Computer-assisted system combining image analysis and chromosome dissection.
Explore the source record for details and available documents.
Chromosomes↗
Biomedical subjects
N Leclerc
Publications and source records attributed to N Leclerc.
[Salmonella typhimurium cerebral abscess: rare complication of HIV infection].
Explore the source record for details and available documents.
AIDS-Related Opportunistic Infections↗
Juvenile and mature MAP2 isoforms induce distinct patterns of process outgrowth.
Microtubule-associated protein-2 (MAP2) is the most abundant MAP in neurons, where its distribution is restricted to the somatodendritic compartment. This molecule undergoes developmentally regulated alternative splicing, resulting in at least two isoforms, a juvenile isoform (termed MAP2c) and a mature isoform (MAP2), with greatly different molecular masses. Spodoptera frugiperda (Sf9) cell expression of the juvenile versus the mature MAP2 isoform generates two distinct patterns of process outgrowth. The smaller juvenile isoform induces multiple short thin processes. Mature MAP2 tends to induce single processes that are considerably thicker than those processes induced by juvenile MAP2. We found important differences in the variability of spacing between microtubules and the number of microtubules along the processes induced by MAP2c and mature MAP2. MAP2c showed variability with most microtubules spaced as closely as with tau, but some spaced as far apart as with mature MAP2. Over their length, the mature MAP2 processes demonstrate proximo-distal taper, which corresponds to a narrowing of the spacing between microtubules from 90 nm to 40 nm. Moreover, there is a decreased number of microtubules in mature MAP2-induced processes whereas in tau and MAP2-induced processes, the number of microtubules is constant along the length. Based on these observations, we conclude that MAP2 isoforms can serve as architectural elements by establishing specific morphological features of processes and specific arrangements of their microtubules.
Animals↗
Purkinje cell compartments in the reeler mutant mouse as revealed by Zebrin II and 90-acetylated glycolipid antigen expression.
The cerebellum is organized into a series of parasagittally aligned bands that may be revealed histologically in the adult mouse by largely complementary immunostaining of Purkinje cells sets with the monoclonal antibodies Zebrin II (ZII; antigen:aldolase C) and P-path (PP; antigen:90-acetyl glycolipids). We compared the normal staining pattern using these markers and an antibody to calbindin with that found in the reeler mutants (rl/rl), in which most Purkinje cell migration is halted beneath the cerebellar white matter. The results revealed that Purkinje cells in reeler mutants, despite their ectopic location in large subcortical masses, show a clear tendency to distribute into alternating zones that either stain for Zebrin II or for P-path, with variable transition zones of mixed labeling. However, the estimated number of zones was fewer than in the normal adult cortex: roughly 7-9 zones are revealed per side in the mutant compared with 14 major divisions in wild type mice. These results raise the possibility that neurons destined to express these markers are segregated during their migration and that the final phase of migration into the cortex might involve further splitting or interdigitation between cell sets expressing the two antigens.
Animals↗
Effects of nervous mutation on Purkinje cell compartments defined by Zebrin II and 9-O-acetylated gangliosides expression.
The cerebellum is organized into a series of parasagittally aligned bands which are well delineated in the adult mouse by the largely complementary immunostaining of Purkinje cell groups with the monoclonal antibodies Zebrin II (ZII; antigen: aldolase C) and P-path (antigen: 9-O-acetyl gangliosides). We examined the effect of nervous mutation on compartmental organization using these markers and an antibody to calbindin. In nervous mutant, up to 90% of Purkinje cells die in late postnatal development. The size of the cerebellum is about half that of normal, and caudal lobules appear to decrease in size more than anterior ones. Surviving Purkinje cells corresponded to P-path positive ones that were concentrated in two bilateral bands in the vermis and in medial portions of the hemispheres. Only small numbers of ZII positive cells remained, confirming the report by Wassef et al. with Zebrin I antibody. They were primarily located in caudal lobules IX, X and a portion of lobule IV, paraflocculus and flocculus, and their immunoreactivity was weak compared to that of normal. ZII positive cells are dominant in these caudal lobules, while P-path positive cells dominate in rostral lobules in normal mice, and the similar tendency remains in mutant. Thus, the nervous gene action respects not only sagittal compartments delineated by two antibodies, but also rostro-caudal gradient. The cause of the dominant survival of P-path positive cells awaits future study.
Animals↗
Compartmentation in mammalian cerebellum: Zebrin II and P-path antibodies define three classes of sagittally organized bands of Purkinje cells.
The respective roles of genetic and epigenetic factors in generation of pattern formation in the vertebrate nervous system are still poorly elucidated. The mammalian cerebellum is subdivided in parasagittal modules defined by anatomical, physiological, and biochemical criteria. Immunostaining of adult mouse cerebellum with two monoclonal antibodies, P-path, which recognizes 9-O-acetylated glycolipids, and Zebrin II, which recognizes a 36-kDa protein, reveals three classes of sagittally organized bands of Purkinje cells: two complementary groups distinctly immunoreactive to one antibody but not the other and a third group that contains double-labeled cells. No Purkinje cells could be detected that were unreactive to either antibody. The specific and reproducible topography of these three classes of Purkinje cells may be related to the compartmentation of the cerebellum into developmental genetic modules.
Animals↗
Expression of compartmentation antigen zebrin I in cerebellar transplants.
The mammalian cerebellum is divided into multiple parasagittal compartments as defined by the organization of afferent and efferent projections and by the pattern of expression of several biochemical markers. One such marker is the antigen zebrin I, a 120 kD polypeptide of unknown function that is expressed differentially by a subset of Purkinje cells. Zebrin I+ Purkinje cells are grouped into an array of 14 parasagittal bands interposed by zebrin I- compartments. This Purkinje cell compartmentation corresponds to compartments in the olivocerebellar projection. The afferent axon compartments are present prior to the expression of the mature zebrin I phenotype, thus raising the possibility that differential afferent input regulates the zebrin I phenotype of the target of that input. Lesion studies in the neonate preclude a role for afferent inputs in the regulation of zebrin I expression postnatally, but a prenatal role in commitment still remains open. To explore this possibility, cerebellar anlagen were dissected from embryos at embryonic days 12-15, that is, prior to any contact with afferents, and transplanted ectopically into adult hosts. In the first series of experiments, the grafts were placed into the anterior chamber of the eye, and in the second series, into cavities prepared in the neocortex. Grafts were allowed to mature and then were immunoperoxidase or immunofluorescence stained for zebrin I immunoreactivity. Zebrin I was expressed by grafted Purkinje cells in cortico and in oculo. Double-labelling experiments confirmed that both the zebrin I+ and the zebrin I- phenotypes were present. The zebrin I immunoreactivity revealed that the zebrin I+ Purkinje cells resemble those in situ with an extensive dendritic arborization that extends through the molecular layer perpendicular to the long axes of the folia. In conclusion, the present data suggest that afferent input does not play a role in the determination of the zebrin I phenotype of Purkinje cells.
Animals↗
The compartmentalization of the monkey and rat cerebellar cortex: zebrin I and cytochrome oxidase.
The cerebellar cortex of mammals is composed of parasagittal zones that encompass the afferent inputs, the efferent corticonuclear and corticovestibular projections, and a number of intrinsic molecular markers. One such marker is the polypeptide antigen zebrin I that is recognized by monoclonal antibody (mab) Q113. In rodents, zebrin I immunocytochemistry reveals an array of parasagittal Purkinje cell compartments. In the present study, zebrin I has been used to reveal the molecular heterogeneity of the cerebellar cortex in the squirrel monkey (Saimiri sciureus). As in rodents, zebrin I is Purkinje cell specific in the primate cerebellum and not all Purkinje cells are immunoreactive. Immunocytochemistry on frontal or horizontal sections reveals a system of bands of zebrin I+ cells extending through the vermis of both anterior and posterior lobes. A midline (P1+) band and two more lateral bands (P2+ and P3+) are found in all lobules. The situation in the paravermis and hemispheres is similar, with alternating zebrin I+ and zebrin I- compartments, but the complex lobulation obscures the precise band pattern: it seems probable that 4 additional bands are present in the hemispheres, as in rodents. Comparison of rat and monkey cerebellums suggests that the cortex has expanded in primates by the growth of the same individual bands found in rats rather than by the addition of supplementary compartments. The zebrin I compartmentalization revealed by using mab Q113 is reproducible from individual and thus provides a stable frame of reference that has been used to compare the different chemoarchitectonic patterns found in the cerebellar cortex.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Synaptophysin expression during synaptogenesis in the rat cerebellar cortex.
In order to study the mechanisms of synaptogenesis in the rat cerebellar cortex, a library of monoclonal antibodies has been generated against proteins of the isolated synapse. One recognizes a glycosylated 38 kDa protein that is concentrated in the synaptic vesicle fraction and resembles synaptophysin biochemically in its molecular weight, charge, and pattern of glycosylation. In the adult cerebellar cortex, the antisynaptophysin(mabQ155) immunoreactivity is codistributed with synapses. Immunoreactivity is strongest in the molecular layer where punctate deposits of reaction product outline the Purkinje cell dendrites. Discrete small profiles, consistent with the distribution of basket cell axon terminals, surround the Purkinje cells, and in the granular layer the synaptic glomeruli are intensely stained. There is no immunoreactivity in the white matter axon tracts. Electron microscope immunocytochemistry confirms the synaptic location of the antigen and suggests that the reaction product is associated with synaptic vesicles. Both round and flat vesicle populations are immunoreactive. Antisynaptophysin(mabQ155) has been used to follow synaptogenesis in the developing rat cerebellum. In the newborn rat (P0), despite the paucity of synapses, there is some specific immunoreactivity, especially in the subcortical white matter. Electron microscopy shows that the antigenicity is associated with vesicles within growth cones, filopodia, and immature axon profiles. During development, antisynaptophysin immunoreactivity increases progressively, along with the maturing cell populations, for both the granule cell-Purkinje cell and the mossy fiber-granule cell synapses. Quantitative biochemical analysis confirms the cytochemical results. These data suggest that neuronal growth cones express a synapse-specific antigen before complete morphological synapses are present.
Aging↗
Purkinje cell axon collateral distributions reflect the chemical compartmentation of the rat cerebellar cortex.
Monoclonal antibody mabQ113 has been used to study the distribution of Purkinje cell axon collaterals in the rat cerebellar cortex. MabQ113 recognizes a polypeptide antigen, zebrin I, that is confined to a subset of Purkinje cells. Antigenic Purkinje cells are arranged in parasagittal compartments running throughout the cortex. No other cerebellar cells are immunoreactive. Immunoreactive axon collaterals are confined principally to the infraganglionic plexus with only a few extending into the molecular layer. Three probable target cells for the axon collaterals have been identified: Golgi cells, Lugaro cells, and other Purkinje cells. About 90% of immunoreactive axon collaterals in the anterior lobe are located beneath the mabQ113+ Purkinje cell compartment in which they originate but some do invade the neighboring mabQ113- territory. In the anterior lobe vermis, the distribution of invading mabQ113+ collaterals is not symmetrical, such that the probability of an invading collateral from the P2+ compartment is greater at the medial boundary into P1- than at the lateral into P2-. The distribution of immunoreactive collaterals is consistent with their playing a role in synchronizing the firing of Purkinje cells within the same compartment.
Animals↗
Development of parasagittal zonation in the rat cerebellar cortex: MabQ113 antigenic bands are created postnatally by the suppression of antigen expression in a subset of Purkinje cells.
Monoclonal antibody mabQ113 recognizes a polypeptide antigen that, in the adult cerebellum, is confined to a subset of Purkinje cells that are clustered together to form parasagittal bands interposed by similar nonimmunoreactive bands. The Purkinje cell compartments are congruent with bands of climbing fibers projecting from subregions of the inferior olivary complex (IOC). The array of mabQ113 parasagittal bands appears late in the development of the cortex. Weak mabQ113 immunoreactivity is first seen at postnatal day 6 (P6) in the Purkinje cells of the posterior lobe of the vermis. From the earliest stages there are signs of differential expression of the mabQ113 antigen in clusters of Purkinje cells: four mabQ113+ clusters are clearly present in the posterior lobe of the vermis at P6-P7. Their relation to the adult band display remains uncertain. During the next few days immunoreactivity spreads rostrally throughout the rest of the vermis and laterally to include the Purkinje cells in the hemispheres, until by P12 all the Purkinje cells in the cerebellum are mabQ113+. Nevertheless, signs of the adult band display are seen already in the vermis where the cells destined to become the vermal mabQ113+ bands (P1+, P2+ and P3+) stain more intensely than their neighbours. Following the stage of global mab113 epitope expression, bands are created by the selective suppression of immunoreactivity by Purkinje cells in the P- regions. By P15 the mabQ113+ and mabQ113- bands are clearly differentiated in the vermis and selective staining has begun to appear in the hemispheres also. The band pattern matures gradually during the third and fourth postnatal weeks until the adult appearance is attained by P30. The cerebellar afferent projections were lesioned to explore the interplay of cerebellar input and mabQ113 expression. The olivocerebellar projection was lesioned bilaterally by using 3-acetylpyridine in the adult and unilaterally in the newborn by electrolytic lesion and unilateral inferior cerebellar pedunculectomy. Mossy fibers from the dorsal and ventral spinocerebellar tracts were lesioned surgically both in adults and in newborn and trigeminal projections to the cerebellum were removed in the newborn by unilateral ablation of the spinal trigeminal nucleus. The consequences of total blockage of vibrissal and hindlimb inputs were also explored in both adults and neonates. None of these treatments led to a modification in the pattern of mabQ113 epitope expression.(ABSTRACT TRUNCATED AT 400 WORDS)
Aging↗
Antigenic map of the rat cerebellar cortex: the distribution of parasagittal bands as revealed by monoclonal anti-Purkinje cell antibody mabQ113.
Both anatomical and physiological mapping methods have revealed that the mammalian cerebellar cortex consists of a family of parasagittal bands of cells, each band with its own pattern of afferent and efferent axons. Monoclonal antibody mabQ113 recognizes an unknown polypeptide antigen that is confined to a subset of rat cerebellar Purkinje cells. Immunoreactive cells are arranged into parasagittal bands extending throughout the vermis and hemispheres. Expression of the Q113 epitope by individual Purkinje cells may not be all-or-nothing, since the bands tend to be more strongly stained in the vermis than the hemispheres. The band display is symmetrical about the midline and reproducible from individual to individual. Whole-mount immunocytochemistry and serial reconstruction reveal a median band of mabQ113+ Purkinje cells adjacent to the midline (P1+) and six other positive bands disposed symmetrically at either side (P2+ to P7+). Bands are distinct throughout most of the cortex but tend to fuse ventrally and caudally. There are two sources of interindividual differences. Firstly, most animals express supernumerary "satellite" bands in the vermis. Satellite bands are usually only one cell wide, are not bilaterally symmetrical, and differ in position and number from individual to individual. Secondly, the precise position of an individual band can differ, perhaps according to the variable cortical lobulation, for example, the position of P4+ in lobules VIII/IX and P6+ in lobule VII. While a scheme of parasagittal bands is a good description of the vermian organization, the distribution of mabQ113+ and mabQ113- Purkinje cells in the hemispheres may be better described as a checkerboard of antigenic patches.
Animals↗
Monoclonal antibodies reveal the global organization of the cerebellar cortex.
Electrophysiological mapping of the rat cerebellar cortex has revealed an elaborate functional somatotopy that tract tracing procedures have shown to correlate with specific patterns of afferent and efferent connectivity that encompass the cerebellum as a whole. In contrast, most anatomical and biochemical procedures suggest that the cerebellar cortex is remarkably uniform. To unmask covert molecular heterogeneity underlying the functional map, it is appropriate to use monoclonal antibody technology to search for antigenic epitopes whose cerebellar distribution reflects or encodes the positional information. Given that no preconditions can be set on the biochemical nature of the putative epitopes, a shotgun approach to immunization and screening is required. The construction of monoclonal antibodies and screening for specificities that reveal positional information is discussed with examples from an anti-cerebellar antibody library.
Antibodies, Monoclonal↗
A monoclonal antibody to mammalian neurofilament protein stains somata and dendrites in gymnotid fish.
Monoclonal antibody N210 (mabN210) recognizes the 210 kdalton neurofilament protein in mammals and gives characteristic immunocytochemical staining of neurofilament-rich processes. For example, in the cerebellum it recognizes myelinated axons and the calyx formed by basket cell axon collaterals. The distribution of mabN210 immunoreactivity was studied in the gymnotid brain (Apteronotus albifrons). In contrast to the mammalian distribution, mabN210 immunoreactivity was not found in most axons of the gymnotid brain. Instead, deposits of reaction product were present in the somata and dendrites of most neurons and were especially dense in those neurons with extensive dendritic trees, the Purkinje cells, pyramidal cells of the electrosensory lateral line lobe, the crest cells of the nucleus medialis and the pyramidal cells of the tectum. Electrosensory lateral line lobe pyramidal cells are known to contain few, if any, neurofilaments in their dendrites. Western blots of whole gymnotid brain proteins demonstrated that mabN210 recognizes two polypeptides apparent molecular weights 60 and 19 kdaltons. These proteins are thus antigenically similar to neurofilament protein and their expression in the gymnotid brain may be related to the peculiar dendritic branching pattern of Purkinje cells and similar cell types.
Afferent Pathways↗
Immunocytochemical demonstration of topographic ordering of Purkinje cell axon terminals in the fastigial nuclei of the rat.
We have used a monoclonal antibody, mabQ113, which selectively stains a subset of cerebellar Purkinje cells, to study the topography of the corticonuclear projection of the median vermis to the fastigial nuclei in the rat. The fastigial projection zone contains both mabQ113+ and mabQ113- Purkinje cells grouped into parasagittal bands. The immunoreactivity extends throughout the Purkinje cell including the axon terminals and thus it is possible to investigate the topographic distribution of mabQ113+ terminals. In the fastigial nuclei the target cells receiving mabQ113+ axon terminals are concentrated in the caudal pole. In the rostral pole, cells receive anti-GAD+ terminals but not mabQ113+ terminals. There is no gradient in anti-GAD staining. Double-labelling experiments with mabQ113, anti-GAD, and an antisynaptic antibody mabQ155 suggest that there is little or no mixing of mabQ113+ and mabQ113- Purkinje cell terminals on the same target neuron.
Animals↗
Focal axonal swellings in rat cerebellar Purkinje cells during normal development.
Focal axonal swellings are characteristic of a wide range of neuropathies. Three neuron-specific monoclonal antibodies have been used to identify focal axonal swellings in the normal developing rat cerebellar cortex. Between 7 and 15 days postnatal, swellings are a common feature of the granular layer and white matter tracts. Using a Purkinje cell-specific antibody, the majority of swellings were shown to occur in Purkinje cell axons. Focal axonal swellings therefore seem to be a normal adjunct of Purkinje cell maturation.
Animals↗
Monoclonal antibodies reveal sagittal banding in the rodent cerebellar cortex.
We have produced two monoclonal antibodies against polypeptide-associated antigens of developing rat cerebellum. One antibody recognizes an antigen associated with synaptic vesicles and another binds to a polypeptide which is restricted to the cytoplasm of a subset of cerebellar Purkinje cells. Both antibodies reveal the biochemical differentiation of the rodent cerebellar cortex into antigenically distinct sagittal zones.
Animals↗
Basket cell development in the normal and hypothyroid rat cerebellar cortex studied with a monoclonal anti-neurofilament antibody.
We have used a monoclonal antibody against an antigenic determinant of the 210-kdalton neurofilament protein to study basket cell maturation in rat cerebellar cortex. Neurofilament immunoreactivity first appears in basket cells at postnatal day 12 and mature axonal "pinceaux" are present at postnatal day 17. There are large differences in the rate of maturation from lobe to lobe which do not fully correspond to the rate of Purkinje cell differentiation. In hypothyroid rats the expression of the neurofilament antigen by basket cells is almost completely suppressed.
Animals↗