Biomedical subjects

M R Celio

Publications and source records attributed to M R Celio.

At least 19 recordsLinked to original sources

PubMed1999

Phosphacan immunoreactivity is associated with perineuronal nets around parvalbumin-expressing neurones.

A special feature of the extracellular matrix in adult brains of various species is the concentration of certain components around different sub-populations of neurones, giving rise to net-like structures termed perineuronal nets. Recently, some of these components have been identified but the function of these nets has yet to be resolved. Using immunofluorescence microscopy, we report here that phosphacan, a chondroitin sulphate proteoglycan, is an additional component of Wisteria floribunda labelled perineuronal nets surrounding parvalbumin-expressing neurones in rat cerebral cortex. Glycoproteins such as tenascin-C and -R have been identified in perineuronal nets and the present detection of phosphacan immunoreactivity in the same entity is of potential physiological importance because of their previously described interactions.

Animals↗

PubMed1999

Prolonged contraction-relaxation cycle of fast-twitch muscles in parvalbumin knockout mice.

The calcium-binding protein parvalbumin (PV) occurs at high concentrations in fast-contracting vertebrate muscle fibers. Its putative role in facilitating the rapid relaxation of mammalian fast-twitch muscle fibers by acting as a temporary buffer for Ca2+ is still controversial. We generated knockout mice for PV (PV -/-) and compared the Ca2+ transients and the dynamics of contraction of their muscles with those from heterozygous (PV +/-) and wild-type (WT) mice. In the muscles of PV-deficient mice, the decay of intracellular Ca2+ concentration ([Ca2+]i) after 20-ms stimulation was slower compared with WT mice and led to a prolongation of the time required to attain peak twitch tension and to an extension of the half-relaxation time. The integral [Ca2+]i in muscle fibers of PV -/- mice was higher and consequently the force generated during a single twitch was approximately 40% greater than in PV +/- and WT animals. Acceleration of the contraction-relaxation cycle of fast-twitch muscle fibers by PV may confer an advantage in the performance of rapid, phasic movements.

Animals↗

PubMed1998

Perineuronal nets: past and present.

Golgi ranked the peripheral reticulum--which adheres intimately to nerve cell surfaces--alongside the intracellular reticulum, or Golgi apparatus,which immortalized his name. At first dismissed as an artefact of capricious staining techniques, this peripheral reticulum, or perineuronal net, is now recognized as a genuine entity in neurocytology. It represents a complex of extracellular matrix molecules interposed between the meshwork of glial processes, from which they are indistinguishable, and nerve-cell surfaces. In no other branch of neuroscience has the waxing and waning of interest in any morphological entity been so pronounced as in the case of the perineuronal net. This review traces the history of this enigmatic structure from its conception to the present time, brings to light the keen observational powers of morphologists at the turn of the century and reveals how their sagacious forethought anticipated current thinking on the role of perineuronal nets.

Animals↗

PubMed1998

The onset of parvalbumin-expression in interneurons of the rat parietal cortex depends upon extrinsic factor(s).

Parvalbumin (PV) belongs to the large family of EF-hand calcium-binding proteins and is an excellent marker for a subpopulation of GABAergic neocortical interneurons. During cortical development, PV first appears on postnatal day (P)8, in the infragranular layers; after P14, it also becomes apparent within the supragranular layers. However, nothing is known about the factors controlling its expression, which could involve functional activity, neuronal connectivity and/or neurotrophic factors. It being difficult to manipulate these parameters in vivo, their role may be more readily assessed in organotypic cultures, which are deprived of their subcortical afferents and efferents, and hence of subcortically derived neurotrophic factors and extrinsic functional activity. We prepared slices of the rat brain on P3, P5, P7 and P9, maintained them in culture for 2-5 weeks, and compared the temporal and spatial distribution pattern of PV-immunoreactivity within these slices with the in vivo situation. We found, first, that during late postnatal in vivo development and ageing, the number of PV-immunoreactive neurons in the parietal cortex decreases significantly, and second, that the expression of PV-immunoreactivity in the parietal cortex was markedly influenced by the phase of postnatal development at which slice cultures were explanted. In those removed on P7 and P9, the number of PV-immunoreactive cells, as well as the temporal and spatial distribution pattern of PV-immunoreactivity corresponded to the in vivo situation, but in explants obtained on P3 or P5, PV-immunoreactivity remained confined to layer V of the cortex, reminiscent of the expression profile manifested at the end of the second postnatal week in vivo. Also, the number of PV-immunoreactive cells in these cultures was significantly lower than in explants at the later stages. Our results indicate that the onset of PV-expression in the parietal cortex depends upon extrinsic cortical factors subsisting prior to P7. Once the production of this protein has been initiated, such influences are no longer required.

Animals↗

PubMed1998

McAB 300 antibody against calbindin D-28K is a glial marker in the teleost brain.

The monoclonal antibody McAB 300 against calbindin D-28 k demonstrated an immunostaining pattern in the teleost brain completely different to those obtained in other classes of vertebrates, or after using in the teleost brain other monoclonal and polyclonal antibodies raised against the same protein. Although calbindin D-28k is considered a neuronal marker, McAB 300 specifically stained glial cells throughout the brains of teleosts, with the only exception of some retinal amacrine and horizontal cells.

Animals↗

PubMed1997

Relationship between oligodendrocytes and axons.

Oligodendrocytes can myelinate a variable number of axons in their surroundings; however, the mechanisms underlying axon-oligodendrocyte associations are unknown. We tested the hypothesis that single oligodendrocytes exclusively myelinate axons belonging to the same functional system. Carbocyanine dyes (DiI, FAST DiI) were applied to the sternomastoid muscle of the rat and allowed to transport retrogradely for 4 weeks within the motor axons. Using fluorescence microscopy and iontophoretic injection of Lucifer Yellow (LY), oligodendrocytes were injected in the proximity of retrogradely carbocyanine dye-labeled axons. Using dual channel confocal laser scanning microscopy (CLSM), the three-dimensional relationship between axons and glia was studied. The data indicate that a single oligodendrocyte can myelinate retrogradely labeled axons concomitantly with other, unlabeled axons belonging to apparently different functional systems.

Animals↗

PubMed1997

Immunolocalization of the plasma membrane Ca2+ pump isoforms in the rat brain.

Ca2+ homeostasis in nerve cells is dependent on at least three mechanisms: Ca2+ channels, calcium-binding proteins and Ca2+ exchangers/pumps. Only limited information is available on the regional/cellular distribution of these Ca2+-regulating systems in the brain. The distribution of three of the isoforms of one of the systems, plasma membrane Ca2+-ATPase (PMCA), was analyzed in this study. Using antibodies against epitopes specific for each isoform, a map of the distribution of the pump in the whole brain was produced. The pump was mainly expressed in neurons and was apparently absent from glia cells. Isoform 1 was ubiquitous and occurred in varying, but always significant, concentrations in almost all nerve cells. Isoform 2 was abundant in cerebellar Purkinje cells but less concentrated in other brain regions. Isoform 3 had a predominantly extra neuronal location, e.g. it was abundant in the choroid plexuses. The three isoforms were found to be distributed in a highly characteristic manner, suggesting that nerve cells have different requirements for the preservation of their intracellular calcium homeostasis.

Animals↗

PubMed1997

Ectopic expression of the calcium-binding protein parvalbumin in mouse liver endothelial cells.

To elucidate the physiological role of the Ca2+ binding protein parvalbumin, we have generated transgenic mice carrying the full-length complementary DNA (cDNA) of rat parvalbumin under the control of the heavy-metal inducible metallothionein IIA promoter. Immunohistochemical and biochemical methods have been used to detect the presence of ectopic parvalbumin expression in different tissues. Here we show the expression of parvalbumin in endothelial cells lining the liver sinusoids in situ and after isolation in vitro. The hemodynamic effects of endothelin 1, a peptide hormone mediating potent vasoconstriction via calcium signalling, were investigated in the mouse liver perfused in situ. Vasoconstriction, thought to be mediated by the Ito cell, was not affected in the transgenic animals, whereas microvascular exchange, probed with the multiple indicator dilution technique, was markedly decreased in normal mice but virtually not affected in the transgenic animals. This suggests that ectopically expressed parvalbumin is involved in the regulation of Ca2+ signals in the sinusoidal endothelial cells. This animal model could be of interest to those working on the physiology of liver circulation.

Animals↗

PubMed1997

HIV-I induced destruction of neocortical extracellular matrix components in AIDS victims.

Neurological dysfunction is not uncommon in patients suffering from acquired immunodeficiency syndrome (AIDS) and, when manifested, intimates involvement of the central nervous system. Here, the human immunodeficiency virus (HIV) infects preferentially microglial cells, which thereby release substances known to interfere with neuronal function. One class of agents set free in this manner are proteases; these degrade certain components within, and thereby undermine the integrity of, the extracellular matrix (ECM) compartment, which plays a vital role in cell-to-cell communication. We wished to ascertain whether the ECM compartment is indeed disrupted in the brains of AIDS victims. We examined the neocortical areas of 27 AIDS autopsy cases, including 9 with diagnosed HIV-encephalopathy (HIVE); 8 HIV-seronegative cases with various types of brain lesion, including viral infections, were also included in this study. HIV-antigens and DNA were identified by use of immunohistochemistry and in situ hybridization, and ECM components by lectin staining and immunohistochemistry. Of the 27 AIDS cases examined, each of the 9 with HIVE was completely devoid of labeled ECM components; 8 of the 18 without HIVE had incurred substantial losses, and only 2 manifested a normal complement of constituents within this compartment. With respect to stratal and topographic variations, layers II and III were less affected than layers V to VII, as was the frontal cortex relative to other areas. These findings confirmed our expectations of the brain's ECM undergoing degradation following HIV infection, and these changes may well underlie the neurological disturbances manifested in AIDS patients.

AIDS Dementia Complex↗

PubMed1997

Delayed decrease of calbindin immunoreactivity in the granule cell-mossy fibers after kainic acid-induced seizures.

Kainic acid (KA) administration induces an abnormal excitation and spontaneous recurrent seizures. Alterations of granule cell properties may be potential mechanisms. In this study, dynamic alterations of calbindin, a calcium binding protein particularly abundant in the granule cells, have been investigated immunocytochemically in the rat hippocampus after the KA-induced seizures. The calbindin immunoreactivity decreased slightly in the CA1/CA2 fields already after 1 and 3 days, and was lost partly or completely in the pyramidal layer after 10 days. From day 21, the calbindin immunoreactivity decreased in dendrites and soma of the granule cells and mossy fibers. The alterations remained at least to day 90, while no evident neuronal loss occurred in the granule cells. This may reflect a disturbance of calcium homostasis in the granule cells after seizures. The delayed decrease of calbindin has a time course similar to the occurrence of spontaneous recurrent seizures, suggesting a possible correlation between the two events.

Animals↗

PubMed1996

Change of calretinin expression in the human colon adenocarcinoma cell line HT29 after differentiation.

Calretinin is a Ca(2+)-binding protein of the EF-hand family which is expressed in colon adenocarcinomas and colon-derived tumor cell lines (e.g. WiDr), but is absent from normal human enterocytes. Its function has not as yet been elucidated, but some lines of evidence lead us to postulate its involvement in cell proliferation in these cells. In order to test whether calretinin is correlated with an undifferentiated, proliferating, or with a differentiated, state of cells, its expression was studied in the human colon adenocarcinoma clonal cell line HT29-18, which can be caused to differentiate into enterocyte-like cells by replacing glucose with galactose in the culture medium (glucose starvation differentiation). Treatment of HT29-18 cells with galactose led to a drop in the calretinin mRNA level and in protein expression as evidenced by immunocytochemical staining and Western blot analysis of cytosolic cell extracts. These results suggest that calretinin is present in HT29-18 cancer cells, mostly in those which are in the undifferentiated state. The possibility that calretinin is involved in maintaining the cells in an undifferentiated (cancerous) state is discussed.

Aminopeptidases↗

PubMed1996

Temporal and spatial appearance of the membrane cytoskeleton and perineuronal nets in the rat neocortex.

Parvalbumin-immunoreactive interneurons are surrounded by perineuronal nets, containing molecules of the extracellular matrix (e.g. tenascin-R). Furthermore, they seem to have a special cytoskeleton composed of, among others, ankyrinR and beta Rspectrin. In the present developmental study we showed that the intracellular markers parvalbumin, ankyrinR and beta Rspectrin as well as Vicia Villosa agglutinin, an extracellular marker for perineuronal nets, appeared in the second postnatal week. In the third postnatal week, ankyrinR and beta R spectrin were present in the parvalbumin-positive interneurons. Tenascin-R appeared in a similar topographic distribution as the intracellular markers. The adult pattern was established upon the end of the fourth postnatal week. Our results indicate that cytoskeletal maturity maybe a prerequisite for the organization of perineuronal nets of extracellular matrix.

Animals↗

PubMed1996

Rat brain oligodendrocytes do not interact selectively with axons expressing different calcium-binding proteins.

A single oligodendrocyte may endow ten to twenty vicinal axons with internodal segments, but its radial domain is neither exclusive of processes from other like cells nor are all nerve fibres within this zone myelinated. Whether oligodendrocytes are able to discriminate between axons on the basis of chemical or electrophysiological differences, or whether the tactic response is random, has yet to be established. In order to shed some light on this process, we investigated the ensheathment, by single oligodendrocytes, of axons distinguished on the basis of their calcium-binding protein complexion. Rat brain oligodendrocytes were visualized either with the Rip-antibody or by intracellular injection of Lucifer Yellow; subclasses of axons were immunolabelled with antibodies against one of the two calcium-binding proteins parvalbumin or calretinin. Individual oligodendrocytes did not exhibit exclusivity with respect to their preferment for axons containing calcium-binding proteins, associations with both non-immunoreactive, as well as with parvalbumin- or calretinin-positive ones, being encountered.

Animals↗

PubMed1996

Calretinin-immunoreactivity in organotypic cultures of the rat cerebral cortex: effects of serum deprivation.

Calretinin, a calcium-binding protein, is expressed in a specific set of interneurons in the adult rat cortex. Although its role in development is not known with any degree of certainty, evidence in support of a neuroprotective function has been forthcoming. To test this hypothesis, we submitted organotypic cultures (interphase technique) of 4- to 6-day-old rat brain slices to nutritive stress by serum deprivation for 1-3 weeks. Cultures were immunolabelled either with an antiserum against calretinin or with an antibody against MAP2 (the latter being used to assess neuronal cell number). In control (serum-enriched) cultures, the pattern of development of calretinin immunoreactivity mimicked that evinced in vivo with respect to layer- and cell-type specificity, but the maturation process was retarded by about 1 week. In the experimental group, cultures were incubated for 1 week in the presence of serum and then transferred to serum-free medium for an additional 2 weeks. Tissue was characterized by necrotic foci, a marked decrease in neuronal cell number and a further retardation in the course of development of the calretinin immunoreactivity pattern. The proportion of calretinin-immunoreactive cells to total number of viable neurons was 16% in serum-free cultures as against 9% in serum-enriched ones, suggesting that cells expressing the calcium-binding protein exhibit a greater tenacity for survival under conditions of nutritive stress, and thereby supporting the contention that calretinin acts in a neuroprotective capacity.

Animals↗

PubMed1996

Topographic variations in rat brain oligodendrocyte morphology elucidated by injection of Lucifer Yellow in fixed tissue slices.

Visualisation of oligodendrocytes by fluorochrome labelling in fresh tissue is a relatively recent innovation, but its widespread applicability in comparative analyses between different regions of the brain has been hampered by the limited survival time of excised preparations. We here applied the technique of impaling and injecting these cells with Lucifer Yellow in fixed tissue slices. Using confocal laser scanning microscopy, we reconstructed the three-dimensional forms of oligodendrocytes derived from the optic nerve, corpus callosum, cerebellum and spinal cord of young adult rats. Differences in shape and size of the cell body, in the number of internodal segments supplied by a single cell, as well in their spatial orientation, and in the thickness of the myelinated fibre, were observed between the four white matter tracts analysed.

Animals↗

PubMed1996

The proteoglycan DSD-1-PG occurs in perineuronal nets around parvalbumin-immunoreactive interneurons of the rat cerebral cortex.

Proteoglycans involved in the shaping of the developing brain are often preserved in the adult brain in more restricted locations. We have studied the fate of DSD-1-PG, a chondroitin sulfate proteoglycan containing the hybrid epitope DSD-1. DSD-1-PG exerts neurite outgrowth promoting activity and has been shown to occur in the developing brain during late brain development and into adulthood. In the adult rat brain, monoclonal and polyclonal antibodies against DSD-1-PG labelled only the circumference of a selected subpopulation of neurons. These nerve cells invariably expressed the calcium-binding protein parvalbumin. The label occupied the extracellular space in close vicinity to the cell body, surrounding axon terminals and glial end feet, but was absent from synaptic clefts. DSD-1-PG is thus shown to be an additional representative of the growing list of substances found in perineuronal locations in the adult mammalian brain.

Animals↗

PubMed1996

The calcium binding protein calretinin is a selective marker for malignant pleural mesotheliomas of the epithelial type.

In a series of 23 cases of mesothelioma of either the epithelial, sarcomatoid or the mixed type, the expression of three calcium-binding proteins (calretinin, parvalbumin and calbindin-D28k) was studied using immunohistochemical techniques on paraffin sections. The results show that calretinin is expressed in mesotheliomas of the epithelial type (papillary, adenomatous or solid) and by the epithelial component of the mixed tumours. The immunohistochemical reaction is specific and reproducible. The tissues of the pulmonary parenchyma and of the pleura are negative for calretinin except for the rare fibroblasts and some skeletal muscle fibres situated in the interstices of, or near the epithelial tumour mass. The sarcomatoid mesotheliomas and the sarcomatoid component of the mixed tumours do not express calretinin. Parvalbumin and calbindin-D28k are expressed neither in mesotheliomas nor in normal lung tissue. Primary adenocarcinomas of the lung are negative for all three calcium binding proteins cited. Thus, calretinin seems to represent a selective marker for mesotheliomas of the epithelial type and allows their differentiation from metastases of lung adenocarcinomas.

Adenocarcinoma↗

PubMed1996

Calretinin: a novel immunocytochemical marker for mesothelioma.

Immunohistochemistry is a powerful diagnostic adjunct in the differential diagnosis between malignant mesothelioma (especially of the epithelial type) and adenocarcinoma metastatic to the serous membranes. Most of the immunological probes commonly used, however, recognize antigens expressed by the epithelial malignancies and absent from mesothelial cells and mesotheliomas. Probes suitable for the positive identification of mesotheliomas are comparatively scarce and much less commonly used because of their reduced sensitivity and specificity, their unsuitability for staining routinely fixed and embedded tissues, or their lack of commercial availability. We now document that two different polyclonal antisera to calretinin consistently immunostain mesothelial cells and malignant mesotheliomas both in routinely fixed and embedded tissue sections and in cytological preparations of serous effusions. The diagnostic sensitivity of this novel immunocytochemical approach reached 100%, allowing immunostaining of all 44 mesotheliomas investigated, which included five biphasic and three sarcomatoid types. The specificity of calretinin immunoreactivity was checked against 294 adenocarcinomas of different origin (19 serosal metastases and 275 primary tumors potentially able to metastatize to serosal membranes) relevant for the discussion of the differential diagnosis with malignant mesothelioma: only 28 cases showed focal immunoreactivity for calretinin. We conclude that calretinin is a most useful marker for the positive identification of malignant mesotheliomas.

Adenocarcinoma↗