Pathological Human Brain Research Articles

Selective gray matter staining of human brain slices: optimized use of cadaver materials

We report a novel staining technique for human brain slices that distinguishes clearly gray from white matter. Previously described techniques using either Prussian blue (Berlin blue) or phthalocyanine dyes usually have included a hot phenol pretreatment to prevent white matter staining. The technique we describe here does not require hot phenol pretreatment and allows the use of brains stored for postmortem periods of one to two years prior to staining. Our technique involves staining with copper(II) phthalocyanine-tetrasulfonic acid tetrasodium salt 1% in water for 2 h followed by acetic acid treatment; this produces excellent blue staining of gray matter with little white matter staining. The stained brain slices are excellent for teaching human brain anatomy and/or pathology, or for research purposes.

Movies of dopamine transporter occupancy with ultra-high resolution focusing pinhole SPECT

A pivotal question in neuropharmacology is how the function of neurotransmitter systems relates to psychiatric diseases. In experimental neuropharmacology, we have dreamt about a looking glass that would allow us to see neurotransmitter systems in action, and about animals that would faithfully serve us as models for human psychiatric disease. Analysis of animal models has been limited by the availability of methods to study in vivo neurotransmitter dynamics. Now, a single photon emission computed tomography system called U-SPECT can localize dopamine transporters in sub-compartments of the mouse brain during a range of points in time. Applied to the midbrain dopamine system of different models of disease, this will aid the understanding of dynamic processes of this neurotransmitter that underlie brain functions and human brain pathology.

Non‐invasive measurement of brain viscoelasticity using magnetic resonance elastography

The purpose of this work was to develop magnetic resonance elastography (MRE) for the fast and reproducible measurement of spatially averaged viscoelastic constants of living human brain. The technique was based on a phase-sensitive echo planar imaging acquisition. Motion encoding was orthogonal to the image plane and synchronized to intracranial shear vibrations at driving frequencies of 25 and 50 Hz induced by a head-rocker actuator. Ten time-resolved phase-difference wave images were recorded within 60 s and analyzed for shear stiffness and shear viscosity. Six healthy volunteers (six men; mean age 34.5 years; age range 25-44 years) underwent 23-39 follow-up MRE studies over a period of 6 months. Interindividual mean +/- SD shear moduli and shear viscosities were found to be 1.17 +/- 0.03 kPa and 3.1 +/- 0.4 Pas for 25 Hz and 1.56 +/- 0.07 kPa and 3.4 +/- 0.2 Pas for 50 Hz, respectively (P < or = 0.01). The intraindividual range of shear modulus data was 1.01-1.31 kPa (25 Hz) and 1.33-1.77 kPa (50 Hz). The observed modulus dispersion indicates a limited applicability of Voigt's model to explain viscoelastic behavior of brain parenchyma within the applied frequency range. The narrow distribution of data within small confidence intervals demonstrates excellent reproducibility of the experimental protocol. The results are necessary as reference data for future comparisons between healthy and pathological human brain viscoelastic data.

Localization of Parkinson’s disease-associated LRRK2 in normal and pathological human brain

Mutations in the LRRK2 gene cause autosomal dominant, late-onset parkinsonism, which presents with pleomorphic pathology including alpha-synucleopathy. To promote our understanding of the biological role of LRRK2 in the brain we examined the distribution of LRRK2 mRNA and protein in postmortem human brain tissue from normal and neuropathological subjects. In situ hybridization and immunohistochemical analysis demonstrate the expression and localization of LRRK2 to various neuronal populations in brain regions implicated in Parkinson’s disease (PD) including the cerebral cortex, caudate–putamen and substantia nigra pars compacta. Immunofluorescent double labeling studies additionally reveal the prominent localization of LRRK2 to cholinergic-, calretinin- and GABAB receptor 1-positive, dopamine-innervated, neuronal subtypes in the caudate–putamen. The distribution of LRRK2 in brain tissue from sporadic PD and dementia with Lewy bodies (DLB) subjects was also examined. In PD brains, LRRK2 immunoreactivity localized to nigral neuronal processes is dramatically reduced which reflects the disease-associated loss of dopaminergic neurons in this region. However, surviving nigral neurons occasionally exhibit LRRK2 immunostaining of the halo structure of Lewy bodies. Moreover, LRRK2 immunoreactivity is not associated with Lewy neurites or with cortical Lewy bodies in sporadic PD and DLB brains. These observations indicate that LRRK2 is not a primary component of Lewy bodies and does not co-localize with mature fibrillar alpha-synuclein to a significant extent. The localization of LRRK2 to key neuronal populations throughout the nigrostriatal dopaminergic pathway is consistent with the involvement of LRRK2 in the molecular pathogenesis of familial and sporadic parkinsonism.

Morphological and chemical studies of pathological human and mice brain at the subcellular level: Correlation between light, electron, and nanosims microscopies

Neurodegenerative diseases induce morphological and chemical alterations in well-characterized regions of the brain. Understanding their pathological processes requires the use of methods that assess both morphological and chemical alterations in the tissues. In the past, microprobe approaches such as scanning electron microscopy combined with an X-ray spectrometer, Proton induced X-ray emission, secondary ion mass spectrometry (SIMS), and laser microprobe mass analysis have been used for the study of pathological human brain with limited success. At the present, new SIMS instruments have been developed, such as the NanoSIMS-50 ion microprobe, that allow the simultaneous identification of five elements with high sensitivity, at subcellular spatial resolution (about 50-100 nm with the Cs(+) source and about 150-200 nm with O(-) source). Working in scanning mode, 2D distribution of five elements (elemental maps) can be obtained, thus providing their exact colocalization. The analysis can be performed on semithin or ultrathin embedded sections. The possibility of using transmission electron microscopy and SIMS on the same ultrathin sections allows the correlation between structural and analytical observations at subcellular and ultrastructural level to be established. Our observations on pathological brain areas allow us to establish that the NanoSIMS-50 ion microprobe is a highly useful instrument for the imaging of the morphological and chemical alterations that take place in these brain areas. In the human brain our results put forward the subcellular distribution of iron-ferritin-hemosiderin in the hippocampus of Alzheimer disease patients. In the thalamus of transgenic mice, our results have shown the presence of Ca-Fe mineralized amyloid deposits.

Electroencephalographic Intercentral Interaction as a Reflection of Normal and Pathological Human Brain Activity

The authors summarized EEG findings and defined the nature of the intercentral EEG relationships in different functional states in healthy subjects and patients with organic cerebral pathology, based on a coherence analysis. Similar EEG characteristics in healthy individuals were identified: an anterior-posterior gradient of average coherence levels, the type of cortical-subcortical relationships in anterior cerebral structures. Right- and left-handed individuals showed frequent and regional differences in EEG coherence, which mainly reflected specificity of intracortical relationships. Development and regression of pathology in right-and left-handed individuals with organic brain lesions were thought to be caused by these differences. Lesions of regulatory structures (diencephalic, brain stem and limbic structures) provoked a more diffused kind of changes of intercentral relationships, in contrast to cortical pathology. These changes tended to reciprocate. The dynamic nature of intercentral relationships and their interhemispheric differences was revealed when changing functional states of the brain (increase and decrease of functional level) in healthy individuals and patients with organic cerebral pathology in the process of conscious and psychic activity restoration. Changing activity predominance of certain regulatory structures was considered one of the most important factors determining the dynamic nature of EEG coherence.

Increased Generation of Neuronal Progenitors after Ischemic Injury in the Aged Adult Human Forebrain

The adult human brain retains the capacity to generate new neurons in the hippocampal formation (Eriksson et al., 1998) and neuronal progenitor cells (NPCs) in the forebrain (Bernier et al., 2000), but to what extent it is capable of reacting to injuries, such as ischemia, is not known. We analyzed postmortem tissue from normal and pathological human brain tissue (n = 54) to study the cellular response to ischemic injury in the forebrain. We observed that cells expressing the NPC marker polysialylated neural adhesion cell molecule (PSA-NCAM) are continuously generated in the adult human subventricular zone (SVZ) and migrate along the olfactory tracts. These cells were not organized in migrating chains as in the adult rodent rostral migratory stream, and their number was lower in the olfactory tracts of brains from old (56-81 years of age) compared with young (29 + 36 years of age) individuals. Moreover, we show that in brains of patients of advanced age (60-87 years of age), ischemia led to an elevated number of Ki-67-positive cells in the ipsilateral SVZ without concomitant apoptotic cell death. Additionally, ischemia led to an increased number of PSA-NCAM-positive NPCs close to the lateral ventricular walls, compared with brains of comparable age without obvious neuropathologic changes. These results suggest that the adult human brain retains a capacity to respond to ischemic injuries and that this capacity is maintained even in old age.

LRRK2 Expression in Normal and Pathologic Human Brain and in Human Cell Lines

Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) have been recently identified in families with autosomal-dominant late-onset Parkinson disease. We report that by reverse transcriptase-polymerase chain reaction, the mRNA of LRRK2 is expressed in soluble extracts of human brain, liver, and heart and in cultured human astrocytes, microglia, and oligodendroglia as well as in human neuroblastoma cell lines. We find by Western blotting using a polyclonal antibody of the leucine-rich repeat kinase 2 protein (Lrrk2) specific for C-terminal residues 2511-2527 that an apparent full-length protein and several of its fractions are expressed in soluble extracts of normal human brain. By immunocytochemistry, the antibody recognizes neurons, and more weakly astrocytes and microglia, in normal brain tissue. It intensely labels Lewy bodies in Parkinson disease and related neurodegenerative disorders. It also labels a subset of neurofibrillary tangles in Alzheimer disease and the Parkinsonism dementia complex of Guam (PDCG). It labels thorn-shaped astrocytes and oligodendroglial coiled bodies in PDCG; oligodendroglial inclusions in multiple system atrophy; Pick bodies in Pick disease; nuclear and cytoplasmic inclusions in Huntington disease; and intraneuronal and glial inclusions in amyotrophic lateral sclerosis. In summary, LRRK2 is constitutively expressed in neurons and also in glial cells of human brain. It strongly associates with pathological inclusions in several neurodegenerative disorders.

A low temperature embedding and section registration strategy for 3D image reconstruction of the rat brain from autoradiographic sections

In studies on animal models of human brain pathologies, three-dimensional reconstruction from histological sections is particularly useful when assessing the morphologic, functional and biochemical changes induced by pathology. It allows assessing lesion heterogeneity in planes different from the cutting plane and allows correlating the histology with images obtained in vivo, such as by means of magnetic resonance imaging. To create a 3D volume from autoradiographic sections with minimal distortion, both cryosectioning as well as section registration need to be optimal. This paper describes a strategy whereby four external fiducial markers are positioned outside the rat brain with the use of a low temperature brain embedding procedure. The fiducial markers proposed here can be rapidly added to any frozen tissue block with no impact on the subsequent histological operations. Since embedding is performed at a low temperature, no tissue degradation occurs due to sample heating. The markers enable robust and almost error free registration, even in the presence of missing sections and poor image quality. Furthermore, the markers may be used to partially correct for global distortions.

Longitudinal Brain Magnetic Resonance Imaging Study of the Alcohol‐Preferring Rat. Part II: Effects of Voluntary Chronic Alcohol Consumption

Tracking the dynamic course of human alcoholism brain pathology can be accomplished only through naturalistic study and without opportunity for experimental manipulation. Development of an animal model of alcohol-induced brain damage, in which animals consume large amounts of alcohol following cycles of alcohol access and deprivation and are examined regularly with neuroimaging methods, would enable hypothesis testing focused on the degree, nature, and factors resulting in alcohol-induced brain damage and the prospects for recovery or relapse. We report the results of longitudinal magnetic resonance imaging (MRI) studies of the effects of free-choice chronic alcohol intake on the brains of 2 cohorts of selectively bred alcohol-preferring (P) rats. In the companion paper, we described the MRI acquisition and analysis methods, delineation of brain regions, and growth patterns in total brain and selective structures of the control rats in the present study. Both cohorts were studied as adults for about 1 year and consumed high doses of alcohol for most of the study duration. The paradigm involved a 3-bottle choice with 0, 15 (or 20%), and 30% (or 40%) alcohol available in several different exposure schemes: continuous exposure, cycles of 2 weeks on followed by 2 weeks off alcohol, and binge drinking in the dark. Brain structures of the adult P rats in both the alcohol-exposed and the water control conditions showed significant growth, which was attenuated in a few measures in the alcohol-exposed groups. The region with the greatest demonstrable effect was the corpus callosum, measured on midsagittal images. The P rats showed an age-alcohol interaction different from humans, in that normal growth in selective brain regions that continues in adult rats was retarded.

Microvascular pathology in the aging human brain: Evidence that senile plaques are sites of microhaemorrhages

Amyloid-rich plaques are a feature of the aging human cerebral cortex. We have recently described another feature of aging human cortex, microhaemorrhages, identified by their content of haem, red blood cells, collagen and clotting factors, and their spatial relationship to capillaries. Here we relate microhaemorrhages to amyloid deposits. Observations were made in three groups: patients with no history of dementia, patients with Alzheimer's disease (AD) and patients with Down's syndrome (DS) and dementia. Amyloid deposits and microhaemorrhages were labelled in adjacent sections, amyloid deposits with antibodies to β-amyloid (βA), and microhaemorrhages by Prussian blue histochemistry for haem. The densities and sizes of βA deposits and haem-rich deposits (HRDs), and their relationship to blood vessels, were surveyed in temporal, cingulate and superior frontal cortex. Our results suggest that HRDs and βA deposits are the same sites of pathology. Their densities in the cortex and white matter of the regions surveyed varied markedly between cases, particularly between demented and non-demented cases, but they always co-varied; where haem deposits were sparse or numerous, so were βA deposits. Both HRDs and βA deposits formed adjacent to or encircling small vessels, often at branch points, and a spatial proximity analysis confirmed that both were found close to or colocalising with microvessels. Both HRDs and βA deposits were associated with blood- or vessel-derived proteins (fibrinogen, von Willebrand factor and collagen VI). Since haem is an established marker of cerebral bleeding, and amyloid is a marker of senile plaques, our results indicate that senile plaques are sites of microhaemorrhages. This colocalisation raises the very testable questions of whether microhaemorrhages are early events in plaque formation and whether therapies which stabilise cerebral microvessels can prevent the onset or slow the progress of dementias associated with plaque formation.

Similar calcification process in acute and chronic human brain pathologies

Cellular microcalcification observed in a diversity of human pathologies, such as vascular dementia, Alzheimer's disease, Parkinson's disease, astrogliomas, and posttraumatic epilepsy, also develops in rodent experimental models of central nervous system (CNS) neurodegeneration. Central to the neurodegenerative process is the inability of neurons to regulate intracellular calcium levels properly, and this is extensible to fine regulation of the CNS. This study provides evidence of a common pattern of brain calcification taking place in several human pathologies, and in the rat with glutamate-derived CNS lesions, regarding the chemical composition, physical characteristics, and histological environment of the precipitates. Furthermore, a common physical mechanism of deposit formation through nucleation, lineal growth, and aggregation is presented, under the modulation of protein deposition and elemental composition factors. Insofar as calcium precipitation reduces activity signals at no energy expense, the presence in human and rodent cerebral brain lesions of a common pattern of calcification may reflect an imbalance between cellular signals of activity and energy availability for its execution. If this is true, this new step of calcium homeostasis can be viewed as a general cellular adaptative mechanism to reduce further brain damage.

Imaging ex vivo healthy and pathological human brain tissue with ultra-high-resolution optical coherence tomography

The ability of ultra-high-resolution optical coherence tomography (UHR OCT) to discriminate between healthy and pathological human brain tissue is examined by imaging ex vivo tissue morphology of various brain biopsies. Micrometer-scale OCT resolution (0.9x2 microm, axialxlateral) is achieved in biological tissue by interfacing a state-of-the-art Ti:Al2O3 laser (lambda(c)=800 nm, delta lambda=260 nm, and P(out)=120 mW exfiber) to a free-space OCT system utilizing dynamic focusing. UHR OCT images are acquired from both healthy brain tissue and various types of brain tumors including fibrous, athypical, and transitional meningioma and ganglioglioma. A comparison of the tomograms with standard hematoxylin and eosin (H&E) stained histological sections of the imaged biopsies demonstrates the ability of UHR OCT to visualize and identify morphological features such as microcalcifications (>20 microm), enlarged nuclei of tumor cells (approximately 8 to 15 microm), small cysts, and blood vessels, which are characteristic of neuropathologies and normally absent in healthy brain tissue.

TPPP/p25: from unfolded protein to misfolding disease: prediction and experiments

TPPP/p25, the first representative of a new protein family, identified as a brain-specific unfolded protein induces aberrant microtubule assemblies in vitro, suppresses mitosis in Drosophila embryo and is accumulated in inclusion bodies of human pathological brain tissues. In this paper, we present prediction and additional experimental data that validate TPPP/p25 to be a new member of the “intrinsically unstructured” protein family. The comparison of these characteristics with that of α-synuclein and tau, involved also in neurodegenerative diseases, suggested that although the primary sequences of these proteins are entirely different, there are similarities in their well-defined unstructured segments interrupted by “stabilization centres”, phosphorylation and tubulin binding motives. SK-N-MC neuroblastoma cells were transfected with pEGFP-TPPP/p25 construct and a stable clone denoted K4 was selected and used to establish the effect of this unstructured protein on the energy state/metabolism of the cells. Our data by analyzing the mitochondrial membrane polarization by fluorescence microscopy revealed that the high-energy phosphate production in K4 clone is not damaged by the TPPP/p25 expression. Biochemical analysis with cell homogenates provided quantitative data that the ATP level increased 1.5-fold and the activities of hexokinase, glucosephosphate isomerase, phosphofructokinase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase were 1.2 to 2.0-fold higher in K4 as compared to the control. Our modelling using these data and rate equations of the individual enzymes suggests that the TPPP/p25 expression stimulates glucose metabolism. At pathological conditions TPPP/p25 is localized in inclusion bodies in multiple system atrophy, it tightly co-localizes with α-synuclein, partially with tubulin and not with vimentin. The previous and the present studies obtained with immunohistochemistry with pathological human brain tissues rendered it possible to classify among pathological inclusions on the basis of immunolabelling of TPPP/p25, and suggest this protein to be a potential linkage between Parkinson’s and Alzheimer’s diseases.

Electron nanodiffraction and high-resolution electron microscopy studies of the structure and composition of physiological and pathological ferritin

Structures of core nanocrystals of physiological (horse spleen, human liver, and brain) and pathological human brain of patients with progressive supranuclear palsy (PSP) and Alzheimer’s disease (AD) ferritin molecules were determined using electron nanodiffraction and high-resolution transmission electron microscopy. The poly-phasic structure of the ferritin cores is confirmed. There are significant differences in the mineral composition between the physiological and pathological ferritins. The physiological ferritin cores mainly consist of single nanocrystals containing hexagonal ferrihydrite (Fh) and hematite (Hm) and some cubic magnetite/maghemite phase. In the pathological cores, Fh is present but only as a minor phase and Hm is absent. The major phases are a face-centered-cubic (fcc) structure with a=0.43 nm and a high degree of disorder, related to wustite, and a cubic magnetite-like structure. These two cubic phases are also present in human aged normal brain. Evidence for the presence of hemosiderin together with ferritin in the pathological brains is deduced from the similarities of the diffraction patterns with those from patients with primary hemochromatosis, and differences in the shapes and protein composition of the protein shell. These findings suggest a disfunction of the ferritin associated with PSP and AD, associated with an increase in the concentration of brain ferrous toxic iron.

MRI texture analysis on texture test objects, normal brain and intracranial tumors.

Texture analysis was performed in three different MRI units on T1 and T2-weighted MR images from 10 healthy volunteers and 63 patients with histologically confirmed intracranial tumors. The goal of this study was a multicenter evaluation of the usefulness of this quantitative approach for the characterization of healthy and pathologic human brain tissues (white matter, gray matter, cerebrospinal fluid, tumors and edema). Each selected brain region of interest was characterized with both its mean gray level values and several texture parameters. Multivariate statistical analyses were then applied in order to discriminate each brain tissue type represented by its own set of texture parameters. Texture analysis was previously performed on test objects to evaluate the method dependence on acquisition parameters and consequently the interest of a multicenter evaluation. Even obtained on different sites with their own acquisition routine protocol, MR brain images contain textural features that can reveal discriminant factors for tissue classification and image segmentation. It can also offer additional information in case of undetermined diagnosis or to develop a more accurate tumor grading.

Brain plasticity and microglia: is transsynaptic glial activation in the thalamus after limb denervation linked to cortical plasticity and central sensitisation?

Microglia are a subset of tissue-macrophages that are ubiquitously distributed throughout the entire CNS. In health, they remain largely dormant until activated by a pathological stimulus. The availability of more sensitive detection techniques has allowed the early measurement of the cell responses of microglia in areas with few signs of active pathology. Subtle neuronal injury can induce microglial activation in retrograde and anterograde projection areas remote from the primary lesion focus. There is also evidence that in cases of long-standing abnormal neuronal activity, such as in patients after limb amputation with chronic pain and phantom sensations, glial activation may occur transsynaptically in the thalamus. Such neuronally driven glial responses may be related to the emergence central sensitisation in chronic pain states or plasticity phenomena in the cerebral cortex. It is suggested, that such persistent low-level microglial activation is not adequately described by the traditional concept of phagocyte-mediated tissue damage that largely evolved from studies of acute brain lesion models or acute human brain pathology. Due to the presence of signal molecules that can act on neurons and microglia alike, the communication between neurons and microglia is likely to be bi-directional. Persistent subtle microglial activity may modulate basal synaptic transmission and thus neuronal functioning either directly or through the interaction with astrocytes. The activation of microglia leads to the emergence of microstructural as well as functional compartments in which neurokines, interleukins and other signalling molecules introduce a qualitatively different, more open mode of cell–cell communication that is normally absent from the healthy adult brain. This ‘neo-compartmentalisation’, however, occurs along predictable neuronal pathways within which these glial changes are themselves under the modulatory influence of neurons or other glial cells and are subject to the evolving state of the pathology. Depending on the disease state, yet relatively independent of the specific disease cause, fluctuations in the modulatory influence by non-neuronal cells may form the cellular basis for the variability of brain plasticity phenomena, i.e. the plasticity of plasticity.

Pulsed Z-spectroscopic imaging of cross-relaxation parameters in tissues for human MRI: theory and clinical applications.

A new method of pulsed Z-spectroscopic imaging is proposed for in vivo visualization and quantification of the parameters describing cross-relaxation between protons with liquid-like and solid-like relaxation properties in tissues. The method is based on analysis of the magnetization transfer (MT) effect as a function of the offset frequency and amplitude of a pulsed off- resonance saturation incorporated in a spoiled gradient-echo MRI pulse sequence. The theoretical concept of the method relies on an approximated analytical model of pulsed MT that provides a simple three-parameter equation for a pulsed steady-state Z-spectrum taken far from resonance. Using this model, the parametric images of cross-relaxation rate constant, content, and T(2) of the semisolid proton fraction can be reconstructed from a series of MT-weighted images and a coregistered T(1) map. The method was implemented on a 0.5 T clinical MRI scanner, and it provided high-quality 3D parametric maps within an acceptable scanning time. The estimates of cross-relaxation parameters in brain tissues were shown to be quantitatively consistent with the literature data. Clinical examples of the parametric images of human brain pathologies (multiple sclerosis and glioma) demonstrated high tissue contrast and clear visualization of the lesions.

Reduced degree of long-range phase synchrony in pathological human brain

In this paper multivariate spontaneous EEG signals from three broad groups of human subjects - control, seizure, and mania - were studied with the aim of investigating the possible effect of these pathologies on the degree of phase synchronization between cortical areas. The degree of phase synchrony was measured by two recently developed measures which are more suitable than classical indices like correlation or coherence when dealing with nonlinear and non-stationary signals like the EEG. Signals were reduced to seven frequency bands (delta, theta, alpha-1, alpha-2, beta-1, beta-2 and gamma) which were statistically compared between the normal and the other two groups. It was found that the degree of long-range synchrony was significantly reduced for both pathological groups as compared with the control group. No clear differences were found in the degrees of short-range synchrony.

Morphometry of E-PTA stained synapses at the periphery of pathological lesions

We carried out a novel application of the disector sampling and counting method, in a biopsy material from the pathologic human brain, to estimate the synaptic structural dynamics, quantitatively. Parietal cortex biopsies of adult (mean age: 40.0 years) and old (mean age: 66.2 years) patients having undergone surgical intervention were investigated. The tissue samples were excised at the periphery of meningioma masses. Synaptic contact zones were stained en bloc by the ethanol phosphotungstic acid (E-PTA) preferential technique which selectively enhances both the pre- and post-synaptic paramembranous material separated by a sharp cleft against a very faint background, thus facilitating and objectifying synaptic morphometry. The disector method, associated with currently used morphometric formulas, enabled us to measure the number of synapses/μm3 of tissue (numeric density: Nv); the total area of the synaptic contact zones/μm3 of tissue (surface density: Sv) and the average synaptic size (S). In old vs. adult patients, Nv decreased by 7.5% (Mean (SEM): Adult 2.0040(0.0452); Old 1.6780(0.0623)), while S increased by 17.5% (Adult 0.0203(0.0026); Old 0.0246(0.0035)). Sv did not show any age-related difference. The same negative correlation between Nv and S has also been reported in physiological aging, and this suggests the active presence of age-related synaptic restructuring mechanisms in the nervous tissue surrounding a tumoral mass.