The density of pyramidal and nonpyramidal neurons in anterior cingulate cortex of schizophrenic and bipolar subjects.

BACKGROUND: A recent study reported a decreased density of nonpyramidal neurons (NPs) in layer II of the anterior cingulate (ACCx) and prefrontal (PFCx) cortices of schizophrenic brain that was most pronounced in schizoaffective subjects. Our study assessed whether a decrease of NPs in ACCx may show a stronger covariation with affective disorder. A cohort consisting of 12 normal control (CONs), 11 schizophrenic, and 10 bipolar subjects matched for age and postmortem interval (PMI) has been analyzed. METHODS: A two-dimensional technique was employed for counting cells in a large x,y sampling column that extended across layers I through VI of ACCx. RESULTS: There was a 27% reduction in the density of NPs in layer II of the bipolar group, whereas in the schizophrenic group, this density was 16.2% lower. There were no differences in NPs in layers III through VI of either the schizophrenic or bipolar group. Both groups also showed modest decreases of PNs in the deeper laminae; however, these differences were only significant in layer IV of the schizophrenic subjects. The density of glial cells was similar across the control, schizophrenic, and bipolar groups. An Abercrombie correction for cell size did not alter the nature of the results. Subjects both with and without neuroleptic exposure showed a lower density of NPs in layer II of bipolar subjects or PNS in deeper laminae of schizophrenic subjects. CONCLUSIONS: Overall, the findings reported here suggest that local circuit cells in layer II of ACCx may be decreased in bipolar disorder, whereas projection neurons in deeper laminae are decreased in schizophrenia.

A reduction of nonpyramidal cells in sector CA2 of schizophrenics and manic depressives.

BACKGROUND: Recent studies have suggested that there may be a preferential decrease of "nonpyramidal" neurons (NPs) in several corticolimbic regions of schizophrenic (SZ) brain. The current study was undertaken to determine whether a change in the density of pyramidal neurons (PNs) and NPs might be present in the hippocampal formation (HIPP) of SZ brain. METHODS: A spatial counting approach in which the location of each and every PN and NP in the stratum pyramidale of sectors CA1-4 was applied to 11 normal control (CONs) and 10 SZs matched for age and postmortem interval, as well as 4 manic depressive (MD) subjects matched for age. RESULTS: The data indicate that the CONs had approximately 10-20 times as many PNs than NPs in the various HIPP subfields. When the CON data were compared to those for the SZs, both the total number and density of PNs were found to be similar in all four sectors, while NPs were found to be selectively reduced by approximately 40% in CA2 of the SZ group. When the data were broken down according to patients with and without neuroleptic exposure, drug-free SZs showed a significant reduction in the density of NPs in CA2. The 4 MD cases both with and without neuroleptic exposure also showed a similar reduction of NPs in sector CA2. CONCLUSIONS: Taken together, the results of this study suggest that there may be a highly selective decrease in the number of NPs in sector CA2 that could play a contributory role in the pathophysiology of the major psychoses.

Uncoupling of GABA(A) and benzodiazepine receptor binding activity in the hippocampal formation of schizophrenic brain.

A recent postmortem study has reported that there is a widespread upregulation of GABA(A) receptor binding activity throughout most subregions of the hippocampal formation of schizophrenic brain. The current study has been undertaken to determine whether the benzodiazepine (BZ) receptor, which is a component of the GABA(A) receptor complex, may also be upregulated in schizophrenics. Using a low-resolution film autoradiographic technique to localize [3H]flunitrazepam binding, the subregional and laminar distribution of specific BZ receptor binding was found to parallel that of the GABA(A) site, except in the area dentata where BZ binding was approximately 73% higher in the outer molecular layer. When BZ receptor binding was compared in the same normal control (n = 15) and schizophrenic (n = 8) cases in which the GABA(A) receptor was analyzed, there were very few differences noted between the two groups, except for small, though significant, increases in the stratum oriens of CA3 (30%), the subiculum (20-30%) and the presubiculum (15-20%) of the patient group. These latter increases overlapped with the subregions and laminae in schizophrenics showing the most marked increases of GABA(A) receptor binding. Using a high-resolution technique to evaluate specific BZ receptor binding on different neuronal subtypes, no difference was observed on either pyramidal or nonpyramidal neurons of sector CA3 where GABA(A) receptor activity had been found to be significantly increased on the latter neuronal subtype. The potential confounding effects of age, postmortem interval and exposure to either benzodiazepine or neuroleptic drugs do not account for the lack of marked differences in BZ receptor binding in the schizophrenic group. Taken together, the results of this study are consistent with the possibility that defective GABAergic integration in schizophrenia may be associated with an uncoupling in the regulation of the GABA(A) and BZ receptors.

Up-regulation of GABAA receptor binding on neurons of the prefrontal cortex in schizophrenic subjects.

Recent investigations have reported a reduced density of interneurons and an increase of GABAA receptor binding occurring preferentially in layer II of the anterior cingulate cortex of schizophrenic subjects [Benes F.M. et al. (1992) J. Neurosci. 12, 924-929]. Since a reduction in the density of interneurons has also been found in layer II of the prefrontal cortex, this study has sought to determine whether an un-regulation of the GABAA receptor binding activity might also be found in this region of schizophrenics. A high-resolution autoradiographic analysis of bicuculline-sensitive [3H]muscimol (GABAA) receptor binding on individual neuron cell bodies in layers II, III, IV and VI has been applied to Brodmann area 10 from normal controls (n = 16) and schizophrenic (n = 7) subjects. A computer-assisted technique has been used under strictly blind conditions to determine whether differences in binding occur in the schizophrenic group. A significant increase of GABAA receptor binding activity has been observed in layers II, III, V and VI in the schizophrenic group. When the binding is expressed as a density with respect to neuronal cell size, there is a gradient of binding across layers II, III, V and VI, with neuronal cell bodies in layer II having the greatest density of grains. When different subpopulations of neurons distinguished according to size criteria are examined separately, large (pyramidal) neurons show significantly higher binding, particularly in layer II, where it was increased by 90% in schizophrenics. Small (non-pyramidal) cells do not show significant differences in binding in schizophrenics, except in layer VI, where there was a 135% increase. Potential confounding effects from age and post mortem interval do not explain the differences between the two groups, because both young and old schizophrenics, as well as schizophrenics with long and short post mortem intervals, showed increased GABAA receptor binding activity when compared to control cases distinguished in a corresponding fashion. These data suggest that there may be a preferential reduction of inhibitory GABAergic inputs to pyramidal neurons, particularly in layer II of the preferential cortex, in schizophrenia. This change could potentially result in an increased excitatory outflow from the prefrontal area to other cortical regions of the schizophrenic brain. Overall, these results are consistent with the idea that reduced amounts of GABAergic activity in the prefrontal cortex could be related to a perinatal disturbance and could be a potentially important component of the pathophysiology of psychosis.

Increased interaction of dopamine-immunoreactive varicosities with GABA neurons of rat medial prefrontal cortex occurs during the postweanling period.

A double immunofluorescence technique has been used to assess postnatal maturational changes in the extent to which dopamine-immunoreactive (DA-IR) varicose fibers form contacts with gamma-aminobutyric acid (GABA)-immunoreactive (IR) neuronal cell bodies in rat medial prefrontal cortex (mPFCx). Two separate measures of interaction, the percentage of GABA-IR cell bodies having DA-IR varicosities in apposition and the number of such profiles in contact with any given GABA-IR cell, were assessed. Between birth and adulthood, there was a progressive linear increase (r = 0.75, P < or = 0.0005) in the percentage of GABA-IR cell bodies having at least one DA-IR varicosity in apposition. While the number of varicosities in contact with any given GABA cell body showed very little change during the preweanling period, later during the postweanling interval, this parameter increased in a curvilinear fashion toward adult levels (r = 0.81, P < or = 0.0005). Taking together these latter two measures, an index of interaction was found to be 1.8 times higher when adult animals were compared to postweanling rats, and 2.5 times higher when compared to preweanling rats. Overall, these results are consistent with the view that there are late postnatal changes in the extent to which midbrain DA afferents interact specifically with GABAergic interneurons in rat mPFCx.

Differences in the subregional and cellular distribution of GABAA receptor binding in the hippocampal formation of schizophrenic brain.

Recent postmortem studies have reported a marked upregulation of GABAA receptor binding activity in the anterior cingulate and prefrontal cortices of schizophrenic subjects. Because the hippocampal formation is a key corticolimbic region that has also been implicated by both postmortem and brain imaging studies in the pathophysiology of this disorder, the current report has sought to determine whether alterations of GABAA receptor binding might also be detected in this region from 15 normal controls and 8 schizophrenic subjects. Using a low resolution autoradiographic approach, the results show a significant increase of specific GABAA receptor binding activity in the area dentata (granule cell layer), CA4, CA3 (str. oriens, str. pyramidale), subiculum, and presubiculum of the schizophrenic group. The magnitude of the increase was greatest in CA3 and lowest in the CA1 sector. When high resolution analyses were performed on emulsion-coverslip preparations, a modest increase of binding (43%, P = 0.05) was observed on pyramidal, but not non-pyramidal neurons in sector CA1. Rather unexpectedly, GABAA binding in sector CA3 was not significantly different on pyramidal cells, but was almost three-fold higher (P = 0.015) on non-pyramidal neurons of the schizophrenic group. There was no relationship of age or the postmortem interval to the parameters showing significant changes in the schizophrenic group. Moreover, patients both with and without neuroleptic exposure showed upregulation of GABAA receptor binding activity. Taking together the rather modest increase of binding activity in CA1 and the more marked upregulation in CA3, as well as the differential changes on pyramidal neurons of CA1 vs. non-pyramidal neurons in CA3, the findings reported here are consistent with the possibility that a disturbance of brain development could have occurred either perinatally or perhaps even well into the postnatal period, and have given rise to discreet subregional and cellular alterations of disinhibitory GABAergic modulation in sector CA3 of schizophrenics. Overall, the data reported here provide further evidence that alterations of GABAergic activity may occur in the hippocampal formation of schizophrenic patients.

Postnatal maturation of GABA-immunoreactive neurons of rat medial prefrontal cortex.

A light microscopic immunocytochemical approach has been used to examine the distribution and maturation of gamma-aminobutyric acid- (GABA) containing cells in rat medial prefrontal cortex (mPFC) at progressive postnatal stages. Between P1 and P5, labeled cells in the cortical plate show less differentiated morphological characteristics when compared to cells in the deeper laminae. By P10, however, most labeled cells in superficial laminae show more differentiated characteristics with some having a distinctive multipolar appearance. Between P1 and P5, there is a significant increase (50%) in the density of GABA-containing cells in the superficial laminae, while concurrently there is an overall decreases in the subjacent deeper laminae. As the cortex continues to expand, there is a corresponding decrease in the density of GABA-immunoreactive cells in the outer two-thirds of the cortical mantle until approximately P15, stabilizing at 20-25 cells/100,000 microns2 for all laminae. Between P1 and P15, there is also a significant increase (133%) in the average size of labeled cells, followed by a gradual decrease of 30% between P15 and P41. During P1-7, there is a marked increase in the density of labeled axosomatic terminals in both the superficial (200%) and deep laminae (116%). In the superficial layers, however, the density of labeled terminals again increases by 86% between P12 and P18. In general, the present findings are consistent with the idea that there is a progressive maturation of the intrinsic GABAergic system in rat mPFC in a classic "inside-out" pattern, and this involves extensive postnatal changes occurring during the first 3 postnatal weeks.

Cellular colocalization of dopamine D1 and D2 receptors in rat medial prefrontal cortex.

In a recent study in rat medial prefrontal cortex (mPFC), a fluorescently coupled, high-affinity ligand for the D1 receptor subtype was localized to nonpyramidal neurons, while a ligand selective for the D2 subtype was found on neurons with a size distribution overlapping with both small pyramidal and large nonpyramidal cells. These observations raised the possibility that a subpopulation of cortical neurons with an intermediate size range may coexpress both the D1 and D2 receptor subtypes. In the present study, the D1 and D2 receptor subtypes have been simultaneously localized in layer VI of rat mPFC using 20 nM SCH 23390-Bodipy and 20 nM N-(p-aminophenethyl) spiperone-Texas red, respectively, in the presence of 100 nM mianserin (5-HT2 receptor antagonist). The localization of receptor binding fluorescence was assessed in paired images using fluoroscein isothiocyanate (FITC) and rhodamine dichroic filters for the D1 and D2 subtypes, respectively. Under the conditions employed here, most cell bodies showed either D1-like or D2-like receptor binding fluorescence, while a colocalization of both fluoroprobes was observed on only 25% of the labeled cells. When the size of each single-labeled cell body was measured using the respective FITC (D1-probe) and rhodamine (D2-probe) epifluorescence filters, the distribution of cells showing only D1-like receptor binding fluorescence was similar to nonpyramidal neurons (68.6 +/- 1.8 microns 2), while that for cells showing only D2-like receptor binding fluorescence was similar to that of both large interneurons and small pyramidal cells (106.9 +/- 2.4 microns 2).(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of chronic haloperidol administration on GABA-immunoreactive axon terminals in rat medial prefrontal cortex.

Several reports have suggested that chronic haloperidol (HAL) treatment induces ultrastructural changes in synapses of substantia nigra, corpus striatum, and medial prefrontal cortex (mPFC) of rat brain. The effects of HAL on specific cortical transmitter systems, however, are not well characterized. Recent studies have indicated that there may be a loss of gamma-aminobutyric acid (GABA)ergic cells in anterior cingulate cortex of schizophrenic subjects and this hypothesis has prompted interest in the question of whether dopamine receptor antagonists, such as HAL, may influence the activity of this transmitter system. This current report describes a quantitative light microscopic analysis of GABA-immunolabeled axosomatic terminals in mPFC of rats treated with HAL decanoate (0.5 mg/kg/day, i.m.) for a period of 4 months. GABA-containing terminals were visualized with an avidin-biotin immunoperoxidase method for localizing anti-GABA antibodies. Computer-assisted image processing was employed to determine the total number of pixels representing GABA-immunoreaction product in axon terminals that were in direct apposition to pyramidal cell bodies. Drug-treated animals showed a significant increase in the number of pixels representing GABA-immunoreaction product in axosomatic terminals of layers II, III, VI, and VI (93%, 63%, 31%, and 43%, respectively). These data are consistent with the idea that chronic HAL administration may be associated with a significant increase in the amount of GABA present in terminals surrounding pyramidal neurons of rat mPFC. The fact that GABA-containing terminals showed the greatest increase in layer II is not consistent with the known distribution of dopamine afferents to this region which is lowest in superficial laminae. Based on the laminar distribution of non-dopaminergic receptor types that have a high affinity for HAL, the effect of this drug on GABAergic transmission could potentially involve changes that are mediated through mechanisms in which 5-HT2 or sigma opiate receptors play a role.

Dopamine-immunoreactive axon varicosities form nonrandom contacts with GABA-immunoreactive neurons of rat medial prefrontal cortex.

Recent postmortem studies have suggested that reduced gamma-aminobutyric acid (GABA)ergic activity in limbic cortex may be one component to the pathophysiology of schizophrenia. This hypothesis has underscored the importance of knowing whether midbrain dopamine afferents interact extensively enough with inhibitory interneurons to suggest a direct functional relationship. Toward this end, a double immunofluorescence approach combined with confocal laser scanning microscopy has been used to localize dopamine and GABA simultaneously in rat medial prefrontal cortex. The results confirm studies from other laboratories showing a rich network of dopamine-immunoreactive fibers forming a gradient across the cortical laminae, with deeper layers having the highest density. When viewed with oil immersion optics, dopamine-immunoreactive fibers were frequently found to be in close apposition with GABA-immunoreactive cell bodies. The percentage of GABA-containing neurons showing such contacts was highest in layer VI (65%) and progressively decreased toward layer I (9%). Varicose regions of the dopamine fibers were typically present at the point of contact with a GABA-immunoreactive cell body. Using an immunoperoxidase technique to localize dopamine fibers and cresyl violet staining to visualize neurons simultaneously, two separate statistical analyses were performed to assess whether the frequency of contacts between dopamine fibers and cell bodies in general may be due to random effects. In layer VI, a high percentage of both pyramidal and nonpyramidal neurons were found to be in contact with dopamine varicosities (71% and 76%, respectively), but these were not significantly different from that observed for GABA-containing cells (65%) in double-immunofluorescence specimens. A Chi-square statistical test was used to compare the observed and predicted number of varicosities forming cell body contacts. This analysis indicated that the percentage of dopamine varicosities (30%) that form appositions with cell bodies is much greater than would be expected if these appositions were due to random effects (15%). Moreover, using an estimate of intensity for a stationary Poisson process, it was again found that random effects can not account for these interactions (P = 0.01). Taken together with earlier electron microscopic studies from other laboratories, the present findings support the idea that GABAergic interneurons have extensive interactions with dopamine varicosities. While these interactions are not unique to GABAergic cell bodies, they suggest that inhibitory interneurons can play a direct role in mediating the effects of midbrain dopamine afferents in rat medial prefrontal cortex.

Cellular distribution of dopamine D1 and D2 receptors in rat medial prefrontal cortex.

The relative distribution and cellular localization of the dopamine D1 and D2 receptor subtypes were assessed in frozen sections of rat medial prefrontal cortex (mPFC). The D1 and D2 receptor binding sites were labeled with the selective high-affinity antagonists SCH 23390 and N-(p-aminophenethyl)-spiperone (NAPS), respectively, coupled to either Bodipy or Texas red fluorophores. Under the incubation conditions employed, kinetic, competition, and selectivity studies showed that these modified ligands retained pharmacological selectivity. Optimal binding fluorescence was at 100 nM of each ligand, and fluorescence increased linearly from 1 to 15 min of incubation at 2 degrees C. NAPS-Texas red binding fluorescence was inhibited with 10 nM quinpirole (D2 agonist), but not 10 nM SKF 38393 (D1 agonist), while SCH 23390-Texas red binding was inhibited with SKF 38393, but not quinpirole. The localization of dopamine receptor binding was assessed in montages constructed from low-magnification photomicrographs through the depth of the cortex, or in corresponding high-magnification photomicrographs. Cells showing D1- or D2-like receptor binding fluorescence were present in layers II-VI, with the highest density observed in layers V and VI. The addition of mianserin (100 nM, 5-HT2 antagonist) to incubated sections slightly reduced the numbers of labeled cells in each cortical layer, but retained the preferential localization to the deeper layers. Two separate observations supported the idea that the fluorescently coupled ligands were localized to neuronal cell bodies. First, receptor labeling with the fluorescently coupled ligands co-localized almost exclusively to cells in the cortical mantle showing neuron-specific enolase immunoreactivity. Second, a comparison of the cell size distribution taken from adjacent Nissl-stained sections with the size of cells showing D1- or D2-like receptor binding fluorescence revealed complete overlapping of fluorescence with neuronal cell bodies. In mPFC layer VI, the size of cells showing D1-like receptor binding fluorescence was 77.8 +/- 5.1 microns2, similar to non-pyramidal neurons, while that for D2-like receptor binding fluorescence was 108.2 +/- 4.5 microns2, consistent with both large interneurons and small pyramidal cells. Only a small percentage of cells showing D1- or D2-like receptor binding overlapped in size with glia, but this occurred almost exclusively within the white matter region below the cortical mantle. These findings are consistent with the hypothesis that the D1 and D2 receptor subtypes are found on different populations of neurons, although some overlap probably occurs.(ABSTRACT TRUNCATED AT 400 WORDS)

Increased density of glutamate-immunoreactive vertical processes in superficial laminae in cingulate cortex of schizophrenic brain.

Recent postmortem investigations have suggested that schizophrenia may involve a defect in associative information processing in the upper layers of limbic cortex. One of these studies reported that vertical processes visualized with antibodies against the neurofilament 200K subunit (NFP-200K) of the axon cytoskeleton were increased in density in layer II and upper portions of layer IIIa of the cingulate region of schizophrenic individuals. Based on this latter finding, it was hypothesized that there may be a superbundance of associative afferents to this region. To explore this possibility further, an immunoperoxidase localization of the amino acid glutamate has been employed to visualize vertical fibers in layers II and IIIa of postmortem anterior cingulate cortex in both normal controls (n = 15) and schizophrenics (n = 17). Vertical fibers were distinguished according to small or large calibers and were differentially counted with a blind computer-assisted technique. The schizophrenic group showed a markedly higher density (77.8%) of small-caliber glutamate-immunoreactive vertical fibers when compared to controls; the density of large-caliber vertical fibers also showed a similar, though smaller (30.2%), increase in the schizophrenic group. There were no differences in the density of either small- or large-caliber processes in prefrontal cortex of the two groups. The effects of age, postmortem interval, fixation, and neuroleptic exposure do not account for the differences between the normal and schizophrenic subjects. Taking together their small caliber, vertical orientation, localization in superficial layers, and marked glutamate immunoreactivity, it seems plausible that the fibers showing an increased density in schizophrenics may be glutamatergic afferents, possibly ones that are associative in nature.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased GABAA receptor binding in superficial layers of cingulate cortex in schizophrenics.

Recent investigations of postmortem brain from schizophrenic patients have revealed reduced numbers of neurons in several different corticolimbic brain regions. In the prefrontal and anterior cingulate cortices, more specific decreases in the numbers of interneurons, but not pyramidal cells, have been reported to occur preferentially in layer II. Based on this latter finding, a loss of inhibitory basket cells leading to a compensatory upregulation of the GABAA receptor has been hypothesized to occur in schizophrenic patients and to be a contributory factor in the pathophysiology of this disorder. We now report the results of a high-resolution quantitation of GABAA receptor binding in anterior cingulate cortex of postmortem specimens from normal and schizophrenic cases. The results indicate a preferential increase in bicuculline-sensitive 3H-muscimol binding on neuronal cell bodies of layers II and III, but not layers V and VI, of the schizophrenic cases. There was no difference in the size of neurons in any of the layers examined when the control and schizophrenic groups were compared. The neuropil of layer I also showed significantly greater GABAA binding in schizophrenics. The differences seen in the schizophrenic group did not appear to be the result of exposure to antipsychotic medication because one patient who was medication naive and a second who had received minimal exposure to antipsychotic drugs also showed elevated GABAA receptor binding. Since information processing depends on corticocortical integration in outer layers I-III, a disturbance of inhibitory activity in these superficial layers of limbic cortex may contribute to the defective associative function seen in schizophrenia.

Localization and high-resolution imaging of cortical neurotransmitter compartments using confocal laser scanning microscopy: GABA and glutamate interactions in rat cortex.

This report compares the application of confocal laser scanning fluorescence microscopy with standard epifluorescence microscopy for the simultaneous localization of the neurotransmitters gamma-aminobutyric acid and glutamate in rat cerebral cortex. With this approach, sections of fixed rat brain are treated with primary antibodies against gamma-aminobutyric acid (rabbit-derived) and glutamate (mouse-derived), followed by treatment with fluorescein isothiocyanate-tagged donkey anti-rabbit and rhodamine-tagged goat anti-mouse secondary antibodies, respectively. The results demonstrate that images from immunofluorescence localizations with a confocal laser scanning microscope have superior resolution and contrast as a result of significant reductions of background flare caused by emission from out-of-focus structures in the field of view. The confocal microscope achieves this improved image quality by optically sectioning through a specimen at narrow planes of focus and then compiling a composite image of an object of interest. The composite image can be further enhanced by using various image processing options. The combined use of double immunofluorescence and confocal laser scanning microscopy provides an important means to simultaneously study the anatomical relationships of pre- and post-synaptic elements in a complex neural system.

Deficits in small interneurons in prefrontal and cingulate cortices of schizophrenic and schizoaffective patients.

A recent report suggested that neurons in the prefrontal, anterior cingulate, and primary motor cortex of the brains of schizophrenic subjects may be less dense than those in the brains of nonschizophrenic subjects. We have determined whether pyramidal neurons and/or interneurons are preferentially reduced in schizophrenic subjects. Twelve control subjects and 18 schizophrenic subjects were studied in a blind, quantitative analysis of the density of pyramidal cells, interneurons, and glial cells in each of the six layers of the anterior cingulate and prefrontal cortex. The results showed that numbers of small neurons (interneurons) were reduced in most layers of the cingulate cortex in schizophrenic subjects compared with nonschizophrenic subjects, with the differences being greatest in layer II. In the prefrontal area, interneuronal density was also lower in layer II and, to a lesser extent, in layer I in schizophrenic subjects compared with control subjects. In most cases, the differences were similar, although more significant, in schizophrenic subjects who had had superimposed mood disturbances than in schizophrenic subjects who had not had such comorbidity. Numbers of pyramidal neurons generally were not different between control and schizophrenic subjects, except in layer V of the prefrontal area, where schizophrenic subjects showed higher densities of these neurons. Glial numbers did not differ between the control and schizophrenic subjects, suggesting that a neurodegenerative process did not cause the reduced interneuronal density observed. Using multiple regression analysis and analysis of covariance, decreases in the density of layer II interneurons could not be adequately explained by the effects of various confounding variables, such as age, postmortem interval, duration of specimen fixation, or administration of neuroleptic agents.(ABSTRACT TRUNCATED AT 250 WORDS)

Evidence for ultrastructural changes in cortical axodendritic synapses following long-term treatment with haloperidol or clozapine.

A quantitative electron microscope analysis was performed to determine the effects of 1-year administration of either haloperidol or clozapine on the ultrastructure of synapses in layer VI of rat medial prefrontal cortex. The average caliber of dendritic shafts was significantly decreased in the haloperidol-treated group, but not in clozapine-treated animals; however, there was no difference between the means of the two drug-treated groups. Both drug-treated groups showed a significant decrease in the relative distribution of axodendritic synapses displaying asymmetric postsynaptic membrane specializations, while there was an increase in synapses with either a symmetric membrane specialization or no specialization at all. Axospinous synapses of the clozapine-treated group showed a shift in the relative distribution of synaptic subtypes paralleling that found for axodendritic synapses. There was no change in the number of axon terminals forming a synapse on dendritic cross-sectional profiles in the neuroleptic-treated groups. Similarly, no change in the average size of axon terminals or synaptic vesicle density was detected. These data suggest that both typical and atypical neuroleptics, when administered over an extended period of time, may induce relative shifts in the distribution of excitatory and inhibitory elements synapsing on dendritic shafts and, to a lesser degree, spines. Such alterations in synaptic relationships may provide insight into the interaction of neuroleptics with intrinsic components in layer VI of medial prefrontal cortex.

Effects of aging on the neuroglial cells and pericytes within area 17 of the rhesus monkey cerebral cortex.

An electron microscopic analysis has been carried out to compare the neuroglial cells and pericytes within the primary visual cortex, area 17, of young (5-6 years) and old (25-35 years) rhesus monkeys. All of the neuroglial cell types accumulate inclusions within their cytoplasm as they age, and the inclusions within the astrocytes and oligodendrocytes are essentially characteristic of those cell types. The astrocytes probably acquire their inclusions by phagocytosis, and it is suggested that the inclusions in the oligodendrocytes are caused by an age-related degeneration of the myelin sheaths they produce. The inclusions within the microglia are very heterogeneous. They are more massive than in the other neuroglial cells, so that their inclusions may almost fill the microglia. Pericytes also accumulate inclusions with age and there is evidence to suggest that they empty the contents of their inclusions vacuoles directly into the capillaries. On the basis of counts of the numbers of profiles of neuroglial cells displaying nuclei in thin sections, the only cells to increase in number with age are the microglia. They show an increase of about 44% when the cortices of young and old monkeys are compared.

The rat parvocellular reticular formation: I. Afferents from the cerebellar nuclei.

The present report elucidates the location of the cells of origin of the cerebellar nuclear-parvocellular reticular projection in the rat. Injection of horseradish peroxidase (HRP) into the parvocellular reticular formation resulted in labeled neurons in the dorsolateral protuberance of the contralateral medial nucleus and bilaterally in the dorsolateral hump region and the large celled subgroup of the lateral nucleus. A small number of labelled cells were found in the middle part of the contralateral medial nucleus. The few HRP-positive cells in the interpositus nuclei were adjacent to the dorsolateral hump. No labeled neurons were observed in the caudomedial portion of the medial nucleus or the small-celled region of the lateral nucleus.

High resolution imaging of receptor binding in analyzing neuropsychiatric diseases.

A method for analyzing high resolution imaging of receptor binding activity in human post-mortem brain specimens is described. The autoradiography technique employed is based on methods previously described by others in which coverslips dipped in tritium-sensitive nuclear track emulsion are placed over a tissue section that has been incubated in a medium containing radioactively-tagged ligand. With this approach, there is a 370-fold increase in resolution from approximately 120 microns available with tritium-sensitive films to 0.33 micron attainable with the emulsion approach. Since the coverslip autoradiogram remains superimposed on the tissue section, individual grains can be routinely quantitated in specific cell types and discrete subregions of the neuropil with the aid of a user-interactive image processing system. Overall, the improved resolution that this approach provides makes it possible to determine whether a particular neuronal sub-type may be preferentially altered by disease processes affecting the brain.