Morphological abnormalities of erythrocyte membrane in the hereditary neurological disease with chorea, areflexia and acanthocytosis

A study with freeze-fracture electron microscopy was made on erythrocyte membrane in 5 patients with the hereditary neurological disease with chorea, areflexia, and acanthocytosis (Levine et al. 1968). Fixed or unfixed specimens from circulating blood were processed by the freeze-fracture technique, and the distribution of the intramembranous particles was studied. A significant increase in areas without intramembraneous particles (IMP-free areas) was found in all cases with the disease as compared to normal subjects. The group mean value of IMP-free areas was 0.11 +/- 0.01 X 10-2 micrometer2/micrometer2 fracture face in the normal P face and 0.10 +/- 0.04 in the normal E face, compared with 1.77 +/- 0.49 in the P face and 1.33 +/- 0.17 in the E face in material from patients. Although abnormalities in lipid metabolism or membrane properties of erythrocytes have not been revealed in this condition hitherto, the present data suggest an abnormality of membrane fluidity due to abnormal lipid metabolism in this unique disease with neurological disorders and acanthocytosis.     

Immunoglobulins in cerebrospinal fluid: Enzyme-immunoassay

Using an enzyme-immunoassay (EIA) technique we established control values for IgA and IgM in cerebrospinal fluid (CSF). These values, together with IgG values determined by single radial immunodiffusion (SRID) technique, showed significant positive correlations with CSF total protein values. The CSF IgA and IgM levels were related to corresponding levels determined in serum. In addition, the values for all 3 immunoglobulin classes in CSF as well as CSF total protein values showed positive correlation with age of subjects, and IgG% and IgA% increased with age. This new EIA procedure can be completed within 24 h and is sensitive enough for determining all 3 immunoglobulin classes using a small amount (100 μl or less) of native CSF.     

Ultrastructural changes in muscle and motor end-plate of the dystrophic mouse

A comparative study of dystrophic mice (C57BL/6J-dy2J) and normal littermates, 6 to 9 months old, has revealed numerous ultrastructural changes in the dystrophic soleus. Vacuoles, swollen mitochondria, vesicular aggregates, membranous bodies, Z-line degradation, localized hypercontraction, myofibrillar disorientation, and focal necrosis were common. We report here new observations. Discontinuous, dense bands (240 A wide) were observed rarely between inner and outer membranes in some nuclei. Closely associated with infoldings of nuclear membrane were paracrystalline, vesicular structures, actin-like filaments, and autophagic vacuoles. With the degeneration of the nuclear membrane, actin-like filaments were observed within the nucleus. The primary synaptic clefts were widened and contained osmophilic-dense granules (diameter 0.2 to 0.7 micrometer). There was loss of secondary synaptic folds. Discontinuity of pre- and postsynaptic membranes could be observed, though rarely. Lysosome-like dense bodies were present among myofilaments. The myeloid bodies and autophagic vacuoles that were associated with nuclei, sarcoplasmic reticulum, and mitochondria gave a positive acid phosphatase reaction. Such degeneration changes were not observed in the normal soleus.     

Increased calcium-activated neutral protease activity in muscles of dystrophic hamsters and mice

A Ca²⁺-activated neutral protease activity was examined in muscles of normal and dystrophic hamsters and mice. Light grey and golden brown strains of normal and B10 14.6 strain of dystrophic hamsters were used. Normal and dystrophic mice were of the Bar Harbor 129 ReJ strain. Enzyme activity was measured in the post myofibrillar fraction (homogenate) and in the 75,000 × g pellet (particulate fraction) and supernatant using purified myofibrils.

In normal and dystrophic hamsters or mice, the Ca²⁺-activated neutral protease was most active in the supernatant followed by the homogenate and particulate fractions. As compared to fractions from normal muscle, enzyme activity was significantly elevated in all 3 fractions from dystrophic muscles of hamsters and mice. Both homogenate and supernatant fractions from muscles of normal hamsters had significantly higher enzyme activity than those of normal mice. Enzyme activity was similar in the particulate fraction. Similarly enzyme activity in the 3 fractions from dystrophic hamster and mouse muscles showed no significant difference.

It is suggested that the Ca²⁺-activated neutral protease may be involved in muscle fibre necrosis in muscular dystrophy.     

"Myopathic" changes in chorea-acanthocytosis. Clinical and histopathological studies

Four cases of chorea-acanthocytosis were studied with special reference to muscular changes. All the cases showed the clinical stigmata of oro-linguo-facial dyskinesia with tongue biting, mild neurogenic muscular involvement and acanthocytosis. Serum creatine kinase (CK) was persistently elevated, showing MM type isozyme predominance. Histopathological studies of the peroneus brevis muscle showed prominent small group atrophy, increase of small fibers on diameter analysis, frequent angulated fibers, and angulated fibers with increased acid phosphatase activity. These findings are compatible with chronic denervation. However, central nucleation (approximately 10%) and fiber splitting (2-8%) were also found in all cases. These are compatible with myopathic changes. No correlation of these "myopathic" changes and serum CK levels was found. The "myopathic" findings are probably secondary to chronic denervation.     

Human monoclonal antibodies to myelin-associated glycoprotein : Comparison of specificity and use for immunocytochemical localisation of the antigen

Recent reports demonstrate that a population of patients with gammopathy and demyelinating neuropathy have monoclonal antibodies to myelin-associated glycoprotein. Using the immunoblotting technique we compared the species specificity: human monoclonal antibodies to myelin-associated glycoprotein reacted with human, monkey, calf, dog, rabbit and guinea pig myelin, but not with rat or mouse. On the immunoblot myelin-associated glycoprotein consistently stained as a diffuse band with an apparent molecular weight ranging from 90-100 X 10(3) dalton. Experiments showed that when human CNS myelin had been incubated at 37 degrees C before gel electrophoresis, there was a general shift of staining towards the lower end of the molecular weight range. This low molecular weight myelin-associated glycoprotein, when released from the membrane, contains the antigenic determinant. Peptide mapping by limited proteolysis reveals that the antigenic determinants for 4 different monoclonal antibodies appear to lie very close together in the molecule. The distribution of the antigen was studied in nervous tissue with the unlabelled peroxidase-antiperoxidase method. The results obtained are in close agreement with the localization reported with polyclonal antisera to myelin-associated glycoprotein.     

Stimulation of the cerebral peduncles modulates tooth pulp-evoked firing of trigeminal caudalis neurons

Unit activity was recorded from trigeminal subnucleus caudalis neurons in immobilized, lightly anesthetized (Nembutal) cats following bipolar electrical stimulation of the canine tooth pulp. Electrical stimuli applied to the cerebral peduncles through stereotaxically placed coaxial electrodes discharged or inhibited most tooth pulp-activated nucleus caudalis (relay and nonrelay) neurons.     

Immune response to isolated oligodendrocytes

Oligodendrocytes were isolated from bovine white matter and were injected with complete Freund's adjuvant (CFA) into experimental animals. Indirect immunofluorescence studies using fluoresceinated goat anti-rabbit or anti-guinea pig immunoglobulin (GARIg; GAGPIg) showed that rabbit and guinea pig anti-oligodendrocyte (RAO, GPAO) sera reacted specifically with the surface of isolated oligodendrocytes in suspension, as well as with oligodendroglia in bovine and human brain sections, and in mouse cerebellum cultures. This activity of RAO was blocked by non-fluoresceinated GARIg and by GPAO, and absorbed by oligodendrocyte preparation (OP) or whole white matter, but not by purified myelin, neuroblastoma or non-brain tissue. Low levels of anti-basic protein antibodies were found in many RAO (but not GPAO) sera by radioimmunoassay, and a few showed significant anti-galactocerebroside antibody by agglutination and radioimmunoprecipation techniques. Guinea pigs sensitized with isolated oligodendrocytes in CFA showed cell-mediated immunity (CMI) to OP as manifested by delayed type skin test and induced in vitro lymphocyte transformation. CMI to purified myelin basic protein was not detected. The demonstration of humoral and CMI to the cell responsible for the production of CNS myelin may be related to some aspects of the immunopathogenesis of demyelinating disorders.     

Distribution of arsenic,manganese, and selenium in the human brain in chronic renal insufficiency,Parkinson's disease,and amyotrophic lateral sclerosis

The concentrations of arsenic, manganese and selenium/g wet tissue weight were determined in samples from 24 areas of the human brain from 3 patients with chronic renal insufficiency, 2 with Parkinson's disease and 1 with amyotrophic lateral sclerosis. The concentrations of the 3 elements were determined for each sample by neutron activation analysis with radiochemical separation. Overall arsenic concentrations were about 2.5 times higher in patients with chronic renal failure than in controls, and lower than normal in the patients with Parkinson's disease and amyotrophic lateral sclerosis. There were no obvious differences in the overall concentrations of manganese and selenium from one group to another. Even multivariate data analysis by the SIMCA method failed to reveal any significant difference in the distribution pattern of manganese and selenium in Parkinson's disease compared to normal controls.     

Ouabain binding sites in skeletal muscle from normal and dystrophic mice.

The specific binding of tritiated ouabain was used to estimate the density of Na+-K+-ATPase sites ("Na+-pump" sites) in segments of skeletal muscle from normal and dystrophic mice. Ouabain binding was approximately 4 times greater in red (soleus) muscle than in white (superficial gastrocnemius) muscle from normal animals. In dystrophic soleus muscles, ouabain binding was decreased by nearly one-half. Because Na+-K+-ATPase activity is associated with plasma membranes, these observations constitute further evidence for a sarcolemmal abnormality in dystrophic mice.     

Morphometric quantification of the cervical limb motor cells in controls and in amyotrophic lateral sclerosis.

The limb motor cells of the C6 segment of the spinal cord were counted and correlated with quantified histological findings of biceps brachii muscles in controls and in cases of amyotrophic lateral sclerosis (ALS). In 12 controls the motor cells were divided into larger ones with a minimum diameter greater than 20 micron, and smaller cells. Total numbers of the larger motor cells decreased significantly in 11 of 12 cases of ALS and of the smaller cells in 4 cases. In 4 controls most of the constituents of the biceps brachii muscle were normal-sized fibers, while in ALS smaller fibers and interstitial connective tissue increased and hypertrophic fibers decreased in association with a decrease of normal-sized fibers. The correlation coefficients between total numbers of the larger or smaller motor cells and normal-sized fibers in ALS were 0.92 and 0.65 respectively, and the larger motor cells, correlating with muscular atrophy of the upper arm, were considered to be alpha motor cells. Although in ALS the larger motor cells decreased almost diffusely, there were segmental variations, similar to controls, in numbers of the motor cells per 500 micron thickness.     

Relationship between physical findings and related subjective symptoms in neuropsychiatric patients with somatic complications—With special reference to ophthalmologic observations

The relationship between physical signs and subjective symptoms was examined on ophthalmological abnormalities in 306 neuropsychiatric inpatients, mainly in schizophrenics. The average duration of disease was 11.3 years and chronic cases were prominent. Control cases were 89 patients with pulmonary tuberculosis. Spontaneous complaints of the eye were made in four of the 306 cases. At ophthalmological examination a variety of abnormal findings were revealed in 268 cases (87.6%). This fact proved that neuropsychiatric patients complained rarely as compared with the rate of abnormal findings. As a result of clinical interview using a definite questionnaire, subjective symptoms were found in 154 (50.3%), being significantly lower than 69 (77.5%) in tuberculosis patients (p < 0.05). Accordance between subjective symptoms and abnormal findings was found more often in cases of tuberculosis than in cases of neuropsychiatric patients (p < 0.005). Among cases showing discordance, absence of subjective symptoms in spite of presence of abnormal findings, was observed more frequently in neuropsychiatric patients (p < 0.005), especially in schizophrenics with highly advanced emotional disorder. Abnormal findings which were apt to be accompanied with no subjective symptoms were disorders of visual acuity, and opacities of the cornea and the lens. Existence of subjective symptoms without any abnormal findings was more remarkable in the tuberculosis patients (p < 0.005) than in the neuropsychiatric patients.In the study of the relation between absence of subjective symptoms and administered psychotropic drugs, the amount of drugs administered during the day of ophthalmological examination, the period from the beginning of therapy to the examination (whole administration period), the total amount of psychotropic drugs and others were calculated. Cases having no subjective symptoms in spite of presence of abnormal findings presented higher levels of drug dosage and also longer periods in the whole term than cases having subjective symptoms corresponding to abnormal findings, but there was no significant difference.Comments were made on the results decribed above, emphasizing the importance of objective physical examinations during the treatment of neuropsychiatric disorders.     

Abnormalities of the sciatic nerves of dystrophic mice with reference to the large U-axons

A statistical study using regression analysis was used to evaluate the density of axonal organelles in dystrophic peripheral nerves. The slope of the density of neurotubules (NT) in myelinated (M-) axons was different from that in small unmyelinated (U-) axons. The slope of the density of NT in large U-axons (larger than 1.5 micron in diameter) was similar to that of the M-axons in both the dystrophic and control mice. There was a higher density of NT in the dystrophic M-axons than in the controls in the anterior, posterior and mixed nerves of the sciatic nerve. There was also a higher density of NT in the dystrophic small U-axons than in the controls. There was a higher density of neurofilaments (NF) of M-axons in the dystrophic mice than in the controls. On the contrary, the NF of small U-axons were lower in density in the dystrophic mice. These results were different from our previous reports, which were observed in the distal part, depending on when the groups of U-axons were divided (Okada, Mizuhira and Nakamura 1976a).     

Enzymatic analysis of individual anterior horn cells in amyotrophic lateral sclerosis and Duchenne muscular dystrophy

Five enzyme activities related to glucose metabolism, i.e. glucose-6-phosphate dehydrogenase (G6PDH), isocitric dehydrogenase (ICDH), citrate synthase (CS), ATP citrate lyase (ACL), and pyruvate dehydrogenase complex (PDC), were estimated in anterior horn cells (AHCs) of spinal cords in amyotrophic lateral sclerosis (ALS) and Duchenne muscular dystrophy (DMD) patients by means of the NADP and CoA cycling methods.In ALS, only CS activity was reduced significantly in AHCs, but that of posterior root ganglion cells (PRGCs) remained within the normal range.In DMD, ICDH, CS, ACL and PDC activities were within normal range. However, G6PDH activities were elevated significantly in 2 out of 3 patients examined.     

Monocularly and binocularly evoked visual responses to patterned half-field stimulation

Visual evoked responses to half-field stimulation were recorded in 18 normal subjects to determine the most suitable techniques for use in detecting hemianopic deficits. The most consistent results were obtained by recording from a transverse array of electrodes using the midfrontal region as reference and stimulating with a reversing checkerboard pattern, each check subtending 50 min of visual angle. Responses recorded from electrodes ipsilateral to the half-field stimulated consisted of an N70, P105, N165 complex. Responses from contralateral electrodes demonstrated the presence of an N100 which divided the P105 seen ipsilaterally into a P85 and a P120. The contralateral predominance of the N100 was further documented by plotting the ratio of the N100 to the immediately preceding P wave. Consideration of the ipsilateral P105 and the contralateral N100 together was found to be more reliable in detecting lateralization of the visual evoked potential than either wave alone. Crossed and uncrossed pathways made equal contributions to the visual evoked potential produced by half-field stimulation. Good correspondence was found between algebraic summation of homonymous monocular hemifield responses when compared to binocular hemifield stimulation. This finding supports the use of binocular hemifield stimulation for the detection of subtle congruous homonymous field defects.     

Autonomic nerve calcification and peripheral neuropathy in olivopontocerebellar atrophy

This report concerns an unusual form of olivopontocerebellar atrophy (OPCA) of adult onset, inherited as an autosomal dominant. We examined 5 patients in one generation and performed neuropathological investigations in 3 of these. The clinical and pathological features were different from those of OPCA types I to V. Apart from olivopontocerebellar degeneration, there was dementia in 4, massive atrophy of the spinal cord in 3, and focal degeneration of the optic nerves in at least 1 case. The most remarkable findings were, however, the involvement of the peripheral nervous system and the abundant intrafascicular calcification in sympathetic nerve fibres and in their ganglia.     

Locus ceruleus lesion and chronic reserpine treatment: Effect on adrenergic and cholinergic receptors in cerebral cortex and hippocampus

Destruction of the central noradrenergic supply to rat cerebral cortex and hippocampus by the local injection of 6-hydroxydopamine into the locus ceruleus produced β-adrenergic supersensitivity but no change in the binding characteristics to α-adrenergic and muscarinic cholinergic receptors in both the cerebral cortex and hippocampus. Chronic reserpine treatment, which caused functional denervation without structural anatomic changes in noradrenergic neurons, produced qualitatively similar results. However, the reserpine-induced β-adrenergic supersensitivity in the cerebral cortex was quantitatively higher than that caused by locus ceruleus lesion. This discrepancy may be due to the presence of presynaptic β-adrenergic receptors that were destroyed by 6-hydroxydopamine but left intact by reserpine.     

Autoradiographic demonstration of retinal projections to the brain stem structures in the rabbit using transneuronal tracing technique with special reference to the retinal projections to the inferior olive

Retinal projections to the brain stem structures in the rabbit were examined autoradiographically using transneuronal tracing technique. Three or four weeks after intraocular injections of tritiated proline and tritiated fucose, significant amounts of silver grains indicating transneuronal labeling of axon terminals were present bilaterally in the visual cortical areas, the dorsal portions of the medial geniculate nucleus, and the suprageniculate nucleus, and contralaterally in the thalamic reticular nucleus, the lateroposterior-pulvinar nuclear complex, the parabigeminal nucleus, the pontine tegmental reticular nucleus of Bechterew, the dorsolateral, lateral, and paramedian pontine nuclei, the pontine reticular formation, and the dorsal cap and beta nucleus of the inferior olive. The label in the pontine regions was probably due to the afferent fibers from the pretectal nuclei and the superior colliculus, and the label in the inferior olive was considered to depend on the uncrossed afferent fibers from the pretectal nuclei and the nuclei of the accessory optic tract.     

Postnatal growth and development of the gerbil brain

The Mongolian gerbil brain undergoes most of its postnatal growth and development during the first 2 weeks of life. At birth the gerbil weighs only 4% of its adult body weight, and the cerebrum and cerebellum are only 24 and 9.9%, respectively, of their adult weights. The cerebrum is more mature at birth than is the cerebellum. At 7 days of age, the cerebrum contains 55.7% of its adult DNA content, whereas the cerebellum has amassed only 17.2% of its adult content. By 21 days of age, cerebral growth is essentially complete, as measured by DNA, RNA, and protein accumulation. Cholesterol content approaches adult values in the cerebrum by 60 days of age. The rate of growth of the cerebrum and rate of RNA accumulation peak at 7 to 8 days of age. The cerebral protein velocity curve reaches its maximum at 10 to 11 days, that of DNA at 12 days, and cholesterol at 17 days of age. Conversely, the cerebellum at 21 days of age has only 65.6% of its adult protein content and 77.4% of its adult weight, although DNA multiplication is nearly complete. Myelination, as measured by accumulation of cholesterol, is not complete until sometime between 90 and 180 days of age. The velocity maxima of the other cerebellar constituents occur at about 12 to 13 days of age.