Cortical Neuronal Density Research Articles

Injectable Dexamethasone Administration Enhances Cortical GABAergic Neuronal Differentiation in a Novel Model of Postnatal Steroid Therapy in Mice

Injectable dexamethasone (DXM) is widely used during the postnatal period in premature infants. However, this treatment has been associated with an increased incidence of neuromotor disorders. Few studies have directly addressed the impact of DXM therapy on neuronal differentiation. We used a murine model of postnatal steroid therapy in which mouse pups aged 3 and 4 postnatal days (P) received intraperitoneal injections of 1 mg . kg(-1) . 12 h(-1) of an injectable preparation that contained DXM and sulfites (DXM), pure DXM, or sulfites. The animals were weighed before they were killed on P5, P10, or P21, and their brains were investigated by immunohistochemistry with markers for neuronal differentiation. DXM administration was associated with a 20-30% reduction in body and brain weight gains and in cortical thickness on P5 and P10. gamma-Amino-butyric acid+ (GABA+) interneuron density was significantly increased (+50%) in the cerebral cortex of the animals given injectable DXM on P5 to P21 compared with controls (p < 0.01). In parallel, the density of cortical neurons expressing two interneuron markers (calbindin 28-kD and calretinin) increased significantly. These alterations occurred with injectable DXM but not with pure DXM or sulfites alone. In contrast, none of the study treatments modified the expression of other markers for neuronal transmission or axon myelination. In the animals that were given injectable DXM, cleaved caspase 3 antibody showed increased neuronal cell death, but calbindin antibody did not. In conclusion, in a murine model of postnatal steroid therapy, injectable DXM induced a selective increase in GABAergic neurons in the cerebral cortex.

The Effects of Decompression and Exogenous NGF on Compressed Cerebral Cortex

Using a rat epidural bead implantation model, we found that compression alone could reduce the overall and individual layer thicknesses of cerebral cortex with no apparent cell death. The dendritic lengths and spine densities of layer II/III and V pyramidal neurons started to decrease within 3 days of compression. Decompression for 14 days resulted in near complete to partial recovery of the cortical thickness and of the dendritic lengths of layer II/III and V pyramidal neurons, depending on the duration of the preceding compression. The recoverability was better following short (3-day) than long (1- or 3-month) periods of compression. The loss of dendritic spines nevertheless persisted. An intraventricular infusion of NGF was performed after decompressing the lesions following 3 days of cortical compression, and this increased the recovery of the spines but not the dendritic length of the cortical pyramidal neurons, nor did it alter the recovery of the cortical thickness. NGF also promoted the increase of the dendritic spines, but not the dendritic length of the cortical pyramidal neurons of normal animals. In short, the data show that a few days of compression alone can cause permanent cortical damage. Exogenous NGF, if applied topically, may restore the dendritic spine density of cortical neurons subjected to compression.

A quantitative morphometric study of the human anterior cingulate cortex

Morphometric alterations in the anterior cingulate cortex (ACC) have been reported in schizophrenia and mood disorders. Parameters affected include glial and neuronal density, neuronal size and cortical thickness. Some data, especially in mood disorders, suggest that the left subgenual ACC is preferentially involved. Qualitative studies show that the ACC cytoarchitecture is heterogeneous, but there are few quantitative data. We performed a quantitative morphometric study of five anatomical levels within the ACC (caudal and rostral subgenual [area 24b sg], rostral and intermediate supragenual [area 24b] and caudal supragenual [area 24b′]) in both hemispheres of five normal brains. We measured cortical depth, layer depths, neuronal density, neuronal size, and glial density, using the optical disector and nucleator. Relative to the subgenual ACC, the supragenual ACC was thicker, with a deeper layer V. Supragenual neurons were substantially larger in all layers, and were less densely packed in layers V and VI, than subgenual neurons. Glial density, and the glia to neuron ratio, was higher in supragenual than subgenual ACC. Only minor differences were seen between left and right ACC, between caudal and rostral subgenual ACC, and between the three supragenual levels. These data complement the qualitative descriptions of the heterogeneity of human ACC cytoarchitecture, connections, and functions, especially between supragenual and subgenual regions. Our findings also indicate that care must be taken when selecting ACC tissue to be used for morphometric studies of psychiatric disorders, since the normal anatomical variation is of a similar magnitude as the reported disease-related alterations.

A quantitative morphometric study of the human anterior cingulate cortex

Morphometric alterations in the anterior cingulate cortex (ACC) have been reported in schizophrenia and mood disorders. Parameters affected include glial and neuronal density, neuronal size and cortical thickness. Some data, especially in mood disorders, suggest that the left subgenual ACC is preferentially involved. Qualitative studies show that the ACC cytoarchitecture is heterogeneous, but there are few quantitative data. We performed a quantitative morphometric study of five anatomical levels within the ACC (caudal and rostral subgenual [area 24b sg], rostral and intermediate supragenual [area 24b] and caudal supragenual [area 24b′]) in both hemispheres of five normal brains. We measured cortical depth, layer depths, neuronal density, neuronal size, and glial density, using the optical disector and nucleator. Relative to the subgenual ACC, the supragenual ACC was thicker, with a deeper layer V. Supragenual neurons were substantially larger in all layers, and were less densely packed in layers V and VI, than subgenual neurons. Glial density, and the glia to neuron ratio, was higher in supragenual than subgenual ACC. Only minor differences were seen between left and right ACC, between caudal and rostral subgenual ACC, and between the three supragenual levels. These data complement the qualitative descriptions of the heterogeneity of human ACC cytoarchitecture, connections, and functions, especially between supragenual and subgenual regions. Our findings also indicate that care must be taken when selecting ACC tissue to be used for morphometric studies of psychiatric disorders, since the normal anatomical variation is of a similar magnitude as the reported disease-related alterations.

Differential effects of cholinergic lesions on dendritic spines in frontal cortex of young adult and aging rats

Previously, we demonstrated that plasticity of frontal cortex is altered in aging rats: cholinergic lesions of the nucleus basalis magnocellularis (NBM) produce larger declines in dendritic morphology in frontal cortex of middle-aged and aged rats relative to young adults. To more closely examine the interactive effects of age and cholinergic deafferentation on synaptic connectivity in frontal cortex, we assessed the effects of specific cholinergic lesions on spine density of frontal cortical neurons in young adult, middle-aged, and aged rats. Rats received unilateral sham or 192 IgG-saporin lesions of the NBM. Two weeks after surgery, brains were stained using a Golgi–Cox procedure, and spine density was quantified in second-, third-, and fourth-order basilar dendrites of pyramidal neurons in layer II–III of frontal cortex. Spine density was reduced at all branch orders in aged, sham-lesioned rats. In addition, whereas lesions produced a marked increase in spine density on second- and third-order branches in young adult rats, lesions failed to significantly alter spine density in middle-aged and aged rats. Thus, the upregulation of dendritic spines may be a compensatory response to deafferentation, which is lost with advancing age.

Kainate-induced epileptic seizures induce a recruitment of caldendrin to the postsynaptic density in rat brain

Caldendrin defines a novel family of neuronal calcium-sensor proteins, the C-terminal moiety of which displays high similarity to calmodulin. We now report that the protein is recruited to the postsynaptic density (PSD) of cortical and hippocampal neurons in response to kainate-induced epileptic seizures, an animal model of human temporal lobe epilepsy. The translocation of caldendrin to the PSD did not occur in kainate-treated rats that did not develop seizures. The enhanced PSD levels of caldendrin are not due to increased protein synthesis and most likely reflect a recruitment from the soluble caldendrin protein pool. These findings suggest that the transduction of dendritic Ca 2+-signals via caldendrin is altered by epileptic seizures and that caldendrin might be involved in the pathophysiology of temporal lobe epilepsy.

Cell pathology in mood disorders.

In new, exciting, neuroanatomic studies on postmortem tissues from patients with mood disorders, quantitative cytomorphologic differences can be shown at the microscopic level. These investigations provide direct evidence that mood disorders are characterized by marked reductions in glial cell number and density in addition to subtle alterations in the density and size of cortical neurons in frontolimbic brain regions. Importantly, this corresponds with clinical neuroimaging studies and preclinical animal studies that suggest cell atrophy, cell loss, or impairments in neuroplasticity and cellular resilience may underlie the neurobiology of major depressive disorder and bipolar manic-depressive disorder. Because this represents a departure from modern efforts to understand mood disorders, published reports are scarce and based on rather small sample sizes. This article reviews the current findings from postmortem studies on glial and neuronal cell counts in primary mood disorders and discusses a possible link between cellular changes and the action of psychotherapeutic drugs.

Primates exposed to cocaine in utero display reduced density and number of cerebral cortical neurons.

This study examined the effects of cocaine use during the second trimester of pregnancy on cerebral neocortical volume and density, and total number of neocortical neurons and glia in offspring. We also evaluated the extent of postnatal recovery of cytoarchitectural abnormalities previously observed in the neocortex of two-month-old primates born from cocaine-treated mothers (Lidow [1995] Synapse 21:332-334). Pregnant monkeys received cocaine orally (20 mg/kg/day) from the 40th to 102nd days of pregnancy (embryonic day [E]40-E102). On E64 and E65, the animals were injected with [(3)H]thymidine. Cerebral hemispheres of the offspring were examined at three years of age. We found a reduction in the neocortical volume and density and total number of neocortical neurons. The observed reduction in neuronal number within the neocortex was not accounted for by the increase in the number of neurons in the white matter of cocaine-exposed animals, because the number of these "extra" neurons was equal to only half that of missing neurons. We detected no significant changes in the number of neocortical glia. The cytoarchitectural abnormalities in the neocortex of prenatally cocaine-exposed three-year-old monkeys closely resembled previously described neocortical abnormalities in similarly exposed two-month-old animals: the neocortex lacked a discernible lamination; the majority of the cells labeled by [(3)H]thymidine injected during neocortical neurogenesis did not reach their proper position within the cortical plate. Therefore, postnatal maturation is not associated with significant improvement in neocortical organization in primates prenatally exposed to cocaine. There was, however, a postnatal recovery of low glial fibrillary acidic protein (GFAP) immunoreactivity previously observed in 2-month-old cocaine-exposed animals.

Cocaine affects cerebral neocortical cytoarchitecture in primates only if administered during neocortical neuronogenesis

Previously, we demonstrated that chronic exposure of fetal monkeys to cocaine could result in development of the neocortex with significant cytoarchitectonic abnormalities [Synapse, 21 (1995) 435–444]. In the present study, we examined the developmental time-frame within which neocortical cytoarchitecture is susceptible to modifications by prenatal cocaine exposure. For this purpose, we assessed the integrity of cortical lamination and the position, density, and total number of occipital cortical neurons in 2-month-old monkeys which were prenatally exposed to chronic cocaine treatment either prior to the period of neocortical neuronogenesis, during the period of neocortical neuronogenesis, or after the period of neocortical neuronogenesis. We found that cocaine can interfere with the neocortical laminar organization and induce a reduction in the density and number of neocortical neurons only if it is administered at the time of neocortical neuronogenesis. During this window of vulnerability, an abnormal neocortex is generated as long as cocaine exposure is maintained, with corticogenesis becoming normal as soon as the administration of this drug is discontinued.

The amyloid paradox resolved — thioflavine positive abeta deposits lead to local neuropil damage and neuronal death

Cortical microglia exhibit a ramified shape during sleep, while they have a hyper-ramified shape during wakefulness, which is characterized by their longer processes with increased branching points. The microglial molecular circadian clock regulates expressions of both cathepsin S (CatS) and P2Y12 receptors in the brain with a peak at zeitgeber time 14 (2 h after beginning of the dark phase). We postulated that these two microglia-specific molecules contribute to diurnal alterations of microglial shapes and neuronal activities in the cerebral cortex. During wakefulness, CatS secreted from cortical microglia may be involved in P2Y12 receptor-dependent process extension. Secreted CatS subsequently degrades the perineuronal nets, initiating the downscaling of both spine density and synaptic strength of cortical neurons toward the beginning of sleep. The downscaling of both spine density and synaptic strength of cortical neurons during sleep could improve signal-to-noise, which would benefit memory consolidation, or allow for new learning to occur during subsequent waking. Furthermore, disruption of CatS induces the sleep disturbance and impaired social interaction in mice. Moreover, the microglial clock system disruption may also play a role in the early pathogenesis of Alzheimer's disease. The reduced expression of BMAL1 in cortical microglia caused by oligomeric amyloid β may induce the increased presence of inflammatory phenotype through a reduction in RORα, which in turn reduced IκBα and enhanced NF-κB activation.These observations suggest that the microglial clock system disruption contribute to pathogeneses of sleep disturbance, impaired social interaction and cognitive impairment. Therefore, the growing understanding of the microglial circadian molecular clock might aid in the development of novel pharmacological interventions against both neuropsychiatric and neurodegenerative disorders.

Microdysgenesis with abnormal cortical myelinated fibres in temporal lobe epilepsy: a histopathological study with calbindin D-28-K immunohistochemistry.

Microdysgenesis is a microscopic cortical malformation reported to occur with varying incidence in surgical lobectomies from patients with temporal lobe epilepsy (TLE). It may act as a substrate for the seizures. Four patients are reported with TLE, hippocampal sclerosis and cortical microdysgenesis which was also characterized by the presence of abnormal myelinated fibres running tangentially in the superficial cortical laminae and closely associated with abnormal clusters of neurones. Similar abnormal cortical fibres have been described in other malformations of cortical development including polymicrogyria and focal cortical dysplasia and it is therefore likely that these fibres represent part of the microdysgenetic malformation not hitherto reported. The possibility is discussed that they may also be of functional significance in terms of influencing local seizure propagation and the secondary cortical neuronal loss observed, predominantly affecting layer II. Studies of calbindin interneuronal populations showed preservation of these cells in the microdysgenetic cortex, when compared with non-malformed temporal lobes, despite an overall reduction in cortical neuronal density. In addition, prominent numbers of neurogliaform calbindin-positive nerve cells were observed in the microdysgenesis cases and the nature of these cells is speculated upon.

Increased TUNEL staining in brains of autoimmune Fas-deficient mice

Profound changes in brain morphology and behavior coincide with the spontaneous development of systemic autoimmune/inflammatory disease in Fas-deficient MRL-lpr mice. The dendrites atrophy, the density of hippocampal and cortical neurons decreases, and an anxious/depressive-like behavior emerges while lymphoid cells infiltrate into the choroid plexus of MRL-lpr mice. We hypothesized that the inherited lack of the Fas-dependent anti-inflammatory mechanism would lead to unsuppressed immune activity, characterized by reduced apoptosis in the MRL-lpr brain. Using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeled (TUNEL) method as an indicator of apoptosis, a surprisingly high incidence of TUNEL-positive cells was observed in the hippocampus, choroid plexus and periventricular regions of MRL-lpr mice, 5–10-fold higher than that found in the MRL +/+ control brain. Immunostaining with anti-CD3, CD4 and CD8 monoclonal antibodies showed limited overlap between CD-positive and TUNEL-positive cells, suggesting that the dying cells are for the most part (∼70%) not T-lymphocytes. Although further characterization of the phenotype of the dying cells and the mechanism of cell death are required, the present results suggest the involvement of a Fas-independent apoptotic process in neurodegeneration induced by systemic autoimmune disease.

Sparse-sampling of gratings in the visual cortex of strabismic amblyopes

Strabismic amblyopes show losses in positional acuity that cannot be explained by their resolution or contrast sensitivities. One hypothesis for these losses is a reduction in the density of cortical neurons that are driven by the amblyopic eye (cortical undersampling). The question this study addressed was whether the foveal representation of the amblyopic eye is undersampled in the cortex of strabismic amblyopes. In order to assess spatial sampling psychophysically, we recorded the perceived orientation of a stationary grating as a function of grating orientation and frequency in three strabismic amblyopes. To ensure high retinal contrast, the grating was imaged on the fovea of each observer using a laser interferometer. We found that the strabismic amblyopes misperceived the orientation of the grating at spatial frequencies that are a factor of two to six lower than the sampling frequency of the foveal cones. Since the retina and LGN in strabismic amblyopes are presumably normal, this result suggests sparse cortical sampling in the foveal representation of the amblyopic eye. Undersampling by cortical neurons may contribute to the spatial distortions present in strabismic amblyopic eyes.

Effects of mild protein prenatal malnutrition and subsequent postnatal nutritional rehabilitation on noradrenaline release and neuronal density in the rat occipital cortex

There is evidence that severe malnutrition started during gestation and continued through lactation affects adversely the morphologic development of the neocortex leading to increased neuronal cell packing density and decreased dendritic branching. Nevertheless, the effect of purely mild protein prenatal malnutrition on neocortical development remains rather unexplored. This study evaluates the effects of mild protein prenatal malnutrition (8% casein diet, calorically compensated by carbohydrates) and subsequent postnatal nutritional rehabilitation (25% casein diet) on: (i) the potassium-induced release of [ 3 H ]-noradrenaline (NA) in occipital cortex slices obtained from 1- and 22-day-old pups; and (ii) the packing density of neurons in lateral, dorso-lateral and dorsal regions of the occipital cortex of 22-day-old pups by using the optical dissector method. The experiments were performed in rats normally fed during gestation and lactation (G +L +), malnourished during gestation but rehabilitated during lactation (G −L +) and malnourished during gestation and lactation (G −L −). At day 1 of age, no significant differences in body and brain weights were observed between prenatally well-nourished and malnourished pups. Nevertheless, at this early age, pups born from mothers submitted to the 8% casein diet had significantly higher cortical net percent NA release than pups born from mothers receiving the 25% casein diet. At weaning (22 days of age) G −L + rats had, compared to the G +L + group, similar body weight, brain weight and [ 3 H ]-NA release values, but significantly higher neuron density scores in the lateral region of the occipital cortex. In contrast, at 22 days of age, G −L − rats exhibited, compared to G +L + animals, significant deficits in body and brain weights as well as significant increases in cortical net percent NA release together with enhanced packing density of neurons in the lateral, dorso-lateral and dorsal regions of the occipital cortex. Moreover, in G −L − animals was not found the laterodorsal histogenetic gradient of neuronal cell packing density observed in G +L +rats. Results suggest that mild prenatal malnutrition per se is able to induce deleterious effects on cortical neuronal density, in spite of nutritional rehabilitation during lactation, through a mechanism involving central NA hyperactivity during gestation. Prosecution of malnutrition after birth magnifies both neurochemical and morphometric disorders.

Area- and lamina-specific organization of a neuronal subpopulation defined by expression of latexin in the rat cerebral cortex

The aim of the present study was to investigate the density, laminar distribution, size, morphology, and neurotransmitter phenotype of rat cortical neurons expressing latexin, an inhibitor of carboxypeptidase A. Immunohistochemical analyses established that latexin-immunoreactive neurons are restricted essentially to the infragranular layers of lateral cortical areas in the rat. The overall density, laminar or sublaminar localization, and cell size distribution of latexin-positive neurons differed substantially across cytoarchitectonic areas within lateral cortex. Numerous latexin-positive neurons had the morphology of modified pyramidal cells especially of layer VI. The vast majority of latexin-positive neurons were glutamate-immunoreactive in the six lateral neocortical areas examined, while neurons immunoreactive for both latexin and GABA were virtually absent. Thus the majority of latexin-positive neurons are likely to be excitatory projection neurons. The area- and lamina-specific distribution of the latexin-expressing subpopulation of glutamate-immunoreactive neurons is a distinctive feature that may contribute to the functional specialization of the lateral cortical areas.

Characterization of neuronal damage by iomazenil binding and cerebral blood flow in an ischemic rat model.

I-123-iomazenil is a SPECT probe for central benzodiazepine receptors (BZR) which may reflect intact cortical neuron density after ischemic insults. We evaluated whether neuronal damage in rats could be characterized by iomazenil as compared with cerebral blood flow (CBF). Serial changes in I-125-iomazenil for BZR and I-123-IMP for CBF were analyzed after the unilateral middle cerebral artery occlusion in rats by using an in vivo dualtracer technique. Uptake ratios of affected to contralateral regions were calculated. The iomazenil as well as IMP were decreased in all regions except for the cerebellum (remote area). Both iomazenil and IMP increased over time except in the temporal region (ischemic core). The iomazenil uptake was higher than IMP except in the ischemic core between 1 and 3-4 wk when iomazenil was lower than IMP. Iomazenil showed a moderate decrease in the proximal and middle parietal regions (peri-infarct areas) at 3-4 wk. The triphenyl-tetrazolium-chloride (TTC) stain at 1 wk demonstrated unstained tissue in the temporal region indicating tissue necrosis. With hematoxylin-eosin (HE) stain at 1 wk, widespread neuronal necrosis with occasional intact neurons were found in the proximal parietal region, and isolated necrotic neurons were represented in the distal parietal region. Iomazenil correlated well with the neuron distribution and the finding of a discrepancy between iomazenil and IMP might be useful in evaluating the neuronal damage.

Evidence for a Postnatal Doubling of Neuron Number in the Developing Human Cerebral Cortex Between 15 Months and 6 Years

The generalization of the finding of no postnatal neurogenesis in non-human primates to humans may be incorrect because: (1) rhesus macaques belong to a superfamily that diverged more than 25 million years ago from the superfamily including the genus Homo; (2) the pulse thymidine labeling method, which demonstrates DNA synthesis rather than mitosis per se, is less reliable than some have assumed. This study examines changes in the number of neurons in a column underneath a cortical surface area of 1 mm2, extending through all cortical layers (mm2-column) for 35 gyri (representing about 73% of the human cerebral cortex) based on the data of J.L. Conel (1939 to 1967). We corrected these data, derived from his measures of cortical neuronal packing density, somal breadth and height, and cortical layer thickness at postnatal ages 0, 1, 3, 6, 15, 24, 48, and 72 months, for shrinkage and stereological errors. In all 35 gyri, neuron number/mm2-column: (1) initially declines (μ=46% decline, σ=8%), 95% of which is due to surface area expansion (mean age of nadir value=15.8 months); (2) then increases to age 72 months by 70% (μ=1.7-fold increase, (μ rate=1.1% per month). Because of a concomitant 1.3-fold increase in cortical surface from 15 to 72 months, total cortical neuron number increases 2.2-fold. The close agreement between neuron number/mm2-column for Comel's age 72-month data to the corresponding values reported by others for adult human and primate cortex using more modern methods suggests the finding is not an artifact. Neuronal proliferative fate-determining factors provide at least four mechanisms for increasing cortical neuron number postnatally, with or without DNA synthesis.

Morphometric peculiarities of the brain in two-month-old rats as a function of body weight at the age of one month

The brain from 2-month-old rats up to the age of 1 month was studied in litters of different size: 9–13 (control) and 4–6 (reduced) animals. In animals from reduced litters the absolute weight of the brain surpassed that of controls. The maximum brain weight is characteristic of animals from reduced litters with maximum body weight at the age of one month, while the minimal brain mass is found in controls with the minimal body weight. There are no differences in the cortical thickness and neuron density in layers II and V between these two groups.

Laminar distribution of neuropeptide Y-immunoreactive neurons in human prefrontal cortex during development.

Neuropeptide Y (NPY) is present in neurons of the adult human cerebral cortex. In view of the reported roles of NPY in the central nervous system in health and during certain disease conditions, we have studied normal development of NPY immunoreactivity (-ir) in the human prefrontal cortex (PFC), Brodmann areas 9 and 46. Twenty-six specimens ranging from the ages of 14 postovulatory weeks to 34 years exhibited patterns that revealed six periods in the development of the laminar distribution and density of NPY-ir neurons. Changes during prenatal and perinatal periods reflect the onset, development, and resolution of the transient fetal telencephalic compartments, including the subplate zone, in which NPY-ir neurons are especially abundant. Before the age of 1 year, the majority of NPY-ir neurons were found in the subplate zone, whereas, after 1 year, the majority were seen in the cortical layers. This is in contrast with the human visual cortex, where the majority of NPY-ir neurons were still located in the white matter. The density of cortical NPY-ir neurons increased in the fifth developmental period (ages 4-7 years), coinciding with the increase of cortical volume and marked progression of cognitive functions. The adult pattern of a relatively low density of cortical NPY-ir neurons was reached in period 6 (from about 8 years), when individual variation also became apparent. Our data point to a protracted maturation of NPY-ir in the human PFC and to different distribution patterns of NPY-ir neurons in different cortical areas.

Light microscopic quantification of morphological changes during aging in neurons and glia of the rat parietal cortex.

Different changes in neuronal and glial population of the aging brain have been described; however, the degree and extent of these changes are controversial. This study evaluates the quantitative and cytomorphometric effects of aging on neuronal and glial populations in the parietal cortex of the rat. The study was performed in two groups of rats aged 4-6 and 30-32 months. Cortical volume, neuronal density, glial density, and neuronal area, and shapes of the soma and nucleus were analyzed in cortical layers I, II-IV, V, and VI using serial sections stained with cresyl-fast-violet, and quantitative morphometric techniques. No changes with age were found in volume of the cortex or neuronal density. Glial density increased significantly (mean for all layers 17%) in older rats. Layers II-IV, V, and VI showed an age-related decrease in the area of the neuronal soma. Neuronal shape, as revealed by the major/minor diameter ratio, also showed a decrease in old rats but only in layer II-IV. Nuclear area decreased with age only in layer VI. The stability of neuronal density together with the increased number of glial cells and the changes in neuronal soma size suggest that aged-related cognitive impairment could be a consequence of neuronal dysfunction rather than actual neuronal losses.