Density Of Neurites Research Articles

Understanding microstructure of the brain by comparison of neurite orientation dispersion and density imaging (NODDI) with transparent mouse brain

BackgroundNeurite orientation dispersion and density imaging (NODDI) is a diffusion magnetic resonance imaging (MRI) technique with the potential to visualize the microstructure of the brain. Revolutionary histological methods to render the mouse brain transparent have recently been developed, but verification of NODDI by these methods has not been reported.PurposeTo confirm the concordance of NODDI with histology in terms of density and orientation dispersion of neurites of the brain.Material and MethodsWhole brain diffusion MRI of a thy-1 yellow fluorescent protein mouse was acquired with a 7-T MRI scanner, after which transparent brain sections were created from the same mouse. NODDI parameters calculated from the MR images, including the intracellular volume fraction (Vic) and the orientation dispersion index (ODI), were compared with histological findings. Neurite density, Vic, and ODI were compared between areas of the anterior commissure and the hippocampus containing crossing fibers (crossing areas) and parallel fibers (parallel areas), and the correlation between fiber density and Vic was assessed.ResultsThe ODI was significantly higher in the crossing area compared to the parallel area in both the anterior commissure and the hippocampus (P = 0.0247, P = 0.00022, respectively). Neurite density showed a similar tendency, but was significantly different only in the hippocampus (P = 7.91E−07). There was no significant correlation between neurite density and Vic.ConclusionNODDI was verified by histology for quantification of the orientation dispersion of neurites. These results indicate that the ODI is a suitable index for understanding the microstructure of the brain in vivo.

Ketamine Promotes Neurite Growth and Recovers Dexamethasone-Induced Mobility Decrease in Cultured Hippocampal Cells

The “plasticity hypothesis” proposes that major depression is caused by morphological and biochemical modifications in neurons and astrocytes and that the illness is at least partially associated with modifications of cellular communications between these cells. In this study we examined effects of Ketamine, an NMDA antagonist that was conventionally used as an anesthetic, on dexamethasone-treated neurons. Several clinical trials have shown that this drug has very potent anti-depressive properties, becoming effective within a few hours. Ketamine has also been shown to have fast effect on the neuronal plasticity, including synaptogenesis and dendrite formation. In this project, we studied its morphodynamic effects in hippocampal neuronal cultures by motion-, neurite growth analysis and cluster formation. Time-lapse images taken over 48 hours and the resulting videos were analyzed by several newly developed complex algorithms for morphometry and cell mobility. Dexamethasone, a synthetic corticosteroid, was used to induce depression-associated changes in cultured hippocampal neurons. Results show that ketamine significantly neutralized dexamethasone-induced reduction of cell mobility and cell cluster formation. Neither dexamethasone nor ketamine had strong effects on the density and growth of neurites. In summary, given the complex actions of ketamine on morphodynamic parameters in cultured neurons, its mechanism as an anti-depressant cannot be explained with its NMDA-antagonistic properties alone but implies additional impact on the formation of communication networks and morphology related pathways.

Design of Hyaluronic Acid Hydrogels to Promote Neurite Outgrowth in Three Dimensions.

A hyaluronic acid (HA)-based extracellular matrix (ECM) platform with independently tunable stiffness and density of cell-adhesive peptide (RGD, arginine-glycine-aspartic acid) that mimics key biochemical and mechanical features of brain matrix has been designed. We demonstrated here its utility in elucidating ECM regulation of neural progenitor cell behavior and neurite outgrowth. The analysis of neurite outgrowth in 3-D by two-photon microscopy showed several important results in the development of these hydrogels. First, the ability of neurites to extend deeply into these soft HA-based matrices even in the absence of cell-adhesive ligand further confirms the potential of HA hydrogels for central nervous system (CNS) regeneration. Second, the behavior of hippocampal neural progenitor cells differed markedly between the hydrogels with a storage modulus of 400 Pa and those with a modulus of 800 Pa. We observed an increased outgrowth and density of neurites in the softest hydrogels (G' = 400 Pa). Interestingly, cells seeded on the surface of the hydrogels functionalized with the RGD ligand experienced an optimum in neurite outgrowth as a function of ligand density. Surprinsingly, neurites preferentially progressed inside the gels in a vertical direction, suggesting that outgrowth is directed by the hydrogel structure. This work may provide design principles for the development of hydrogels to facilitate neuronal regeneration in the adult brain.

Changes in the sympathetic innervation of the gut in rotenone treated mice as possible early biomarker for Parkinson's disease.

IntroductionInvolvement of the peripheral nervous system (PNS) is relatively common in Parkinson’s disease (PD) patients. PNS alterations appear early in the course of the disease and are responsible for some of the non-motor symptoms observed in PD patients. In previous studies, we have shown that environmental toxins can trigger the disease by acting on the enteric nervous system.Material and methods Here, we analyzed the effect of mitochondrial Complex I inhibition on sympathetic neuritis in vivo and sympathetic neurons in vitro. Combining in vivo imaging and protein expression profiling.Results we found that rotenone, a widely used mitochondrial Complex I inhibitor decreases the density of sympathetic neurites innervating the gut in vivo, while in vitro, it induces the redistribution of intracellular alpha-synuclein and neurite degeneration. Interestingly, sympathetic neurons are much more resistant to rotenone exposure than mesencephalic dopaminergic neurons.ConclusionAltogether, these results suggest that enteric sympathetic denervation could be an initial pre-motor alteration in PD progression that could be used as an early biomarker of the disease.Electronic supplementary materialThe online version of this article (doi:10.1007/s10286-016-0358-6) contains supplementary material, which is available to authorized users.

Retrograde Melanopsin Signaling Increases With Age in Retinal Degenerate Mice Lacking Rods and the Majority of Cones.

Following on from reports of retrograde retinal signaling in mice, we sought to investigate the influence of age and retinal location on this phenomenon using mice that lack rods and the majority of cones. We used functional anatomy for c-fos (Fos) and tyrosine hydroxylase (TH) to measure light-driven activation of dopamine neurons along a dorsal-ventral transect in C3H/He wild-type and rodless-coneless rd/rd cl (rdcl) mice aged 3, 5, and >14 months. A parallel series of retinae from 3-month-old mice was also stained for cone opsins and melanopsin. Analysis by confocal microscopy revealed light-driven Fos activation in TH cells residing in the middorsal retina of the youngest rdcl mice. This region was largely devoid of residual cones but contained a large number of intrinsically photosensitive retinal ganglion cells (ipRGCs) and the highest density of melanopsin neurites. With advancing age, there was a paradoxical increase in retrograde signaling from ∼3% Fos-positive (Fos+) TH cells at 3 months to ∼36% in rdcl mice >14 months. This increased activation occurred in more central and peripheral retinal regions. Our data provide new insights into the anatomy and plasticity of retrograde melanopsin signaling in mice with severe rod/cone dystrophy. The increased retrograde signaling we detect may result from either an increased potency of melanopsin signaling with advancing age and/or postsynaptic modification to dopaminergic neurons.

White matter microstructure pathology in classic galactosemia revealed by neurite orientation dispersion and density imaging.

White matter abnormalities have been observed in patients with classic galactosemia, an inborn error of galactose metabolism. However, magnetic resonance imaging (MRI) data collected in the past were generally qualitative in nature. Our objective was to investigate white matter microstructure pathology and examine correlations with outcome and behaviour in this disease, by using multi-shell diffusion weighted imaging. In addition to standard diffusion tensor imaging (DTI), neurite orientation dispersion and density imaging (NODDI) was used to estimate density and orientation dispersion of neurites in a group of eight patients (aged 16–21 years) and eight healthy controls (aged 15–20 years). Extensive white matter abnormalities were found: neurite density index (NDI) was lower in the patient group in bilateral anterior areas, and orientation dispersion index (ODI) was increased mainly in the left hemisphere. These specific regional profiles are in agreement with the cognitive profile observed in galactosemia, showing higher order cognitive impairments, and language and motor impairments, respectively. Less favourable white matter properties correlated positively with age and age at onset of diet, and negatively with behavioural outcome (e.g. visual working memory). To conclude, this study provides evidence of white matter pathology regarding density and dispersion of neurites in these patients. The results are discussed in light of suggested pathophysiological mechanisms.

Neurites outgrowth and amino acids levels in goldfish retina under hypo-osmotic or hyper-osmotic conditions

Amino acids are known to play relevant roles as osmolytes in various tissues, including the retina. Taurine is one of these active molecules. In addition, taurine stimulates outgrowth from the goldfish retina by mechanisms that include extracellular matrix, calcium fluxes and protein phosphorylation. The present report aims to explore the effect of medium osmolarity on goldfish retinal outgrowth and the possible modifications produced by changing eye osmolarity on amino acid levels in the retina. Goldfish retinal explants were obtained 10 days after crush of the optic nerve and cultured under iso-, hypo- or hyper-osmotic conditions. Hypo-osmotic medium was prepared by diluting the solutions 10% twice, preserving fetal calf serum concentration. Hyper-osmotic medium was done by adding 50 or 100 mM urea or mannitol. Evaluation of length and density of neurites was performed 5 days after plating. Outgrowth was reduced in hypo- and in hyper-osmotic conditions. Taurine, 4 mM, increased length and density of neurites in iso-osmotic, and produced stimulatory effects under both hyper-osmotic conditions. The in vivo modification of osmolarity by intraocular injection of water or 100 mM urea modified levels of free amino acids in the retina. Taurine and aspartate retinal levels increased in a time-dependent manner after hypo- and hyper-osmotic solution injections. Serine, threonine, arginine, γ-aminobutyric acid, alanine and tyrosine were elevated in hyper-osmotic conditions. Outgrowth in vitro, after in vivo osmolarity changes, was higher in the absence of taurine, but did not increase in the presence of the amino acid. The fact that certain outgrowth took place in these conditions support that the impairment was not due to tissue damage. Rather, the effects might be related to the cascade of kinase events described during osmolarity variations. The time course under these conditions produced adjustments in ganglion cells probably related to taurine transporter, and phosphorylation of specific proteins.

Cytoskeletal alterations differentiate presenilin-1 and sporadic Alzheimer’s disease

Most cases of Alzheimer's disease (AD) are sporadic in nature, although rarer familial AD (FAD) cases have provided important insights into major pathological disease mechanisms. Mutations in the presenilin 1 gene (PS1) are responsible for the majority of FAD cases, causing an earlier age of onset and more rapid progression to end-stage disease than seen in sporadic AD. We have investigated the cytoskeletal alterations in neuritic AD pathology in a cohort of FAD cases in comparison to sporadic AD and pathologically aged cases. Tau-immunoreactive neurofibrillary tangle (NFT) loads were similar between PS1 FAD and sporadic AD cases. Similarly, plaque loads, both beta-amyloid (Abeta) and thioflavine S, in PS1 FAD and sporadic AD cases were not significantly different; however, in pathologically aged cases, they were significantly lower than those in PS1 cases, but were not different from sporadic AD cases. The 'cotton wool' plaque characteristic of PS1 cases did not demonstrate a high density of dystrophic neurites compared to other Abeta plaque types, but did demonstrate a localised mass effect on the neuropil. Despite minimal differences in plaque and NFT loads, immunolabelling demonstrated clear phenotypic differences in the NFTs and dystrophic neurites in PS1 FAD cases. Presenilin-1 cases exhibited significantly (P < 0.05) more tau-positive NFTs that were immunolabelled by the antibody SMI312 (anti-phosphorylated NF protein and phosphorylated tau) than sporadic AD cases. Presenilin-1 cases also exhibited numerous ring-like NF-positive and elongated tau-labelled dystrophic neurites, whereas these dystrophic neurite types were only abundant at the very early (pathologically aged cases) or very late stages of sporadic AD progression, respectively. These differences in cytoskeletal pathology in PS1 cases suggest an accelerated rate of neuritic pathology development, potentially due to mutant PS1 influencing multiple pathogenic pathways.

C‐Type Natriuretic Peptide is a Schwann Cell‐Derived Factor For Development and Function of Sensory Neurones

Cyclic GMP (cGMP) is known to play important roles for neuronal development and neurite pathfinding. However, the regulatory mechanism that governs the synthesis of cGMP in the nervous system is not well defined. In the present study, we examined the role of C-type natriuretic peptide (CNP), which increases intracellular cGMP upon binding to its receptor, guanylyl cyclase (GC)-B, in the peripheral nervous system. Immunohistochemistry revealed that CNP is demonstrated in Schwann cells, whereas GC-B mRNA is highly expressed in dorsal root ganglion (DRG) neurones. In cultured DRG neurones, GC-B was demonstrated in dendrites of TrkA-positive cells, where it co-exists with cGMP-dependent protein kinase I (cGKI), the major intracellular mediator of cGMP actions. Addition of CNP in the culture medium increased the density of fine neurites, which was accompanied by the increase in phosphorylation of vasodilator-stimulated phosphoprotein, a cGKI substrate. Furthermore, in mice deficient for the CNP gene (CNP-KO), the numbers of TrkA-positive DRG neurones were diminished. Likewise, there were much less cGKI-positive neurones in DRG and cGKI-positive fibres in the dorsal spinal cord of CNP-KO than wild-type mice. Finally, the bone deformity-rescued CNP-KO mice displayed a decreased response to formalin-induced pain compared to wild-type. Taken together, these results suggest that CNP is derived from Schwann cells and plays an important role for the development and function of nociceptive sensory neurones.

Increased α1D adrenergic receptor activity and protein expression in the urinary bladder of aged rats

A possibility that aging affects (a) expression of the alpha1D-adrenergic receptor (AR1D), (b) AR1D-mediated contractions and (c) sympathetic innervation in the urinary bladder in rats was studied. Contraction produced by phenylephrine and inhibition of these contractions by a non-selective alpha1-adrenergic antagonist prazosin and a selective AR1D antagonist BMY7378 were compared between 6- and 24-month-old Fisher rats. Expressions of VMAT and AR1D in the bladder were assessed by immunofluorescence and Western blot. Phenylephrine-induced contractions were larger and inhibition of these contractions by BMY7378 was significantly greater in 24-month-old rats. Aging increased expression of AR1D in the bladder. Density of VMAT-immunoreactive neurites was decreased in smooth muscle but elevated in the suburothelial region of 24-month-old rats. The results suggest that influence of adrenergic activity on bladder contractility increases with aging is due to overexpression of the AR1D. Influence of adrenergic activity on the urothelial function may also be enhanced with aging.

Cardiac sympathetic denervation correlates with clinical and pathologic stages of Parkinson's disease

Attention has been drawn to cardiac sympathetic denervation in Parkinson's disease (PD) based on clinical studies using [123I] metaiodobenzylguanidine scintigraphy; however, the histologic correlates and time course of cardiac sympathetic denervation are poorly understood. To address these issues, we used tyrosine hydroxylase (TH) immunohistochemistry to detect cardiac sympathetic nerve fibers in the epicardium of 4 normal controls, 11 cases with incidental Lewy bodies (iLBs), and 14 cases of PD. Cardiac sympathetic innervation was significantly less in PD than in normal controls and cases with iLBs (P < 0.05). There was also a decrease in TH-immunoreactive fibers in iLB cases compared to normal controls (P < 0.01). TH-immunoreactive fibers correlated with the PD stage (r = -0.75, P < 0.001), as well as with Hoehn & Yahr clinical stage (r = -0.61, P < 0.001), and disease duration (r = -0.63, P < 0.001). Immunohistochemistry for alpha-synuclein showed neurites in epicardium in PD and iLB cases, but not in normal controls. The density of alpha-synuclein neurites correlated with Braak PD stage (r = 0.38, P < 0.05), Hoehn & Yahr clinical stage (r = 0.44, P < 0.05), and disease duration (r = 0.42, P < 0.05). This study demonstrates that cardiac sympathetic degeneration and alpha-synuclein pathology is present in presymptomatic phase of PD, and that both increase with disease duration and severity.

The Persistent Sciatic Inflammatory Neuropathy (SIN) Rat Model of Neuropathic Pain Does Not Involve Small-Fiber Axon Damage

Most neuropathic pain conditions are caused by damage to peripheral nociceptive neurons known as “small-fibers.” Examination of punch skin biopsies immunolabeled with pan-axonal markers has identified degeneration of distal nociceptive axons as a pathological hallmark of most neuralgias including numerous polyneuropathies, postherpetic neuralgia, and complex regional pain syndrome. Similar axonopathy is present in several rodent models of neuralgia (chronic constriction injury and spared nerve injury). Here we have measured the density of distal cutaneous small-fiber axons in rats with persistent mechanical allodynia caused by the sciatic inflammatory neuropathy (SIN), a model of painful neuritis. We have evaluated whether perineural immune activation sufficient to cause evoked-pain behaviors is also associated with loss of distal epidermal innervation.SIN was created in adult male Sprague-Dawley rats by perineural zymosan administration using established methods. Unoperated rats provided controls. Two weeks of sensory testing established the presence of static mechanical allodynia. Full-thickness punches were then collected from ipsilesional sural-innervated plantar hindpaw skin. These were immunolabeled against PGP9.5 using standard methods, and the density of epidermal neurites quantitated microscopically by a single examiner who was unaware of rats' experimental group. Neurite densities were similar in SIN and control samples (P = 0.96), suggesting that mechanical allodynia can occur from inflammation near a nerve, without distal small-fiber degeneration.

Astrocytes, but not olfactory ensheathing cells or Schwann cells, promote myelination of CNS axons in vitro

We have examined the interaction between olfactory ensheathing cells (OECs), Schwann cells (SC), oligodendrocytes, and CNS axons using cultures generated from embryonic rat spinal cord. Oligodendrocyte process extension and myelination in these cultures was poor if the cells were plated on OECs or SCs. Myelin internodes and nodes of Ranvier formed frequently if these cultures were plated onto monolayers of neurosphere-derived astrocytes (NsAs). In the myelinated fibers generated on NsAs, Nav channels, caspr, and neurofascin molecules were correctly assembled at the nodes of Ranvier. The density of neurites, survival, and antigenic differentiation of oligodendrocytes was similar on OEC and NsAs monolayers. However, on OEC monolayers, despite a transient increase in the number of endogenous oligodendrocytes, there was a decrease in oligodendrocyte process extension and axonal ensheathment when compared with cultures plated on NsAs monolayers. To determine if these changes were due to axonal or glial factors, spinal cord oligodendrocytes were plated onto monolayers of OECs, NsAs, and poly-L-lysine in the absence of neurons. In these cultures, process extension and myelin-like membrane formation by oligodendrocytes was improved on monolayers of OEC. This suggests that inhibition of process extension is mediated via cross-talk between OECs and neurites. In cultures containing axons plated on OEC monolayers, oligodendrocyte process formation, axonal ensheathment, and myelination occurred albeit lower if the cultures were supplemented with NsAs conditioned medium. These data suggest OECs can permit neurite extension and oligodendrocyte proliferation, but lack secreted factor(s) and possible cell-cell contact that is necessary for oligodendrocyte process extension and myelination.

Brain tissue microarrays in neurodegenerative diseases: Validation of methodology and immunohistochemical study of growth‐associated protein‐43 and calretinin

A tissue microarray (TMA) was constructed using 47 neurodegenerative diseases (NDD), including Alzheimer's disease (AD; n = 30) and non-AD NDD (n = 17), and from seven controls. For validation of the methodology, the following three immunostains were used. Tau and beta-amyloid-related pathologies were more significantly recognized in tauopathies/AD compared to non-tauopathies/controls, and these results were comparable to the assessment of the whole brain sections. But no alpha-synuclein pathologies were observed despite five cases of dementia with Lewy bodies. It was concluded that the TMA technique is useful for NDD with diffuse pathological processes but not for those with patchy and occasional lesions or in early stages. Growth-associated protein (GAP)-43 and calretinin were also immunostained. A significant reduction in GAP-43 expression was seen in the frontal lobe and hippocampus in AD compared to non-AD cases, but not in other areas. No significant difference in number of anticalretinin immunoreactive neurons or in density of immunoreactive neurites was observed between any of the NDD and controls, which may indicate that calretinin-positive neurons are spared in the degenerative process. These results are compatible with the previous studies. These analyses were performed rapidly in a large number of cases using a single slide under uniform staining conditions.

FMRFamide-like immunocytochemistry in the brain and subesophageal ganglion of Triatoma infestans (Insecta: Heteroptera). Coexpression with β-pigment-dispersing hormone and small cardioactive peptide B

The distribution of FMRFamide (FMRFa)-like immunoreactivity (LI) was studied in the brain and subesophageal ganglion of Triatoma infestans, the insect vector of Chagas' disease. The neuropeptide displayed a widespread distribution with immunostained somata in the optic lobe, in the anterior, lateral, and posterior soma rinds of the protocerebrum, and around the antennal sensory and mechanosensory and motor neuropils of the deutocerebrum. FMRFa-immunoreactive profiles of the subesophageal ganglion were seen in the mandibular, maxillary, and labial neuromeres. Immunostained neurites were detected in the medulla and lobula of the optic lobe, the lateral protocerebral neuropil, the median bundle, the calyces and the stalk of the mushroom bodies, and the central body. In the deutocerebrum, the sensory glomeruli showed a higher density of immunoreactive processes than the mechanosensory and motor neuropil, whereas the neuropils of each neuromere of the subesophageal ganglion displayed a moderate density of immunoreactive neurites. Colocalization of FMRFa-LI and crustacean pigment-dispersing hormone-LI was found in perikarya of the proximal optic lobe, the lobula, the sensory deutocerebrum, and the labial neuromere of the subesophageal ganglion. The distribution pattern of small cardioactive peptide B (SCP(B))-LI was also widespread, with immunolabeled somata surrounding every neuropil region of the brain and subesophageal ganglion, except for the optic lobe. FMRFa- and SCP(B)-LIs showed extensive colocalization in the brain of this triatomine species. The presence of immunolabeled perikarya displaying either FMRFa- or SCP(B)-LI confirmed that each antisera identified different peptide molecules. The distribution of FMRFa immunostaining in T. infestans raises the possibility that FMRFa plays a role in the regulation of circadian rhythmicity. The finding of immunolabeling in neurosecretory somata of the protocerebrum suggests that this neuropeptide may also act as a neurohormone.

Estrogen alters amyloid precursor protein as well as dendritic and cholinergic markers in a mouse model of Down syndrome.

Individuals with Down syndrome (DS) develop the pathological hallmarks of Alzheimer's disease at an early age, later followed by memory decline and dementia. Women with DS are twice as likely to undergo early menopause, and levels of estradiol correlate with onset of cognitive decline in these women. We have demonstrated that a mouse model of DS, mice with segmental trisomy of chromosome 16 (Ts65Dn), develop a significant deficit in both reference and working memory as young adults (6-10 months of age), coupled with phenotypic loss of cholinergic neurons in the basal forebrain and altered growth factor levels. In the present study we examined cholinergic and dendritic markers in the hippocampal formation and levels of the amyloid precursor protein (APP) in different brain regions of Ts65Dn mice treated with estradiol for 60 days. The density of the dendritic marker Map2 was significantly decreased in the hippocampal formation of middle-aged trisomic mice (9-15 months old), and the density of cholinergic neurites (acetylcholinesterase [AChE] histochemistry) was also decreased in specific layers of the hippocampus. Treatment with 17beta-estradiol alleviated the decreases in Map2 and AChE staining, but had no effect on full-length APP levels in the hippocampus. In contrast, a main effect of treatment on APP levels in the striatum was noted, with significant elevations observed in controls and trisomics. These findings demonstrate that estrogen can alleviate deficits in cholinergic and dendritic elements in the hippocampal formation and further strengthens the rationale to explore estrogen replacement therapy in women with DS.

Medium requirements for neuritic outgrowth from goldfish retinal explants and the trophic effect of taurine

The use of culture media of known composition are necessary for studying the role of trophic molecules. Since most of the in vitro research on regeneration of the optic nerve has been performed in the presence of fetal calf serum, the aim of this study was to obtain a medium in which the neuritic outgrowth from post-crush goldfish retinal explants could take place without adding fetal calf serum. After the lesion of the optic nerve (10 days), the retina of goldfish was dissected and explants were cultured for 5 and 10 days in the absence or in the presence of fetal calf serum, at which time the neuritic outgrowth was determined. Various concentrations and combinations of glucose, albumin, calcium, HEPES and taurine were used. The highest neuritic outgrowth was observed in the presence of fetal calf serum, in which condition the amino acid taurine increased length and density of neurites. Media supplemented with albumin, calcium or HEPES did not modify the outgrowth of neurites from the explants. However, glucose favored the neuritic outgrowth in a bell-shaped manner, although fibers were thinner than those observed in the presence of fetal calf serum. Taurine did not stimulate outgrowth of neurites from explants growing in a medium with optimal concentrations of glucose, indicating that elements of the fetal calf serum are determinant for the trophic effect of taurine. The present results contribute to further studies, such as those related to the effect of taurine and of trophic factors derived from the optic tectum, which would be performed in the presence of a medium free of fetal calf serum.

Cortical Lewy body pathology in the diagnosis of dementia.

Dementia with Lewy bodies (DLB) and Parkinson's disease (PD) are distinguishable clinically but often not neuropathologically. This study aims to test whether the distribution of cortical Lewy bodies differs in these clinicopathological groups and to develop diagnostic protocols for their differentiation. Brains were obtained at autopsy from cases recruited from prospective clinical studies of dementia or movement disorders. All cases with significant pathologies other than Lewy bodies or plaques were excluded. Cases were categorised into either PD without dementia, DLB (dementia first or within 2 years of disease onset), or PD with a later onset of dementia (PDD). The distribution and density of Lewy bodies and Lewy neurites was determined using antibodies to ubiquitin and alpha-synuclein. Cortical Lewy body densities could not separate cases of DLB from those with PDD. However, semiquantitative thresholds in the parahippocampus could separate demented from non-demented cases with high sensitivity and specificity. Interactions between multiple pathologies were determined using factor analysis. Although many cases had CA2 Lewy neurites, this was not associated with severity or duration of either dementia or parkinsonism. Most DLB cases had significant plaque pathology, and severity and duration of dementia was related to both increasing parahippocampal Lewy body densities and neuritic plaque grade. Weighted kappa statistics revealed that the combination of these pathologies indicated a more severe dementia. These results suggest that dual pathologies cause DLB, and high densities of parahippocampal Lewy bodies indicate dementia regardless of additional pathologies.

Target-Dependent Sexual Differentiation of a Limbic–Hypothalamic Neural Pathway

Neural pathways between sexually dimorphic forebrain regions develop under the influence of sex steroid hormones during the perinatal period, but how these hormones specify precise sex-specific patterns of connectivity is unknown. A heterochronic coculture system was used to demonstrate that sex steroid hormones direct development of a sexually dimorphic limbic-hypothalamic neural pathway through a target-dependent mechanism. Explants of the principal nucleus of the bed nuclei of the stria terminalis (BSTp) extend neurites toward explants of the anteroventral periventricular nucleus (AVPV) derived from male but not female rats. Coculture of BSTp explants from male rats with AVPV explants derived from females treated in vivo with testosterone for 9 d resulted in a high density of neurites extending from the BSTp to the AVPV explant, as was the case when the BSTp explants were derived from females and the AVPV explants were derived from males or androgen-treated females. These in vitro findings suggest that during the postnatal period testosterone induces a target-derived, diffusible chemotropic activity that results in a sexually dimorphic pattern of connectivity.

The effects of taxol on the central nervous system response to physical injury.

Cytoskeletal disruption is a key pathological change in numerous human neurodegenerative diseases. We have, therefore, examined the effect of taxol, a microtubule-stabilising agent, on the neuronal response to localised trauma in the central nervous system utilising a rodent experimental model that replicates cytoskeletal alterations which occur in conditions such as Alzheimer's disease and head injury. At 1 day post-injury, 1 mM taxol administration to the damaged neocortex resulted in a statistically significant reduction in the density of abnormal neurites labelled with antibodies to neurofilaments. In addition, there was a relative preservation of MAP2 labelling of dendrites surrounding the injury site in taxol-treated, as compared to vehicle-treated, animals at 1 day post-injury. At 4 days post-injury, however, there was a statistically significant increase in the density of abnormal neurites surrounding the injury site in taxol-treated rats as compared to vehicle-treated animals. The degree of MAP2 labelling was also equally decreased in both vehicle- and taxol-treated animals as compared to normal cortex at this time point. Our data suggest that, in the short term, taxol may be stabilising neuronal microtubules and reducing reactive alterations in axons. After longer periods, however, our data indicate that the stereotypical neuronal reaction to trauma may be abnormally prolonged due to taxol administration, consistent with both in vivo work on taxol intoxication in the injured peripheral nervous system and in vitro culture studies.