Impact of very old age on hypothalamic dopaminergic neurons in the female rat: a morphometric study

Dopaminergic neurons of the A(12) (tuberoinfundibular dopaminergic system) and A(14) (periventricular dopaminergic system) hypothalamic areas exert a tonic inhibitory control of prolactin secretion. Tuberoinfundibular dopaminergic system neuron function is known to decline during aging in rats, but little is known about the impact of extreme age on neuron number and morphology in the two systems. We morphometrically assessed the neurons of the tuberoinfundibular dopaminergic system and the periventricular dopaminergic system in female rats 6 (young, Y), 24 (old, O), and 30-32 (senescent, S) months old. Serial coronal sections of fixed hypothalami were immunohistochemically labeled for tyrosine hydroxylase, and immunoreactive perikarya from the A(12) and A(14) areas were quantitatively characterized and compared among the three age groups. Radioimmunoassay was used to measure serum prolactin. The number of A(12) tyrosine hydroxylase-immunoreactive perikarya showed a steady decline with age, whereas the number of A(14) tyrosine hydroxylase-immunoreactive perikarya remained stable from young to old age but showed a sharp drop in the senescent rats. In the old rats, tyrosine hydroxylase-immunoreactive neuronal area (A(12) = 135.37 and A(14) = 158.79 microm(2)) was significantly higher than that of young (A(12) = 72.56 and A(14) = 99.7 microm(2)) and senescent animals (A(12) = 95.5 and A(14) = 106.5 microm(2)). Densitometric assessment of median eminence tyrosine hydroxylase immunoreactivity revealed a steady age-related reduction of tyrosine hydroxylase content in the median eminence. Serum prolactin levels increased steadily with age. We conclude that, in the female rat, aging brings about a progressive loss of both tuberoinfundibular dopaminergic system and periventricular dopaminergic system neurons, which becomes more conspicuous at extreme ages.     

Gender and age differences in normal adult human brain: Voxel‐based morphometric study

The aim of this study was to evaluate the gender and age differences in the normal adult human brain, using voxel‐based morphometry. In this study, 227 right‐handed normal adults (male:female = 111:116) were examined. Three‐dimensional magnetic resonance brain images of each subject were segmented into gray and white matter using statistical parametric mappings. All individual data were transformed to standard brain space and then divided into older and younger age groups before examining the effects of age and gender. There was a significant negative correlation between gray matter concentration and age in each gender group. The differences were more prominent in the older age groups compared with the younger age groups. Gray matter concentrations in the bilateral inferior frontal lobes, anterior cingulate gyrus, medial thalamus, and hypothalamus were more retained in females as they aged, whereas those in the occipital regions were more retained in aging males. Our findings are consistent with biologically and hormonally established gender differences. Hum Brain Mapp, 2011. © 2010 Wiley‐Liss, Inc.     

Astroglial changes in the cerebral cortex of AIDS brains: a morphometric and immunohistochemical investigation

Astroglial changes in the cerebral cortex of AIDS brains were analysed by means of morphometry. Astrocytes with and without immunoreactivity for glial acidic protein (GFAP) were counted and their size was measured. In the two investigated cortical areas (frontal and parietal), a similar reaction pattern of astroglia was observed. The total number of astrocytes (i.e. GFAP-positive and GFAP-negative astrocytes) did not differ between control and AIDS brains. However, the number of GFAP-positive astrocytes was significantly increased in AIDS brains, while the number of GFAP-negative cells was significantly reduced. Nuclear size of GFAP-negative and GFAP-positive astrocytes was significantly increased. The reaction pattern of cortical astrocytes in AIDS seems to be characterized by GFAP production in protoplasmic astroglia as well as by hypertrophy of all astrocytes.     

A longitudinal study of age‐ and gender‐related annual rate of volume changes in regional gray matter in healthy adults

The aim of this study was to analyze correlations among the annual rate of gray matter volume change, age, gender, and cerebrovascular risk factors in 381 healthy community‐dwelling subjects with a large age range by applying a longitudinal design over 6 years using brain magnetic resonance images (MRIs). Brain MRI data were processed with voxel‐based morphometry using a custom template by applying diffeomorphic anatomical registration using the exponentiated lie algebra procedure. The annual rate of regional gray matter volume change showed significant positive correlations with age in several regions, including the bilateral temporal pole, caudate nucleus, ventral and dorsolateral prefrontal cortices, insula, hippocampus, and temporoparietal cortex, whereas significant negative correlations with age were observed in several regions including the bilateral cingulate gyri and anterior lobe of the cerebellum. Additionally, a significant age‐by‐gender interaction was found for the annual rate of regional gray matter volume change in the bilateral hippocampus. No significant correlations were observed between the annual rate of regional gray matter volume change and body mass index or systolic blood pressure. A significant positive correlation between the annual rate of gray matter volume change and age indicates that the region shows not linear but accelerated gray matter loss with age. Therefore, evaluating the annual rate of the gray matter volume change with age in healthy subjects is important in understanding how gray matter volume changes with aging in each brain region and in anticipating what cognitive functions are likely to show accelerated decline with aging. Hum Brain Mapp 34:2292–2301, 2013. © 2012 Wiley Periodicals, Inc.     

Three‐dimensional distribution of tyrosine hydroxylase,vasopressin and oxytocin neurones in the transparent postnatal mouse brain

Over the years, advances in immunohistochemistry techniques have been a critical step in detecting and mapping neuromodulatory substances in the central nervous system. The better quality and specificity of primary antibodies, new staining procedures and the spectacular development of imaging technologies have allowed such progress. Very recently, new methods permitting tissue transparency have been successfully used on brain tissues. In the present study, we combined whole‐mount immunostaining for tyrosine hydroxylase (TH), oxytocin (OXT) and arginine vasopressin (AVP), with the iDISCO+ clearing method, light‐sheet microscopy and semi‐automated counting of three‐dimensionally‐labelled neurones to obtain a (3D) distribution of these neuronal populations in a 5‐day postnatal (P5) mouse brain. Segmentation procedure and 3D reconstruction allowed us, with high resolution, to map TH staining of the various catecholaminergic cell groups and their ascending and descending fibre pathways. We show that TH pathways are present in the whole P5 mouse brain, similar to that observed in the adult rat brain. We also provide new information on the postnatal distribution of OXT and AVP immunoreactive cells in the mouse hypothalamus, and show that, compared to AVP neurones, OXT neurones in the supraoptic (SON) and paraventricular (PVN) nuclei are not yet mature in the early postnatal period. 3D semi‐automatic quantitative analysis of the PVN reveals that OXT cell bodies are more numerous than AVP neurones, although their immunoreactive soma have a volume half smaller. More AVP nerve fibres compared to OXT were observed in the PVN and the retrochiasmatic area. In conclusion, the results of the present study demonstrate the utility and the potency of imaging large brain tissues with clearing procedures coupled to novel 3D imaging technologies to study, localise and quantify neurotransmitter substances involved in brain and neuroendocrine functions.     

Response of cortical and pial arteries to changes of arterial CO2 tension in rats — a morphometric study

Changes of the internal (d) and external diameter (D) of cerebral arteries in response to the various levels of arterial pCO₂ were studied in anesthetized rats, of which brains were frozen in situ with isopentane cooled in liquid nitrogen. The parietal cortex was fixed with osmium tetroxide and stained with toluidine blue for morphometry of the cerebral arteries.In comparison with control animals (pCO₂ = 41.2mm Hg), the d/D ratio in animals with hypercapnia (pCO₂ = 69.9mm Hg) was increased by 11.7% in the pial arteries and 7.9% in the cortical arteries, indicating vasodilatation. In contrast, the d/D ratio in those with hypocapnia (pCO₂ = 27.7mm Hg) was decreased by 6.2% and 13.0%, respectively, indicating vasoconstriction. There was a significant linear correlation existing between the d/D ratio of either pial or cortical arteries and pCO₂. It is concluded that the cortical arteries respond to changes of arterial pCO₂ in a similar manner to the pial arteries.     

NIRS‐measured oxy‐ and deoxyhemoglobin changes associated with EEG spike‐and‐wave discharges in a genetic model of absence epilepsy: The GAERS

Purpose: Absence epilepsy may be severe and is frequently accompanied by cognitive delay, yet its metabolic/hemodynamic aspects have not been established. The Genetic Absence Epilepsy Rats from Strasbourg (GAERS) are an isomorphic, predictive, and homologous model of human absence epilepsy. We studied hemodynamic changes related to generalized spike‐and‐wave discharges (GSWDs) in GAERS by using a technique with high temporal resolution: near‐infrared spectroscopy (NIRS). We hypothesized that conflicting results from other techniques might be due to the averaging of a biphasic response such as the one we described in children. Methods: NIRS is particularly suitable for monitoring changes in the concentrations of oxy‐, deoxy‐, and total hemoglobin (HbO₂, HHb, and HbT), using the specific absorption properties of living tissues in the near infrared range. We obtained concomitant high quality electroencephalography (EEG)–NIRS recordings in six GAERS (total of 444 seizures), and tested whether the discharges were related to changes in cardiac or respiration rates. Results: The onset of GSWDs was preceded by a deactivation, followed by an activation that was possibly due to seizure‐suppression mechanisms. The end was marked by a deactivation. The onset of GSWDs was associated with a decrease and the end with a brief increase in respiratory rate. Discussion: Our results differ partially from those of previous studies on hemodynamic aspects of GSWDs (many of which describe a simple deactivation), probably due to differences in temporal resolution and data processing; however, they are consistent with metabolic studies, functional magnetic resonance imaging (fMRI) studies on WAG/Rij rats, and some results in children with absence epilepsy.     

Plaque‐dependent morphological and electrophysiological heterogeneity of microglia in an Alzheimer's disease mouse model

Microglia, the central nervous system resident innate immune cells, cluster around Aβ plaques in Alzheimer's disease (AD). The activation phenotype of these plaque‐associated microglial cells, and their differences to microglia distant to Aβ plaques, are incompletely understood. We used novel three‐dimensional cell analysis software to comprehensively analyze the morphological properties of microglia in the TgCRND8 mouse model of AD in spatial relation to Aβ plaques. We found strong morphological changes exclusively in plaque‐associated microglia, whereas plaque‐distant microglia showed only minor changes. In addition, patch‐clamp recordings of microglia in acute cerebral slices of TgCRND8 mice revealed increased K⁺ currents in plaque‐associated but not plaque‐distant microglia. Within the subgroup of plaque‐associated microglia, two different current profiles were detected. One subset of cells displayed only increased inward currents, while a second subset showed both increased inward and outward currents, implicating that the plaque microenvironment differentially impacts microglial ion channel expression. Using pharmacological channel blockers, multiplex single‐cell PCR analysis and RNA fluorescence in situ hybridization, we identified Kir and Kv channel types contributing to the in‐ and outward K⁺ conductance in plaque‐associated microglia. In summary, we have identified a previously unrecognized level of morphological and electrophysiological heterogeneity of microglia in relation to amyloid plaques, suggesting that microglia may display multiple activation states in AD.     

Three‐dimensional surface deformation‐based shape analysis of hippocampus and caudate nucleus in children with fetal alcohol spectrum disorders

Surface deformation‐based analysis was used to assess local shape variations in the hippocampi and caudate nuclei of children with fetal alcohol spectrum disorders. High‐resolution structural magnetic resonance imaging images were acquired for 31 children (19 controls and 12 children diagnosed with fetal alcohol syndrome/partial FAS). Hippocampi and caudate nuclei were manually segmented, and surface meshes were reconstructed. An iterative closest point algorithm was used to register the template of one control subject to all other shapes in order to capture the true geometry of the shape with a fixed number of landmark points. A point distribution model was used to quantify the shape variations in terms of a change in co‐ordinate positions. Using the localized Hotelling T² method, regions of significant shape variations between the control and exposed subjects were identified and mapped onto the mean shapes. Binary masks of hippocampi and caudate nuclei were generated from the segmented volumes of each brain. These were used to compute the volumes and for further statistical analysis. The Mann–Whitney test was performed to predict volume differences between the groups. Although the exposed and control subjects did not differ significantly in their volumes, the shape analysis showed the hippocampus to be more deformed at the head and tail regions in the alcohol‐exposed children. Between‐group differences in caudate nucleus morphology were dispersed across the tail and head regions. Correlation analysis showed associations between the degree of compression and the level of alcohol exposure. These findings demonstrate that shape analysis using three‐dimensional surface measures is sensitive to fetal alcohol exposure and provides additional information than volumetric measures alone. Hum Brain Mapp 35:659–672, 2014. © 2012 Wiley Periodicals, Inc.     

Regional gray matter reduction and theory of mind deficit in the early phase of schizophrenia: a voxel‐based morphometric study

Objective: We tested the association between theory of mind (ToM) performance and structural changes in the brains of patients in the early course of schizophrenia. Method: Voxel‐based morphometry (VBM) data of 18 patients with schizophrenia were compared with those of 21 controls. ToM skills were assessed by computerized faux pas (FP) tasks. Results: Patients with schizophrenia performed significantly worse in FP tasks than healthy subjects. VBM revealed significantly reduced gray matter density in certain frontal, temporal and subcortical regions in patients with schizophrenia. Poor FP performance of schizophrenics correlated with gray matter reduction in the left orbitofrontal cortex and right temporal pole. Conclusion: Our data indicate an association between poor ToM performance and regional gray matter reduction in the left orbitofrontal cortex and right temporal pole shortly after the onset of schizophrenia.     

Ketogenic diet ameliorates cognitive impairment and neuroinflammation in a mouse model of Alzheimer’s disease

IntroductionAlzheimer''s disease (AD) is the most common neurodegenerative disorder that causes dementia and affects millions of people worldwide. Although it has devastating outcomes for patients and tremendous economic costs to society, there is currently no effective treatment available.AimsThe high‐fat, low‐carbohydrate ketogenic diet (KD) is an established treatment for refractory epilepsy with a proven efficacy. Although the considerable interest has emerged in recent years for applying KD in AD patients, only few interventional studies in animals and humans have addressed the effects of KD on cognitive impairments, and the results were inconclusive. The aim of this study was to explore the impact of KD on cognitive functions and AD pathology in 5XFAD mice—a validated animal model of AD.ResultsFour months of a ketogenic diet improved spatial learning, spatial memory and working memory in 5XFAD mice. The improvement in cognitive functions was associated with a restored number of neurons and synapses in both the hippocampus and the cortex. Ketogenic diet treatment also reduced amyloid plaque deposition and microglial activation, resulting in reduced neuroinflammation. The positive effect of ketogenic diet on cognitive functions depended on the starting time and the duration of the diet. A shorter period (2 months) of ketogenic diet treatment had a weaker effect. Ketogenic diet initiated at late stage of AD (9 months of age) displayed no effect on cognitive improvement.ConclusionsThese findings indicate positive effects of ketogenic diet on both cognitive function and histopathology in Alzheimer''s disease, which could be due to reduced microglial activation and neuroinflammation. Our findings provide new insights and therapeutic interventions for the treatment of Alzheimer''s disease.     

Three‐dimensional structural changes in cerebral microvessels after transient focal cerebral ischemia in rats: Scanning electron microscopic study of corrosion casts

Pathological changes of cerebral microvessels in transient ischemia were investigated by scanning electron microscopy of vascular corrosion casts. Wistar rats were treated with middle cerebral artery (MCA) occlusion for 30 min, 1 h, 3 h, 4 h, 5 h or 7 h and subsequent reperfusion for 2 h. The ultrastructures of the cast were observed and computer‐aided montage micrographs were obtained for visualization of the whole microvasculature in the ischemic brain hemisphere. Avascular areas representing ischemic areas were detected in the frontotemporal cortex and caudate putamen in the groups from 30 min to 5 h occlusion. Extravasation of the resin, which probably corresponded to the leakage of plasma or hemorrhage, was seen as spheroidal, conglomerative, large massive and worm‐like types. The spheroidal type, which probably indicated a small leakage or minor hemorrhage, began to appear in the 30‐min occlusion group. The conglomerative type, which probably indicated a larger leakage or moderate hemorrhage, appeared in the 3‐ to 5‐h occlusion groups. The large massive and worm‐like types, which probably indicated a significant hemorrhage, appeared in the 4‐ and 5‐h occlusion groups. The number of these extravasations increased significantly in the 4‐h occlusion group. Arterioles near the avascular area frequently showed vasospastic appearances, such as corrugations, fusiform indentations of endothelial nuclei, continuous circulatory constrictions and severe narrowing with interrupted branches. Arteriolar vasospasm possibly caused prolonged hypoperfusion even if reperfusion was achieved. The capillaries had a thin stringy appearance in the 4‐ and 5‐h occlusion groups. These changes seemed to relate closely with increased intracranial pressure by brain edema or hemorrhage. The present study suggested that the risk of brain edema or hemorrhagic infarction increased beyond 3 h of MCA occlusion, and vasospasm of the arterioles might participate in stroke pathophysiology.     

Age‐related changes in cell dynamics of the postnatal mouse olfactory neuroepithelium: Cell proliferation,neuronal differentiation,and cell death

Age‐related changes in cell proliferation, neuronal differentiation, and cell death in mouse olfactory neuroepithelium were investigated. Mice at the age of 10 days through 16 months were given a single injection of bromodeoxyuridine (BrdU). The olfactory mucosae were fixed at 9 timepoints ranging from 2 hours to 3 months after the injection and examined using double immunostaining for BrdU and olfactory marker protein (OMP), and double staining with terminal deoxynucleotidyl transferase‐mediated biotinylated dUTP nick end labeling (TUNEL) and immunostaining for OMP. The number of BrdU‐labeled cells/mm epithelial length initially increased, peaked at 2–3 days after the BrdU injection, then declined at each age. The number of BrdU‐ and TUNEL‐labeled neuronal cells both decreased with increasing age, suggesting that the rates of both cell proliferation and cell death in the olfactory neuroepithelium decrease with increasing age. Double‐labeled cells for BrdU and OMP appeared at 7 days after injection in all age groups, suggesting that the time required for neuronal differentiation is broadly similar irrespective of age. In older age groups, smaller amounts of the newly produced cohort are integrated into the OMP‐positive ORN population, and even once it is integrated it is eliminated from the population more rapidly compared to the younger age groups. Furthermore, TUNEL assay showed that the fraction of apoptotic cells distributed in the OMP‐positive layer/total apoptotic cells decreased with age. This observation suggests that the turnover of mature ORNs is slower in the older neuroepithelium compared to the younger neuroepithelium. J. Comp. Neurol. 518:1962–1975, 2010. © 2010 Wiley‐Liss, Inc.     

Mid‐life environmental enrichment increases synaptic density in CA1 in a mouse model of Aβ‐associated pathology and positively influences synaptic and cognitive health in healthy ageing

Early‐life cognitive enrichment may reduce the risk of experiencing cognitive deterioration and dementia in later‐life. However, an intervention to prevent or delay dementia is likely to be taken up in mid to later‐life. Hence, we investigated the effects of environmental enrichment in wildtype mice and in a mouse model of Aβ neuropathology (APP_SWE/PS1_dE9) from 6 months of age. After 6 months of housing in standard laboratory cages, APP_SWE/PS1_dE9 (n = 27) and healthy wildtype (n = 21) mice were randomly assigned to either enriched or standard housing. At 12 months of age, wildtype mice showed altered synaptic protein levels and relatively superior cognitive performance afforded by environmental enrichment. Environmental enrichment was not associated with alterations to Aβ plaque pathology in the neocortex or hippocampus of APP_SWE/PS1_dE9 mice. However, a significant increase in synaptophysin immunolabeled puncta in the hippocampal subregion, CA1, in APP_SWE/PS1_dE9 mice was detected, with no significant synaptic density changes observed in CA3, or the Fr2 region of the prefrontal cortex. Moreover, a significant increase in hippocampal BDNF was detected in APP_SWE/PS1_dE9 mice exposed to EE, however, no changes were detected in neocortex or between Wt animals. These results demonstrate that mid to later‐life cognitive enrichment has the potential to promote synaptic and cognitive health in ageing, and to enhance compensatory capacity for synaptic connectivity in pathological ageing associated with Aβ deposition.     

Epidemiological patterns of tick‐borne encephalitis in Lithuania and clinical features in adults in the light of the high incidence in recent years: a retrospective study

Background and purpose

Lithuania is one of the countries with the highest incidence of tick‐borne encephalitis (TBE) in Europe. The aim of this study was to describe the epidemiological patterns of TBE in Lithuania, and characterize clinical features in adults in the light of the high incidence in recent years.

Methods

Surveillance data available on the website of the Centre for Communicable Diseases and AIDS of Lithuania were used to describe the epidemiological patterns of TBE. The retrospective study included 712 patients hospitalized in the Centre for Infectious Diseases and the Centre for Neurology of Vilnius University in the years 2005–2014.

Results

Tick‐borne encephalitis incidence rates have been increasing by 8.5% per year for the 45‐year period from 1970 to 2014. The joinpoint model finds two joinpoints at 1991 and 1994, with a significant decrease of 8.4% per year (P < 0.05) prior to the joinpoint at 1991, and a rise of 195.2% afterwards. TBE presented with meningoencephalitis in 556 cases (81.3%). A total of 129 patients (18%) had a severe case of the disease. The most common neurological signs were ataxia (579, 81.3%), meningeal signs (474, 66.5%) and tremor (338, 47.5%). Limb paresis was observed in 6.3% of patients. Five patients (0.7%) died, and 544 patients (76.7%) were discharged with sequelae.

Conclusions

Intensified efforts in promoting TBE vaccination will be needed in the light of the high incidence and expanded spatial distribution. Significant prognostic factors for severe cases of the disease were age above 61 and delayed immune response of specific immunoglobulin G.