An autopsy case of early (“minimal change”) olivopontocerebellar atrophy (multiple system atrophy-cerebellar)

We report a 57-year-old woman with multiple system atrophy (MSA) of 15-month duration. The patient developed dysarthria, followed by impaired balance of gait, mild limb ataxia, and saccadic eye movement. A postmortem examination performed after she was found dead in a bathtub revealed neuronal loss restricted to the olivopontocerebellar system, being more severe in the pontine nucleus. Mild neuronal loss was also found in the anterior vermis and inferior olivary nucleus. -Synuclein immunohistochemistry demonstrated widespread occurrence of glial cytoplasmic inclusions in the central nervous system, which were more numerous in the pontine base and cerebellar white matter. In contrast, neuronal -synuclein accumulation was confined to the pontine and inferior olivary nuclei. The number of neuronal intranuclear inclusions was much higher than that of neuronal cytoplasmic inclusions. Moreover, -synuclein accumulation was more severe in the neurites than in the cytoplasm or nucleus. This case demonstrates the early pattern of brain pathology in MSA-cerebellar (olivopontocerebellar atrophy).     

Extending the clinicopathological spectrum of neurofilament inclusion disease

We describe features of a patient that broadens the clinical and pathological spectrum of neurofilament inclusion disease (NFID). The patient was a 52-year-old man with a 5–6 year history of progressive, asymmetrical spastic weakness of the upper and lower extremities; l-DOPA-unresponsive parkinsonism; and SPECT evidence of asymmetrical frontoparietal and basal ganglia hypoperfusion. The brain had marked frontoparietal parasagittal cortical atrophy, including the motor cortex, with histopathological evidence of neurofilament- and -internexin-immunoreactive neuronal inclusions. The corticospinal tract had degeneration, but there was minimal lower motor neuron pathology. There was also severe neuronal loss and gliosis in the posterolateral putamen and the substantia nigra, mimicking multiple system atrophy; however, glial cytoplasmic inclusions were not detected with -synuclein immunohistochemistry. This case extends the clinical and pathological spectrum of NFID to include cases with predominant parkinsonian and pyramidal features.Gerry Shaw holds equity in EnCor Biotechnology Inc., a company commercializing the alpha-internexin antibody used in this study, and may benefit by receiving royalties or equity growth.     

Peripheral noradrenergic function during chronic lithium treatment in the rat

Summary Noradrenergic function was evaluated in the rat vas deferens following chronic treatment with lithium (30 mM in the diet for 2 weeks followed by 50 mM for 1 week). No alteration could be detected in: a)³ H-noradrenaline release evoked by electrical field stimulation, b) ₁adrenoceptor function as assessed by contractile response to noradrenaline, c) active neuronal uptake of³ H-noradrenaline, d) ²-adrenoceptor activity, as assessed by yohimbine-enhancement of evoked release of³ H-noradrenaline. The data argue against a primary action of lithium on the noradrenergic nerve. On the other hand, these findings do not rule out the possibility that the function of CNS noradrenergic neurons could be altered as a result of the known effects of lithium on the phosphatidyl-inositol system, and the greater sensitivity of CNS neurons to depletion of the neuronal membrane inositol pool.     

Accumulation of phosphorylated α-synuclein in the brain and peripheral ganglia of patients with multiple system atrophy

We immunohistochemically examined the brain and peripheral sympathetic ganglia from eight patients with multiple system atrophy (MSA), using an antibody specific for phosphorylated -synuclein (anti-PSer129). Phosphorylated -synuclein was deposited in five cellular locations: oligodendroglial cytoplasm and nucleus, and neuronal cytoplasm, processes and nucleus. Many neuronal cytoplasmic inclusions (NCIs) were found in the pontine and inferior olivary nuclei and, to a lesser extent, in the substantia nigra, locus ceruleus, and neocortical and hippocampal neurons. NCIs were also found in the sympathetic ganglia in two out of the eight cases. Moreover, anti-PSer129 immunohistochemistry revealed extensive neuropil pathology; swollen neurites were abundant in the pontine nucleus, delicate neurites were observed in the deeper layers of the cerebral cortex and thalamus, and neuropil threads and dot-like structures were distributed in the basal ganglia and brainstem. Diffuse neuronal cytoplasmic staining (pre-NCI) was frequently found in the pontine and inferior olivary nuclei. Thus, the widespread accumulation of phosphorylated -synuclein in both glial and neuronal cells is a pathological feature in patients suffering from MSA.     

Neuropathological findings of an autopsy case of adult β-galactosidase and neuraminidase deficiency

Summary An autopsy case of a Japanese male with familial -galactosidase and neuraminidase deficiency is reported. The clinical picture was characterized by adult onset, a gargoyle-like face, cerebellar ataxia, myoclonus, convulsions, retinal degeneration and cortical blindness.Histopathologically, most neurons seemed to have become degenerated in the whole cerebral cortex. Moreover, the calcarine cortex appeared spongy with depopulation of nerve cells. Stuffed neurons or neuronal storage changes were found throughout the brain, especially in the motor nuclei of the spinal cord and brain stem.The inclusions in the stuffed neurons revealed various profiles on the electron microscope. They were composed of membranous lamellar and/or multilamellar structures, often accompanying vacuoles and reminiscent of lipofuscin-like profiles.     

Characterisation of a functional polyamine site on rat mast cells: association with a NMDA receptor macrocomplex

Polyamines can modulate activation of N-methyl-d-aspartate (NMDA) receptors by binding to a specific polyamine site associated with a NMDA receptor macrocomplex. Polyamines induce histamine release from mast cells, although the mechanism had not been defined. We have examined whether spermine, a natural polyamine, and compound , regarded as a synthetic polyamine, activate mast cells by a polyamine site associated with a NMDA receptor macrocomplex. Spermine induced secretion of histamine from rat peritoneal mast cells and rat brain mast cells in a concentration-dependent manner. Rat peritoneal mast cells were used as a model system to explore the effects of NMDA antagonists on polyamine-induced histamine release. Ifenprodil, MK801 and arcaine inhibited histamine secretion from mast cells exposed to polyamines; the percentage inhibition was greater against spermine than compound . These data support the proposal that spermine (and possibly compound ) induce histamine release from mast cells by interacting with a specific polyamine site on a NMDA receptor complex.     

Neuronal intranuclear inclusions are ultrastructurally and immunologically distinct from cytoplasmic inclusions of neuronal intermediate filament inclusion disease

Abnormal neuronal cytoplasmic inclusions (NCIs) containing aggregates of -internexin and the neurofilament (NF) subunits, NF-H, NF-M, and NF-L, are the signature lesions of neuronal intermediate filament (IF) inclusion disease (NIFID). The disease has a clinically heterogeneous phenotype, including frontotemporal dementia, pyramidal and extrapyramidal signs presenting at a young age. NCIs are variably ubiquitinated and about half of cases also have neuronal intranuclear inclusions (NIIs), which are also ubiquitinated. NIIs have been described in polyglutamine-repeat expansion diseases, where they are strongly ubiquitin immunoreactive. The fine structure of NIIs of NIFID has not previously been described. Therefore, to determine the ultrastructure of NIIs, immunoelectron microscopy was undertaken on NIFID cases and normal aged control brains. Our results indicate that the NIIs of NIFID are strongly ubiquitin immunoreactive. However, unlike NCIs which contain ubiquitin, -internexin and NF epitopes, NIIs contain neither epitopes of -internexin nor NF subunits. Neither NIIs nor NCIs were recognised by antibodies to expanded polyglutamine repeats. The NII of NIFID lacks a limiting membrane and contains straight filaments of 20 nm mean width (range 11–35 nm), while NCIs contain filaments with a mean width of 10 nm (range 5–18 nm; t-test, P<0.001). Biochemistry revealed no differences in neuronal IF protein mobilities between NIFID and normal brain tissue. Therefore, NIIs of NIFID contain filaments morphologically and immunologically distinct from those of NCIs, and both types of inclusion lack expanded polyglutamine tracts of the triplet-repeat expansion diseases. These observations indicate that abnormal protein aggregation follows separate pathways in different neuronal compartments of NIFID.     

Electron microscopic studies on the cerebral lesions of rats in experimental chronic disulfiram poisoning

Summary Following chronic administration of disulfiram to rats, changes of the brain were examined electron-microscopically. Pathological findings were observed in the nerve cells of the cerebral cortex and hypothalamus at later stage, and synaptic changes in the hypothalamus from initial stage. On the other hand, changes of myelinated fibers, neuroglias and capillaries were very slight. It was considered that neurons were affected more predominantly than other neuronal elements by the cytotoxic action of the drug, and that the synaptic changes of the hypothalamus might reveal chronic disturbance of noradrenergic transmission by inhibition of dopamine--hydroxylase. These ultrastructural findings might relate to the pathogenic mechanism of the disulfiram psychosis.     

Genetic and environmental factors in the pathogenesis of Huntington’s disease

Huntingtons disease is a fatal inherited disorder in which there is progressive neurodegeneration in specific brain areas, mainly the striatum and cerebral cortex, producing motor, cognitive, and psychiatric symptoms. The trinucleotide repeat mutation involved is common to many other brain diseases, which may therefore involve similar mechanisms of pathogenesis. We are beginning to understand how a CAG trinucleotide repeat expansion in the disease gene, encoding an expanded polyglutamine tract, induces neuronal dysfunction and symptomatology in Huntingtons disease. Recent evidence that environmental factors modify the onset and progression of neurodegeneration has shed new light on Huntingtons disease and other devastating brain diseases. This review focuses on genetic mediators, environmental modulators, and associated gene-environment interactions in the pathogenesis of Huntingtons disease.     

Monoclonal antibody to β peptide,recognizing amyloid deposits,neuronal cells and lipofuscin pigments in systemic organs

Summary A monoclonal antibody (AmT-1) produced against synthetic amyloid peptide (1–28 residues) was revealed to be reactive with amyloid peptide blotted on nitrocellulose membrane, but not with that dissolved in sodium dodecyl sulfate and electrophoresed. AmT-1 immunostained senile plaques of typical, primitive and diffuse type, as well as amyloid deposits in cerebral vessels. It also reacted with neuronal and glial cells of normal and Alzheimer's disease (AD) brains. In addition, AmT-1 was also reactive strongly with lipofuscin pigments of adrenal reticular cells, and weakly with those of eccrine glands and liver cells. A rat neural cell line (PC12h) was reactive with AmT-1. By immunoelectron microscopy, a positive reaction was seen in ribosomes along the rough endoplasmic reticulum of nerve cells and PC12h cells. By immunoprecipitation, AmT1 reacted with a band at 36 kDa in the brain homogenates from Ad patients as well as from normal aged subjects. By immunoblotting analysis, AmT1 reacted with a band at 36 kDa in the cytosolic fraction of PC12 cells, and three bands (12–17 kDa) in the lipopigment fraction of the adrenal gland. These findings suggest that the cerebral amyloid deposits contain substance(s) having an epitope common to neuronal cells and lipofuscin pigments. The possible relationship between cerebral amyloid deposits and lipofuscin pigments in systemic organs is discussed.     

Familial frontotemporal dementia: a report of three cases of severe cerebral atrophy with rare inclusions that are negative for tau and synuclein,but positive for ubiquitin

We describe the brain autopsy findings from three of five siblings who suffered dementia with clinical diagnoses including Alzheimers, Parkinsons, and Picks disease. Five other living siblings appear unaffected. All of the autopsied brains showed severe atrophy (brain weights 613, 641, and 750 g) of the frontal and temporal lobes, and to a slightly lesser extent of the parietal lobes, while the occipital lobes were relatively preserved. The substantia nigra showed marked neuronal loss with gliosis. No ballooned neurons, neurofibrillary tangles, neuritic plaques, Pick bodies, or Lewy bodies were found in these brains. Immunohistochemistry for tau protein failed to reveal neuronal or glial inclusions, and normal tau protein was found in a separate Western blot study [Adamec et al. (2001) Neurosci Lett 315:21–24]. Rare neurons with ubiquitinated cytoplasmic inclusions were scattered in the neocortex and hippocampus. The overall pathological features were consistent with a severe form of frontotemporal dementia (FTD) with involvement of the substantia nigra. Whether the rare ubiquitinated inclusions are sufficient to classify these cases as FTD with motor neuron disease type inclusions but without motor neuron disease, or FTD dementia lacking distinctive histological features remains unclear. The features of lobar circumscribed atrophy without Pick bodies and without ballooned neurons, however, are consistent with Pick disease group C in the Constantinidis classification [Constantinidis et al. (1974) Eur Neurol 11:208–217].     

Neuroprotective effects of α-dihydroergocryptine against damages in the substantia nigra caused by severe treatment with 1-methyl-4-phenyl-1,3,3,6-tetrahydropyridine

Summary The effects on the substantia nigra of -dihydroergocryptine (DEK), a drug with strong dopaminomimetic activity, were tested with a severe 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment in monkeys. Compared with monkeys treated with MPTP alone, the animals which received DEK plus MPTP showed reduced neuronal death in the substantia nigra. The reactive astrocytes were increased in number. Moreover, several axons which were immunopositive to phosphorylated neurofilament proteins and with features similar to those of control animals were seen in the pars compacta. The findings suggest that DEK preserves neuronal morphology and brain architecture.supported by Italian M.U.R.S.T. grants (to G.B.) and by Poli S. P. A., Rozzano, Milano     

Alpha-synuclein pathology in Parkinson's and Alzheimer's disease brain: incidence and topographic distribution--a pilot study

To study the incidence and topographic distribution of -synuclein-positive inclusions in Parkinsons disease (PD), dementia with LB (DLB), and Alzheimers disease (AD), 206 brains of elderly patients, including 53 patients with clinical PD, 110 autopsy-proven AD cases, 22 with dementia with LB (DLB), 1 case with essential tremor, and 20 age-matched controls were investigated using -synuclein immunohistochemistry. For technical reasons, the olfactory system was not studied. In all PD brains, -synuclein-positive inclusions and neuronal losses were present in medullary and pontine nuclei, locus coeruleus, and substantia nigra, with additional lesions in amygdala (24%), allocortex (58%), cingulate area (34%), and isocortex (26.5%). All PD cases corresponded to pathology stage 4–6 suggested by Braak et al. (2003, Neurobiol Aging 24:197). In most cases of DLB, the distribution of -synuclein pathology and neurodegeneration corresponded to stages 5 and 6 of PD pathology. The case with essential tremor and 48.2% of the AD cases showed no LB pathology; in the other AD brains -synuclein-positive inclusions were seen in various brain areas. None of the controls showed LB pathology. Among 12 cases of incidental Lewy body disease (without clinical parkinsonian signs), 7 corresponded morphologically to PD stage 3 or 4. In further 6 AD cases, 2 with parkinsonian symptoms, considerable damage to locus coeruleus, substantia nigra, nucleus basalis and allocortex with preservation of the medullary nuclei was seen. The preliminary data largely confirm the Braak staging of brain pathology, although some of the clinical PD cases corresponded to stage 3 often considered as preclinical. In addition, some cases without demonstrable involvement of medullary nuclei showed extensive PD-like pathology in other brain areas, suggesting deviation from the proposed stereotypic expansion pattern and that incidental LB pathology may affect solely the locus coeruleus and substantia nigra. Striking similarity of LB pathology between DLB and PD suggests close morphological relationship between both disorders. Widespread LB lesions occurred in many sporadic AD cases without parkinsonian symptoms, the pathogenesis and clinical impact of which are unclear. The relationship between AD and PD with particular reference to synaptophysin-positive lesions needs further elucidation.An erratum to this article can be found at     

Energy gradients for the homeostatic control of brain ECF composition and for VT signal migration: introduction of the tide hypothesis

Summary. The present paper enlightens a new point of view on brain homeostasis and communication, namely how the brain takes advantage of different chemical-physical phenomena such as pressure waves, and temperature and concentration gradients to allow the renewal of the extra-cellular fluid (i.e., the homeostasis of the brain internal milieu) as well as some forms of intercellular communications (Volume Transmission) at an energy cost much lower than the classical synaptic transmission (the prototype of Wiring Transmission).In particular, the possible functional meaning of the intracranial pressure waves is discussed in the frame of the so called tide hypothesis which maintains that the pressure waves, created by the cardiac pump, modulate the cerebro-spinal fluid flow from and towards the subarachnoid space as well as towards and from the Virchow-Robin spaces. These fluid push-pull movements favor both the migration of signals and the extra-cellular fluid renewal, especially in the cerebral cortex.Dedicated to Prof. B. Magnani, Professor of Cardiology, Medical Faculty of Bologna, Italy     

α-Internexin aggregates are abundant in neuronal intermediate filament inclusion disease (NIFID) but rare in other neurodegenerative diseases

Abnormal neuronal aggregates of -internexin and the three neurofilament (NF) subunits, NF-L, NF-M, and NF-H have recently been identified as the pathological hallmarks of neuronal intermediate filament (IF) inclusion disease (NIFID), a novel neurological disease of early onset with a variable clinical phenotype including frontotemporal dementia, pyramidal and extrapyramidal signs. -Internexin, a class IV IF protein, a major component of inclusions in NIFID, has not previously been identified as a component of the pathological protein aggregates of any other neurodegenerative disease. Therefore, to determine the specificity of this protein, -internexin immunohistochemistry was undertaken on cases of NIFID, non-tau frontotemporal dementias, motor neuron disease, -synucleinopathies, tauopathies, and normal aged control brains. Our results indicate that class IV IF proteins are present within the pleomorphic inclusions of all cases of NIFID. Small subsets of abnormal neuronal inclusions in Alzheimers disease, Lewy body diseases, and motor neuron disease also contain epitopes of -internexin. Thus, -internexin is a major component of the neuronal inclusions in NIFID and a relatively minor component of inclusions in other neurodegenerative diseases. The discovery of -internexin in neuronal cytoplasmic inclusions implicates novel mechanisms of pathogenesis in NIFID and other neurological diseases with pathological filamentous neuronal inclusions.     

Immunohistological study of senile brains by using a monoclonal antibody recognizing β amyloid precursor protein: significance of granular deposits in relation with senile plaques

Summary Immunochemical analyses revealed that a monclonal antibody Am-3 recognized amyloid precursor protein (APP) in senile plaques extracted from Alzheimer's brain, but did not recognize amyloid protein. Immunohistochemically, however, the staining pattern of Am-3 in frozen section of Alzheimer's brain was almost the same with that of rabbit polyclonal antibody to amyloid peptide which could recognize both amyloid protein and APP. In other words, APP was present in senile plaques of various types, cerebrovascular amyloid and granular deposits. The granular deposits were 5–10 m in size and laminarily distributed in the 1st, 3rd and 4th layers of cerebral cortex. They were especially abundant in 1st and 4th layers where senile plaques were usually fewer in number. Although the distribution in the cerebral cortex was different between the senile plaques and the granular deposits, the number of the granular deposits was well correlated with that of senile plaques. The granular deposits were negative in Congo-red birefringence, but contained amyloid protein as well as APP fragment judging from positive staining by both Am-3 and polyclonal antibody to synthetic amyloid peptide. Thus, they could be regarded as pre-amyloid.     

δ-Aminolaevulinic acid dehydratase activity and focal brain haemorrhages in lead-treated rats

Summary Mothers were fed a diet containing 2% lead acetate according to the Pentschew-Garro model for inducing lead encephalopathy in young rats. At 20–22 days of age the young lead-treated rats had a mean brain Pb of 2.8 g/g and liver Pb of 11 g/g. The ALA dehydratase activity decreased 29% in brain and 69% in liver compared to controls, suggesting that the enzyme activity is related to the tissue lead level.Mothers that had received lead prior to conception gave birth to pups with a significantly raised mean blood lead level (44 g%). The ALA dehydratase activity in brain and liver was unchanged, suggesting that low blood lead levels may be insufficient to inhibit this enzyme in the rat. Focal haemorrhages were present, however, in the cerebral cortex of some of the pups from the lead-treated mothers. It is concluded that damage to the rat brain vascular system is a better index of lead toxicity than measurement of the lead sensitive enzyme ALA dehydratase.     

α-Synuclein in motor neuron disease: an immunohistologic study

-Synuclein (ASN) has been implicated in neurodegenerative disorders characterized by Lewy body inclusions such as Parkinsons disease and dementia with Lewy bodies. Lewy body-like inclusions have also been observed in spinal neurons of patients with amyotrophic lateral sclerosis (ALS) and reports suggest possible ASN abnormalities in ALS patients. We assessed ASN immunoreactivity in spinal and brain tissues of subjects who had died of progressive motor neuron disorders (MND). Clinical records of subjects with MND and a comparison group were reviewed to determine the diagnosis according to El-Escariol Criteria of ALS. Cervical, thoracic and lumbar cord sections were stained with an antibody to ASN. A blinded, semiquantitative review of sections from both groups included examination for evidence of spheroids, neuronal staining, cytoplasmic inclusions, anterior horn granules, white and gray matter glial staining, corticospinal tract axonal fiber and myelin changes. MND cases, including ALS and progressive muscular atrophy, displayed significantly increased ASN staining of spheroids (P0.001), and glial staining in gray and white matter (P0.05). Significant abnormal staining of corticospinal axon tract fibers and myelin was also observed (P0.05 and 0.01). Detection of possible ASN-positive neuronal inclusions did not differ between groups. Significant ASN abnormalities were observed in MND. These findings suggest a possible role for ASN in MND; however, the precise nature of this association is unclear.     

Pharmacokinetics and pharmacodynamics of -arginine in rats: a model of stimulated neuronal nitric oxide synthesis

Nitric oxide (NO) is believed to be involved in a variety of central nervous system (CNS) functions, including opioid responsivity. Elucidation of the role of NO in the CNS requires the ability to elevate systematically neuronal NO concentrations in vivo. This study was conducted to assess the pharmacokinetics of -arginine, a NO precursor, and to relate the disposition of this amino acid to the pharmacodynamic endpoint of neuronal NO production. -Arginine (250-, 500-, or 1000-mg/kg/h) or saline was infused intravenously for 6 h to rats. -Arginine was quantified in brain and blood (after in vivo microdialysis) with high-performance liquid chromatography. NO was quantified simultaneously with a sensitive and specific amperometric sensor placed in the hippocampus. The data were fit with a comprehensive pharmacokinetic–pharmacodynamic (PK/PD) model to obtain parameters governing the systemic disposition of -arginine, the uptake of -arginine into the brain, and subsequent NO production. Exogenous administration of -arginine resulted in incremental elevations in hippocampal NO, with a 33, 48, and 50% increase from control for the 250-, 500-, and 1000-mg/kg/h -arginine treated rats, respectively. The PK/PD model, which incorporated known characteristics of the system (saturable uptake of -arginine into brain; NO production governed by circadian changes in enzyme activity) was capable of describing accurately the observed data. The model developed herein will be invaluable in characterizing the numerous roles of NO in the CNS.     

Studies on GM1-gangliosidosis,type II

Summary Post-mortem studies on a 6-year old boy with G_M1-gangliosidosis, Type II revealed no evidence of accumulation of residual bodies nor of gangliosides or glycoproteins in liver and spleen. In brain tissue the ganglioside G_M1 accounted for 70% of the ganglioside fraction and ganglioside-NANA was increased 3.6 fold over controls. In addition, the brain tissue contained large amounts of glycoprotein, glycoprotein derived galactose being increased 2.5 times. The neuronal accumulation of tertiary lysosomes exhibited a characteristic distributional pattern: in general the large neuronal perikarya were more consistently involved with the exception of the motor cells of the cranial nerve nuclei, III, IV, and VI. In addition to characteristic MCB's, the nerve cells contained residual bodies with a granulo-floccular matrix, presumed to represent glycoproteins.The distribution of the mutant gene was studied among 30 blood relatives of the proband at risk and 6 carriers could be ascertained on the basis of a reduced leukocytic -galactosidase activity.The partly purified enzyme from the patient's liver revealed 20% activity as compared to that of normal controls. All three fractions obtained by DEAE cellulose column chromatography exhibited markedly reduced activity at pH 3.6, but nearly normal activity at pH 6.6. The reduced activity corresponded to the B component of the enzyme as shown by electrophoretic separation.It is pointed out that this case cannot be diagnosed as generalized gangliosidosis for the process of ganglioside accumulation was restricted to nervous tissue.This work was supported by USPHS Grants NS-04607 and NS-08639 and by a grant from Children's Brain Diseases, San Francisco, CA.