Biomedical subjects

C H Berthold

Publications and source records attributed to C H Berthold.

At least 19 recordsLinked to original sources

PubMed1995

Axonal constriction at Ranvier's node increases during development.

We have studied the ratio between the nodal and the internodal diameter (the dn/d(in) ratio) of large myelinated axons in the L7 ventral spinal root of the cat during pre- and postnatal development using light and electron microscopy. A substantial nodal constriction, dn/d(in) = 0.6, was found at the beginning of myelination, about 2 weeks before birth. The ratio decreased during the subsequent 10 weeks and approached the adult value of 0.47 (SE 0.01, N = 45) in the 8 weeks old kitten. The observations are discussed with respect to the maturation of the nodal region and to our earlier idea that the constricted nodal axon segments of large peripheral myelinated nerve fibres of adult cats and kittens 2 months and more of age are sites capable of interacting with and perhaps even controlling the passage of axonally transported materials.

Age Factors↗

PubMed1995

Mice lacking glial fibrillary acidic protein display astrocytes devoid of intermediate filaments but develop and reproduce normally.

Glial fibrillary acidic protein (GFAP) is the main component of the intermediate filaments in cells of astroglial lineage, including astrocytes in the CNS, nonmyelin forming Schwann cells and enteric glia. To address the function of GFAP in vivo, we have disrupted the GFAP gene in mice via targeted mutation in embryonic stem cells. Mice lacking GFAP developed normally, reached adulthood and reproduced. We did not find any abnormalities in the histological architecture of the CNS, in their behavior, motility, memory, blood-brain barrier function, myenteric plexi histology or intestinal peristaltic movement. Comparisons between GFAP and S-100 immunohistochemical staining patterns in the hippocampus of wild-type and mutant mice suggested a normal abundance of astrocytes in GFAP-negative mice, however, in contrast to wild-types, GFAP-negative astrocytes of the hippocampus and in the white matter of the spinal cord were completely lacking intermediate filaments. This shows that the loss of GFAP intermediate filaments is not compensated for by the up-regulation of other intermediate filament proteins, such as vimentin. The GFAP-negative mice displayed post-traumatic reactive gliosis, which suggests that GFAP up-regulation, a hallmark of reactive gliosis, is not an obligatory requirement for this process.

Animals↗

PubMed1995

Automated correction of linear deformation due to sectioning in serial micrographs.

This paper describes an objective and automatic method for detection and correction of sectioning deformations in digitized micrographs, as well as an evaluation of the method applied to light and electron microscopic images of semi-thin and ultra-thin serial sections from brain cortex. The detection is based on matching of image subregions and the deformation model is bi-linear, i.e. two first-order polynomials are used for modelling compression/expansion in perpendicular directions. The procedure is applicable to prealigned serial two-dimensional sections and is primarily aimed at three-dimensional reconstruction of tissue samples consisting of a large number of cells with random distribution and morphology.

Animals↗

PubMed1994

A method for 2D reconstruction of intracellularly labeled neurons from sequential sections.

A technique for 2D reconstruction of intracellularly labeled neurons from sequential sections is described. The system consists of a Charged Coupled Device-camera mounted on a microscope, a videomixer and a IBM-compatible PC with a framegrabber. The neurons (interneurons from the spinal cord of the cat) were labeled iontophoretically by horshradish peroxidase and subsequently cut in 60 microns sections. The sections were aligned using the video mixer by fitting the cut dendrites and axon from one section with their counterparts in the following section. The images were then digitized in the PC where they were fused to create a superimposed picture of the aligned parts of the neuron; a 2D reconstruction was created.

Animals↗

PubMed1994

Dimensions of individual alpha and gamma motor fibres in the ventral funiculus of the cat spinal cord.

Using light and electron microscopy, axon diameter, myelin sheath thickness (measured as number of myelin lamellae) and internodal length of alpha and gamma motor axons of the L7 ventral root and spinal cord segment were investigated in serial cross-sections. The CNS internodes of the alpha motor fibres had, on average, an axon diameter of 8.6 microns, 105 myelin lamellae and a length of about 560 microns. The CNS internodes of the gamma motor fibres had, on average, an axon diameter of 3.4 microns, 66 myelin lamellae and a length of about 440 microns. Axon diameter at the nodes of Ranvier was 30-40% of the internodal axon diameter. Axon diameter, number of myelin lamellae and internodal length varied considerably between consecutive internodes. Statistical analysis showed no systematic increases or decreases. Regression analyses of the scatter plots of the number of myelin lamellae and internodal length against axon diameter showed large variations and correlation coefficients of r < 0.50. In conformity with ventral root (PNS) internodes (Nilsson & Berthold, 1988) the plotting of intrafunicular (CNS) internodal myelin volume against internodal axon mantle area showed linear correlations with correlation coefficients of r > 0.90. The mean axon diameter of the investigated CNS internodes was similar to, the mean number of myelin lamellae somewhat lower than, and the mean internodal length considerably shorter than that of internodes of axons of the L7 ventral root (Nilsson & Berthold, 1988). In contrast to the ventral root, the intrafunicular alpha motor fibres had higher g values (axon diameter/fibre diameter value) and lower il/d ratios (internodal length/axon diameter ratio) than is considered optimal for conduction. We consider that these deviations from the theoretical optimum are not large enough to impair the conduction properties of the CNS parts of the motor axons in a significant way.

Animals↗

PubMed1993

Axoplasmic organelles at nodes of Ranvier. I. Occurrence and distribution in large myelinated spinal root axons of the adult cat.

Using light microscopy (LM) and electron microscopy (EM) we have examined the occurrence and distribution of axoplasmic organelles in large myelinated nerve fibres of the L7 ventral and dorsal spinal roots of the cat with special reference to the paranode-node-paranode (pnp)-regions. Ninety-eight percent of the 550 Toluidine Blue-stained paranode-node-paranode-regions examined in the light microscope contained dark-blue bodies accumulated distal to the midlevel of the paranode-node-paranode-region. Further, a veil of Toluidine Blue positive material was observed in about 50% of the paranode-node paranode-regions. In about 25% of these paranode-node-paranode-regions the veil lay distal to the midlevel of the paranode-node-paranode-region and in the remainder it lay proximally. Electron microscopy suggested that the ultrastructural equivalents of the dark-blue bodies and of the veil were dense lamellar bodies and a diffuse granular material, respectively. Our calculations indicate that from 70% to more than 90% of some organelles (dense lamellar bodies, multivesicular bodies and vesiculo-tubular membranous organelles) present in an axon are accumulated in the paranode-node-paranode-regions. The occurrence of these organelles in the individual paranode-node-paranode-regions varied within wide limits also in adjacent fibres. The dense lamellar and multivesicular bodies dominated the distal part of the paranode-node-paranode-regions while the vesiculo-tubular membranous organelles dominated the proximal part, i.e. the organelles showed a mutual proximo-distal segregation with reference to the midlevel of the paranode-node-paranode-region. Of seventeen paranode-node-paranode-regions analyzed ultrastructurally, seven were classified as 'fully segregated', that is 67% or more of the lamellar and multivescular bodies, present in the whole paranode-node-paranode-region, lay distal to the mid-level, and 67% or more of the vesiculo-tubular membranous organelles lay proximal to it.

Animals↗

PubMed1993

Axoplasmic organelles at nodes of Ranvier. II. Occurrence and distribution in large myelinated spinal cord axons of the adult cat.

The occurrence and distribution of axoplasmic organelles in large myelinated axons of the ventral, the lateral and the dorsal funiculi of L7 spinal cord segments of the cat have been studied using electron microscopy (EM). Most organelles were found to be concentrated to the paranode-node-paranode (pnp)-regions and they showed their highest relative concentration in the constricted part of these regions, i.e. at the nodes of Ranvier. In the paranode-node-paranode-regions of the lateral and dorsal funiculi, large dense bodies predominated distal to the nodal mid-level and vesiculo-tubular membranous organelles proximal to it. This pattern of organelle distribution, a proximo-distal (with reference to the neuron soma) segregation of the organelles, was only faintly indicated in the paranode-node-paranode-regions of the alpha motor axons of the ventral funiculus. These paranode-node-paranode-regions were, apart from a weak proximo-distal segregation of a few organelles, characterized by deposits of electron dense granules and clusters of large round mitochondria. We conclude that there are two types of organelle accumulation and distribution in the paranode-node-paranode-regions of large spinal cord nerve fibres of the cat. One type is found in the lateral and dorsal funiculi, i.e. in axons with terminal (synaptic) fields inside the blood-brain-barrier. The other type is found in the alpha motor axons of the ventral funiculus, i.e. in axons with their terminal field in the PNS and thus outside the blood-brain barrier. It should be noted that retrogradely transported material in the alpha motor axons has passed through a long sequence of paranode-node-paranode-regions equipped with Schwann cells before it reaches the CNS, while material transported retrogradely in the axons of the dorsal and lateral funiculi has not. The following discussion includes a comparison of the organelle accumulation and distribution in these two types of CNS paranode-node-paranode-regions with the organelle accumulation and distribution observed in the paranode-node-paranode-regions of PNS axons.

Animals↗

PubMed1992

Metabolic relationships between proteins of myelin and paranodally shedded, partially degraded myelin fragments in the rabbit CNS.

The "close-to-node" regions of myelinated nerve fibres, i.e., the paranodal end segments, are generally thought to be sites of high metabolic activity and myelin sheath turnover. Data on turnover rates of individual myelin constituents are conflicting but there exists a common belief that myelin is metabolized as independent molecules rather than as a unit. The occurrence of paranodal Marchi-positive bodies, with morphological and biochemical properties consistent with partially degraded myelin, prompted us to examine the temporal dynamics of the incorporation of radioactive precursor label in the major proteins of myelin and the Marchi-positive bodies. 3H-leucine was administered intrathecally in adult rabbits. After various survival times, the spinal cord was subfractionated by ultracentrifugation in a discontinuous two-step 0.32 M/0.85 M sucrose gradient. Myelin was collected from the interface and a floating fraction, heavily enriched in Marchi-positive bodies, was recovered on top of the 0.32 M sucrose. By scintillation counting and by gel fluorography combined with immunoblotting, a gradual appearance with time of partially degraded peptides of myelin-associated protein and 2',3'-cyclic nucleotide 3'-phosphodiesterase was seen in the floating fraction but not in myelin. The temporal dynamics of the specific activities of these two proteins and myelin-basic protein and proteolipid protein were consistent with a typical source-product relationship between myelin and the material in the floating fraction. In conjunction with earlier morphological and biochemical findings, these data may suggest that Marchi-positive bodies appear as a consequence of myelin catabolism.

Animals↗

PubMed1992

Computer-assisted realignment of light micrograph images from consecutive section series of cat cerebral cortex.

Two computer- and image-analysis-based procedures for realignment of images of consecutive light micrographs of nerve tissue (cat motor cortex) have been developed. One procedure (CENT) was interactive, employing the subjective overall 'best fit' of two images to each other and the other (AUTOCENT) was based on an automatic comparison of two images (image thresholding and binary comparison). Images of light micrographs of several hundred consecutive sections were realigned using the interactive and the automatic methods. The interactive procedure was easy to use; realignment of an image took only a few minutes in the hands of an experienced operator and should be easy to implement on commercially available smaller computers like PCs and workstations, but the realignment was possibly disturbed by the operator's subjective expectancy of regular forms. The automatic procedure realigned 1-4 images per hour, might be implemented on smaller computers if the programming is made more efficient, was probably objective and restored the 'true position of images'.

Animals↗

PubMed1991

Lysosomal activity at nodes of Ranvier in dorsal column and dorsal root axons of the cat after injection of horseradish peroxidase in the dorsal column nuclei.

The occurrence of acid phosphatase (AcPase)-positive bodies, i.e. lysosomes, in dorsal column and dorsal root axons of the spinal cord segments C8 and L7 in adult cats was analyzed by light and electron cytochemical methods after injection of horseradish peroxidase (HRP) in the dorsal column nuclei. Axonal lysosomes were, with few exceptions, concentrated at the nodes of Ranvier. We found no changes in nodal occurrence and distribution of lysosomes in axons of the HRP-injected sides, as compared to axons of the uninjected sides or of animals not exposed to HRP. Axonal lysosomes were very rare in the dorsal columns, where the frequency of nodes containing light microscopically detectable AcPase-positive bodies was 0-5% at the HRP-injected sides, 0-6% at the contralateral sides, and 0-3% in control animals. The corresponding values in the cervical and lumbar dorsal roots were 6-23%, 9-20%, 10-12% and 19-37%, 21-40%, 26-43%, respectively. In view of our recent observations in alpha-motor neurons, the results point at a noteworthy difference in local degradative ability between dorsal column axons and alpha-motor axons, the latter being able to accumulate intramuscularly injected and retrogradely transported HRP at their PNS nodes of Ranvier for 48-60 h, during which period the axoplasmic AcPase activity/concentration increases at some nodes. Such a degradative activity, which could protect the motor neurons by restricting axoplasmic transport of exogenous materials imbibed by their axon terminals outside the CNS, may not be of the same significance for neurons, e.g. dorsal root ganglion neurons, the axon terminals of which are located within the CNS.

Acid Phosphatase↗

PubMed1991

Lysosomal activity in developing cat alpha-motor axons under normal conditions and during retrograde axonal transport of horseradish peroxidase.

The occurrence of acid phosphatase (AcPase)-positive bodies, i.e., lysosomes, in lumbosacral alpha-motor axons of kittens, 0-16 weeks of age, was analyzed by light and electron cytochemical methods under normal conditions and after intramuscular injection of horseradish peroxidase (HRP). Axonal lysosomes were rare early postnatally. In 3-week-old animals, a few AcPase-positive bodies appeared in the axoplasm at some nodes of Ranvier in the peripheral nervous system (PNS) and internodally in the intrafunicular motor axon parts within the central nervous system (CNS). From 6 weeks postnatally, a nodal concentration of AcPase-positive bodies was also noted in the CNS. The number of AcPase-positive bodies continued to increase gradually in the course of neuronal maturation. In 16-week-old animals, axonal AcPase activity was still at considerably lower levels than at adult stages. At all ages, acid hydrolase-containing organelles were most commonly found at ventral root nodes. After injection of HRP in the medial gastrocnemius muscle, accumulations of AcPase-positive bodies were seen in the axoplasm at some PNS nodes of the HRP-injected sides of kittens aged 8, 12, and 16 weeks. Incubation for demonstration of both HRP and AcPase activity showed that some organelles at HRP-transporting nodes contained both types of reaction product. The nodal AcPase activity in the intrafunicular, CNS parts of alpha-motor axons of the HRP-exposed sides did not differ from that of the contralateral, uninjected sides. In view of our previous observations in alpha-motor neurons of adult cats in which a lysosome-mediated degradation of axonally transported materials may take place at PNS nodes of Ranvier, the present study illuminates possible differences in the ability to interfere with axonal transport between developing and mature neurons. The infrequent presence of lysosomes in developing alpha-motor axons and the implied disability of their nodal regions to interfere with axonally transported constituents in a way similar to that seen in adult animals may be of significance in that trophic and chemical signals can pass unhindered between the periphery and perikaryon. However, this could also have negative consequences for the vulnerable immature neuron in that various materials retrieved at the axon terminals outside the CNS are permitted a more-or-less free access to the perikaryon.

Aging↗

PubMed1991

Axon-Schwann cell networks are regular components of nodal regions in normal large nerve fibres of cat spinal roots.

The paranodal occurrence of axon-Schwann cell networks (ASNs), which are entities assumed to take part in the removal of degenerate axonal material, was examined quantitatively by electron microscopical serial section analysis in normal cat ventral and dorsal spinal roots. In nerve fibres greater than or equal to 10 microns in diameter 88% of the nodal regions in the ventral roots and 97% in the dorsal roots showed ASN complexes, which especially in the ventral roots often consisted of many segregated axoplasmic portions. The corresponding frequencies in fibres less than 10 microns were 28% and 62% in the ventral and the dorsal roots, respectively. ASN complexes were rare in fibres less than 5 microns. The results show that the ASN is a part of the normal paranodal architecture in large myelinated nerve fibres. The ASN occurrence seems to differ with neurone type.

Animals↗

PubMed1991

Cellular relationships of paranodal Marchi-positive bodies studied with monoclonal antibodies against partially degraded CNS myelin fragments.

Histochemical and electron microscopical studies have shown that Marchi-positive bodies in the normal mammalian CNS are associated with paranodal regions and it has been proposed that the formation of Marchi-positive bodies represents a step in the catabolic events of normal myelin turnover. After a two-step density gradient ultracentrifugation a light 'floating fraction' highly enriched in these structures can be collected and in the present study two monoclonal antibodies, FC4 and 3B5, were produced against proteins present in the floating fraction but absent from the myelin fraction. Immunohistochemical studies showed that these antibodies bound preferentially to the PNS-CNS transitional region and the glia limitans. Double-staining experiments demonstrated an extensive overlap in these regions with cells stained by antibodies against the astrocyte marker glial fibrillary acidic protein. Centrally in the white matter, FC4 and 3B5 mainly stained cells which also stained with the microglia lectin marker Bandeiraea simplicifolia isolectin B4. Three-dimensional reconstructions made from confocal microscopic scans showed that FC4/3B5-positive cells in the white matter extend processes enveloping paranodal Marchi-positive bodies and nodes of Ranvier. It is suggested that astrocyte-like and microglia-like cells both are participants in paranodal myelin turnover and that a division of labour with respect to, for example, protein degradation and immunological functions, may be present between the two cell types.

Animals↗

PubMed1990

Lysosomal activity at nodes of Ranvier during retrograde axonal transport of horseradish peroxidase in alpha-motor neurons of the cat.

Lysosomal activity at nodes of Ranvier of feline hindlimb alpha-motor neurons was examined by light and electron microscopical acid phosphatase (AcPase) histochemistry during retrograde axonal transport of intramuscularly injected horseradish peroxidase (HRP). Several nodes along the PNS parts of the alpha-motor axons of the HRP-injected side showed accumulations of AcPase-positive bodies in the constricted nodal axon segment and the adjacent paranodal axoplasm. Such lysosomal accumulations were most prominent in the ventral root and differed in number and intensity depending on survival time after the HRP injection. At nodes showing high AcPase activity the axoplasm proximal to the nodal midlevel was occupied by many small, AcPase-positive, vesiculotubular profiles. Larger AcPase-positive bodies were mainly situated distal to the nodal midlevel. Double incubation for demonstration of both HRP and AcPase activity showed similar accumulations of AcPase-positive bodies at some of the HRP-transporting nodes. The AcPase activity differed considerably between nodes exhibiting comparable levels of HRP-positivity. Many of the AcPase-positive bodies also contained HRP reaction product. At some HRP-positive nodes the number of AcPase-positive bodies situated in the paranodal axon-Schwann cell network was elevated when compared to nodes of the contralateral, control side. In contrast to the PNS nodes, the nodal occurrence and distribution of lysosomes in the CNS part of alpha-motor axons seemed not to be affected by HRP transport. These observations support our previous proposal that nodes of Ranvier in the PNS parts of alpha-motor axons, in contrast to their CNS nodes, possess an ability to control passage of and initiate lysosomal degradation of axonally transported substances. Such an ability may provide a protective function to the motor neuron by restricting the intraneuronal transport of materials imbibed by the axon terminals outside the CNS.

Acid Phosphatase↗

PubMed1989

Marchi-positive myelinoid bodies at the transition between the central and the peripheral nervous system in some vertebrates.

The CNS-PNS (central nervous system-peripheral nervous system) transitional region of cranial and spinal nerve roots in some vertebrate species was analysed with respect to the occurrence and the distribution of myelinoid Marchi-positive bodies. Both cranial and spinal nerve roots contained more Marchi-positive bodies in their CNS than in their PNS segments. An accumulation of Marchi-positive bodies was usually noted just central to the CNS-PNS borderline. Comparisons between calibre spectra and Marchi index in the cat revealed a particularly high number of Marchi-positive bodies in nerve roots with a high content of myelinated fibres with diameters greater than or equal to 5 microns. Marchi-positive bodies were absent in CNS tissue lacking myelinated nerve fibres. CNS borderline internodes measuring between 200 and 300 microns in length were noted in fibres as thick as 15 microns in feline S1 ventral and dorsal roots. The general picture was similar in all analysed species. Noteworthy however, was the small difference in number of Marchi-positive bodies between CNS and PNS tissue in Xenopus. The chicken contained many myelinoid bodies of similar size and texture as the Marchi-positive bodies but without the Marchi-positive staining properties. The results show that normally occurring Marchi-positive bodies in the CNS are more numerous along paranodal segments than along mid-internodal segments of myelinated nerve fibres and thus support the hypothesis that Marchi-positive bodies are preferentially derived from paranodal myelin.

Animals↗

PubMed1988

Peroxidase activity at CNS nodes of Ranvier and in initial axon segments of lumbosacral alpha-motoneurons after intramuscular administration of horseradish peroxidase.

The occurrence of peroxidase activity in central (CNS) and peripheral nervous system (PNS) parts of alpha-motor axons was studied by light and electron microscopy in adult cats after injection of horseradish peroxidase (HRP) into the medial gastrocnemius muscle. The intrafunicular parts of the axons were virtually free of HRP-positive bodies except at a few nodes of Ranvier. Most of these nodes were weakly HRP-positive and contained, irrespective of a survival time between 25 and 48 h, only a few HRP-positive bodies randomly scattered in the nodal axoplasm. In contrast to this and as described elsewhere (J. Neurocytol., 15 [1986] 253-260), the nodal regions of alpha-motor axons at the level of the ventral root showed strong and characteristic accumulations of HRP-activity. The initial axon segments and adjoining axonal parts contained many HRP-positive bodies. We conclude that the CNS and the PNS parts of an alpha-motor axon differ with regard to the way nodal regions interact with retrogradely transported HRP. Possible mechanisms behind this difference are discussed.

Animals↗

PubMed1988

Aspects of the protein and the lipid composition of myelinoid Marchi-positive bodies from mammalian spinal cord.

The fraction floating on 0.32 M sucrose was isolated from normal mammalian spinal cord and analyzed with regard to protein and lipid composition. Comparisons were made with the myelin fraction isolated from the same spinal cord. A close relationship between the two fractions was indicated by a similar protein banding on SDS-polyacrylamide gel electrophoresis. The relative amounts of various proteins however were different and some high molecular weight proteins appeared unique to the floating fraction. The phospho- and galactolipid patterns, as revealed by thin-layer chromatography, were similar in the floating and the myelin fractions. The proportion of hydrophobic lipids, such as sterols and isoprenyl derivatives, was higher in the floating fraction. Bands co-migrating with cholesterol esters were detected only in the floating fraction from guinea pigs. Marchi-positive material of possible paranodal origin is enriched in the floating fraction. The present findings of a biochemical composition of the floating fraction closely resembling that of myelin is in line with the view that myelin turnover includes a step of degradation localized to the paranodal regions.

Animals↗

PubMed1988

Acid phosphatase activity at nodes of Ranvier in alpha-motor and dorsal root ganglion neurons of the cat.

Acid phosphatase (AcPase) activity in feline alpha-motor and dorsal root ganglion (DRG) neurons was analysed histochemically by light and electron microscopy. The occurrence and distribution of the AcPase activity expressed within the axon differed depending on neuron type and distance from the cell body. Both in alpha-motor and DRG neurons, AcPase-positive bodies of various morphological categories were observed mainly at nodes of Ranvier, where they were more frequent distal than proximal to the nodal midlevel. In the peripherally located processes of both neuron types, most of the larger AcPase-positive bodies were associated with the paranodal axon-Schwann cell network. In the centrally located processes the AcPase-positive bodies were situated in the constricted axon segment and the adjacent paranodal axoplasm. Both in motor and DRG axons, AcPase-positive bodies were more frequent at the spinal root level than at a level central to the PNS-CNS borderline. The observations indicate that lysosomes (i.e. AcPase-positive bodies) constitute part of the intra-axonal system of organelles in normal, large, myelinated alpha-motor and DRG axons of the cat. Lysosome-mediated degradation of retrogradely transported endogenous and exogenous materials may be extensive in normal peripherally directed neuronal processes. The study also suggests a difference between PNS and CNS parts of the same axon with regard to the local turnover of lysosomal organelles.

Acid Phosphatase↗