-related activity. This suggests that ICSS at SI facilitates the generation of the behavior through the afferent pathway to PF.

Animals↗

Substantia innominata projection to caudate nucleus in macaque monkeys.

The distribution of the cells of origin projecting from the substantia innominata to the caudate nucleus was studied in macaque monkeys by means of the retrograde axonal transport of horseradish peroxidase (HRP). The HRP was injected, after callosotomy, into the exposed head of the caudate nucleus. Labeled cells appeared in the ipsilateral substantia innominata, extending rostrally to the level of the optic chiasma.

Afferent Pathways↗

Biogenic amines and their metabolites in Alzheimer's disease: noradrenaline, 5-hydroxytryptamine and 5-hydroxyindole-3-acetic acid depleted in hippocampus but not in substantia innominata.

The following monoamine transmitters and related neurochemicals were analyzed in hippocampus and substantia innominata from postmortem brain tissue of subjects with senile dementia of the Alzheimer's type (SDAT): tryptophan (TP), 5-hydroxytryptamine (5-HT), 5-hydroxyindole-3-acetic acid (5-HIAA), noradrenaline (NA), dopamine (DA) and homovanillic acid (HVA). The results were compared to those obtained in postmortem brain samples obtained from normal subjects matched for age, sex and time between collection of tissue and freezing. There were no significant differences in concentrations of TP, DA or HVA between normals and SDAT subjects in either brain region. While hippocampal 5-HT, 5-HIAA and NA concentrations were markedly reduced in SDAT patients relative to normals, the substantia innominata concentrations of these three were not significantly different between controls and SDAT patients, emphasizing the importance of detailed regional analyses when conducting neurochemical studies in SDAT.

Aged↗

Efferent connections of the substantia innominata in the rat.

The efferent connections of the substantia innominata (SI) were investigated employing the anterograde axonal transport of Phaseolus vulgaris leucoagglutinin (PHA-L) and the retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). The projections of the SI largely reciprocate the afferent connections described by Grove (J. Comp. Neurol. 277:315-346, '88) and thus further distinguish a dorsal and a ventral division in the SI. Efferents from both the dorsal and ventral divisions of the SI descend as far caudal as the ventral tegmental area, substantia nigra, and peripeduncular area, but projections to pontine and medullary structures appear to originate mainly from the dorsal SI. Within the amygdala and hypothalamus, which receive widespread innervation from the SI, the dorsal SI projects preferentially to the lateral part of the bed nucleus of the stria terminalis; the lateral, basolateral, and central nuclei of the amygdala; the lateral preoptic area; paraventricular nucleus of the hypothalamus; and certain parts of the lateral hypothalamus, prominently including the perifornical and caudolateral zones described previously. The ventral SI projects more heavily to the medial part of the bed nucleus of the stria terminalis; the anterior amygdaloid area; a ventromedial amygdaloid region that includes but is not limited to the medial nucleus; the lateral and medial preoptic areas; and the anterior hypothalamus. Modest projections reach the lateral hypothalamus, with at least a slight preference for the medial part of the region, and the ventromedial and arcuate hypothalamic nuclei. Both SI divisions appear to innervate the dorsomedial and posterior hypothalamus and the supramammillary region. In the thalamus, the subparafascicular, gustatory, and midline nuclei receive a light innervation from the SI, which projects more densely to the medial part of the mediodorsal nucleus and the reticular nucleus. Cortical efferents from at least the midrostrocaudal part of the SI are distributed primarily in piriform, infralimbic, prelimbic, anterior cingulate, granular and agranular insular, perirhinal, and entorhinal cortices as well as in the main and accessory olfactory bulbs. The cells of origin for many projections arising from the SI were identified as cholinergic or noncholinergic by combining the retrograde transport of WGA-HRP with histochemical and immunohistochemical procedures to demonstrate acetylcholinesterase activity or choline acetyltransferase immunoreactivity. Most of the descending efferents of the SI appear to arise primarily or exclusively from noncholinergic cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Efferent connections of the striatopallidal and amygdaloid components of the substantia innominata in the cat: projections to the nucleus accumbens and caudate nucleus.

Enkephalin immunoreactivity is used to divide the feline substantia innominata into circumscript subregions, i.e. the "striatopallidal system" and the "extended amygdala". In addition, enkephalin immunoreactivity is used to subdivide the striatopallidal system into two distinct areas, i.e. the subcommissural part of the globus pallidus displaying high enkephalin immunoreactivity and the ventral pallidum displaying moderate enkephalin immunoreactivity. The anterograde axonal transport of Phaseolus vulgaris-leucoagglutinin is used to study the efferents of these areas innervating the caudate nucleus and the nucleus accumbens. It is found that the enkephalin-immunoreactive subcommissural part of the globus pallidus as well as the dorsal enkephalin-immunoreactive regions of the extended amygdala project topographically along a rostrocaudal and mediolateral dimension to the nucleus accumbens. The far rostral parts of the caudate nucleus are found to be innervated by the subcommissural part of the globus pallidus whereas the extended amygdala has no such connection. This pathway is also found to be topographically organized along a mediolateral dimension. The non-enkephalin-immunoreactive area ventral and lateral to the subcommissural part of the globus pallidus is found to have no projections to the nucleus accumbens and caudate nucleus. This region rather innervates the olfactory tubercle. In contrast to the striatopallidal system the sublenticular part of the extended amygdala preferentially projects to the adjoining part of the extended amygdala, i.e. the bed nucleus of the stria terminalis. However, the ventral regions preferentially innervate the medial division of the bed nucleus of the stria terminalis whereas the dorsal regions preferentially innervate the lateral division of the bed nucleus of the stria terminalis. These data indicate that the differential forebrain systems represented in the feline substantia innominata, i.e. the striatopallidal system and extended amygdala have differential output stations. The results are discussed in view of the role of the subcommissural part of the globus pallidus and the nucleus accumbens in orofacial dyskinesia and schizophrenia, respectively.

Amygdala↗