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1.
Eur Rev Med Pharmacol Sci ; 19(3): 357-64, 2015.
Article in English | MEDLINE | ID: mdl-25720703

ABSTRACT

OBJECTIVE: Increase in ACL (anterior cruciate ligament) reconstructions has led to a higher prevalence of patients with postoperative symptoms which require investigation. We aimed to investigate the utility of magnetic resonance imaging (MRI) and computer tomography (CT) in determining tunnel size and graft obliquity after single bundle ACL reconstruction. PATIENTS AND METHODS: A retrospective comparison was made on 29 symptomatic knees after anatomic single bundle (trans AM) and transtibial ACL reconstructions which had both MRI and CT scans at an average of 1.3 years postoperatively (2 months-5.7 years). We compared CT and MRI (T2 sequence) tunnel size and graft obliquity estimates using Pearson correlation and t-test. We also compared MRI's of ACL reconstructed knees with hamstrings or patellar autografts, which were confirmed by operative protocol as either antero-medial (AM) technique (n=21) or trans-tibial (TT) technique (n=19). The surgeries were performed for an average of 6.29 (4-10) years for the TT group and 1.3 (0-3) years for the AM group, respectively. The graft inclination was measured relative to the tibial plateau using DICOM software. Statistical analysis used the mean value for each case and the data were processed using the non-parametric Kruskal-Wallis test to determine the difference in graft obliquity and tunnel placement. RESULTS: Tunnel size estimates correlate well between CT and MRI on axial scans: R2=0.795 and 0.630 for femur and tibia respectively. The position of the tunnels and graft obliquity were found to differ on MRI images in both coronal and sagittal planes. Coronal graft obliquity averaged 72.38° (ranging from 69° to 76°) using the AM technique and 75.47° (ranging from 72° to 78°) with TT technique. Sagittal graft inclination angle was 54.5 (51-58.5) and 63.68 (59-69.5) respectively. MRI proves to be the most useful imaging method in determining outcome after ACL reconstruction. However, for a better revision of the ACL reconstructions, CT can offer a clearer image of tunnels and bone stock. A more anatomical graft positioning increases obliquity in coronal and sagittal planes and, thus, becomes difficult to assess both tunnels in a single slice. CONCLUSIONS: The anatomic single bundle reconstruction technique has been found to more accurately reproduce the femoral footprint and the orientation of the graft compared to the TT technique where the appropriate tibial tunnel placement resulted in a more vertical graft.


Subject(s)
Anterior Cruciate Ligament Reconstruction/methods , Anterior Cruciate Ligament/diagnostic imaging , Anterior Cruciate Ligament/surgery , Graft Survival , Adult , Aged , Anterior Cruciate Ligament Reconstruction/standards , Female , Femur/diagnostic imaging , Femur/surgery , Humans , Imaging, Three-Dimensional/methods , Imaging, Three-Dimensional/standards , Knee Joint/diagnostic imaging , Knee Joint/surgery , Magnetic Resonance Imaging/methods , Magnetic Resonance Imaging/standards , Male , Middle Aged , Retrospective Studies , Tibia/diagnostic imaging , Tibia/surgery , Tomography, X-Ray Computed/methods , Tomography, X-Ray Computed/standards , Transplantation, Autologous/methods , Transplantation, Autologous/standards
2.
Eur Rev Med Pharmacol Sci ; 18(11): 1638-46, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24943975

ABSTRACT

OBJECTIVE: Besides than in the control of developmental events, axonal adhesive glycoproteins may be also involved in functions requiring fine organization and connectivity of the nervous tissue. We previously demonstrated morphological alterations and functional cerebellar deficits in transgenic mice (TAG/F3 mice) ectopically expressing the F3/Contactin axonal glycoprotein under the control of a selected regulatory region from the Transient Axonal Glycoprotein (TAG-1) gene. In the present study, the hippocampal function was explored by evaluating the ability of TAG/F3 mice to encode spatial and non-spatial relationships between discrete stimuli and to analyze an anxiety-related behavior. MATERIALS AND METHODS: To the first end, mice were placed in an "open-Field" containing five objects and, after three sessions of habituation (S2-S4), their reactivity to objects displacement (S5-S4) and object substitution (S7-S6) was examined.To the second end, mice were placed in the "elevated zero maze", a standard test to explore the anxiety-related behavior, in order to study, in transgenic mice, the effects of F3 misexpression on emotional reactivity by measuring the avoidance of the unsheltered open sectors. RESULTS: Statistical evaluations of reactivity to object novelty, TAG-F3 mice showed a lower DO exploration with respect to wild-type mice and, regarding DOs, TAG/F3 mice interacted less than wild-type mice, showing an impaired spatial change response. Furthermore, the number of HDIPS in transgenic TAG/F3 mice resulted significantly lower with respect to the controls (wild type). CONCLUSIONS: These results indicate that the coordinated expression of axonal adhesive glycoproteins may be relevant for the functional maturation of the hippocampus.


Subject(s)
Behavior, Animal/physiology , Contactin 1/physiology , Animals , Anxiety/genetics , Anxiety/psychology , Axons/physiology , Contactin 1/genetics , Female , Hippocampus/growth & development , Hippocampus/physiology , Male , Maze Learning/physiology , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Transgenic , Motor Activity/physiology , Pregnancy
3.
Eur Rev Med Pharmacol Sci ; 14(6): 507-12, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20712257

ABSTRACT

Rats receiving fluoride during the whole pregnancy up to the 9th day of lactation showed, when isolated at 10th day of life, a reduced rate of ultrasonic vocalizations (UV) in male pups (NaF 5.0 mg) and, in 90th days male rats, an increase of the Pre-Pulse Inhibition (PPI) with a reduction of the Peak response to the Startle stimulus given alone. Newborn rat reactivity could represent a useful and validated model in anxiety studies which could be moored with the Acoustic Startle Reflex (ASR) and PPI, appropriate models to study, in adulthood, particular neurological and psychiatric disorders showing deficits in attention and sensory-motor gating (Tourettes' syndrome, obsessive compulsive disorders, Huntington's disease and schizophrenia).


Subject(s)
Fetus/drug effects , Reflex, Acoustic/drug effects , Reflex, Startle/drug effects , Sodium Fluoride/toxicity , Vocalization, Animal/drug effects , Animals , Female , Male , Pregnancy , Rats , Rats, Wistar , Reflex, Acoustic/physiology , Reflex, Startle/physiology
4.
Neuroscience ; 161(4): 1057-66, 2009 Jul 21.
Article in English | MEDLINE | ID: mdl-19376199

ABSTRACT

Increased angiogenesis and an altered blood-brain barrier have been reported in the brain of dystrophin-deficient mdx mouse, an experimental model of Duchenne muscular dystrophy. To further elucidate the mechanisms underlying angiogenesis in Duchenne muscular dystrophy, in this study we evaluated whether nerve growth factor (NGF) and nerve growth factor receptors (NGFRs) are involved, then correlated NGF-NGFRs expression with vascular endothelial growth factor (VEGF) and its receptor-2 (VEGFR-2) content and matrix metalloproteinases-2 and -9 (MMP-2 and -9) activity, by confocal laser microscopy and immunohistochemistry. Results showed that neurons, astrocytes and ependymal cells were strongly labeled by NGF in mdx brain, expressing NGFRs on glial and endothelial cells. In controls, NGF faintly labeled neurons and astrocytes, whereas endothelial cells were negative for NGFRs. Immunogold electron microscopy demonstrated NGFR gold particles on endothelial cells in mdx brain, while in controls few particles were recognizable only on glial end feet. Western blotting and real time polymerase chain reaction (RT-PCR) demonstrated a higher expression of NGF and NGFR mRNA and protein in mdx brain as compared to controls, and increase of VEGF-VEGFR-2 and active MMP-2 and -9 content. Overall, these data suggest that in the brain of mdx mice, an upregulation of the NGF-NGFRs system might be involved directly, or indirectly through the activation of VEGF-VEGFR-2 and MMP-2 and -9, in the angiogenic response taking place in this pathological condition.


Subject(s)
Brain/metabolism , Muscular Dystrophies/metabolism , Nerve Growth Factor/metabolism , Receptor, trkA/metabolism , Receptors, Nerve Growth Factor/metabolism , Animals , Astrocytes/metabolism , Astrocytes/ultrastructure , Brain/ultrastructure , Ependyma/metabolism , Ependyma/ultrastructure , Female , Male , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mice , Mice, Inbred C57BL , Mice, Inbred mdx , Neuroglia/metabolism , Neuroglia/ultrastructure , Neurons/metabolism , Neurons/ultrastructure , RNA, Messenger/metabolism , Up-Regulation , Vascular Endothelial Growth Factor A/metabolism , Vascular Endothelial Growth Factor Receptor-2/metabolism
5.
Eur Rev Med Pharmacol Sci ; 12(3): 137-48, 2008.
Article in English | MEDLINE | ID: mdl-18700684

ABSTRACT

In the treatment of depression fluoxetine [a selective serotonine reuptake inhibitor (SSRIs)] is a widely used drug in humans. The selectivity, efficacy, side effects and simplicity of dosage contributed to fluoxetine's clinical acceptance. Several psychiatric disorders (many of them responsive to SSRIs) are present during pregnancy; up to 10% of pregnant women fulfill diagnostic criteria for major or minor depression with an even higher percentage developing postpartum depression. Therefore, significant numbers of women may be taking SSRIs while pregnant. Since fluoxetine's safe use during pregnancy is not yet established and experimental studies inconclusive, we performed the present research in order to investigate the neurobehavioral effects produced in rats by prenatal exposure to fluoxetine (5 and 10 mg/kg/sc from day 13 to 20 of gestation) on cognitive functions, emotional reactivity and sexual performance.


Subject(s)
Brain/drug effects , Fetus/drug effects , Fluoxetine/toxicity , Selective Serotonin Reuptake Inhibitors/toxicity , Abnormalities, Drug-Induced , Animals , Avoidance Learning/drug effects , Cognition/drug effects , Female , Male , Motor Activity/drug effects , Rats , Rats, Wistar , Receptors, Serotonin/drug effects , Sexual Behavior, Animal/drug effects , Vocalization, Animal/drug effects
6.
Neuroscience ; 149(3): 592-601, 2007 Nov 09.
Article in English | MEDLINE | ID: mdl-17916407

ABSTRACT

The aim of this study was to assess the effects of prenatal exposures to cannabinoids or carbon monoxide (CO) in an animal experimental model reproducing the environmental conditions in which a fetus develops whose mother, during pregnancy, ingests by smoking low doses of cannabinoids or CO. Particular attention was devoted to analyses of the long-term effects of the exposures at the level of the cerebellar cortex, where already during prenatal development the GABAergic neuronal systems may be modulated by both cannabinoids and CO. Three groups of rats were subjected to the following experimental conditions: exposure to cannabinoids by maternal treatment during pregnancy with the cannabinoid CB-1 receptor agonist WIN 55212-2 (WIN) (0.5 mg/kg/day, s.c.); exposure to CO by maternal exposure during pregnancy to CO (75 parts per million, by inhalation); and exposure to WIN+CO at the above doses and means of administration; a fourth group was used as control. The body weight of dams, length of pregnancy, litter size at birth, body weight and postnatal mortality of pups were monitored in order to evaluate possible effects of the exposures on reproduction and on prenatal and postnatal development. In the different groups, the long-term effects of the exposures were studied in adult rats (120-150 days) by light microscopy analyses of the structure of the cerebellar cortex and of the distribution in the cortex of markers of GABAergic neurons, such as GAD and GABA itself. Results. Exposures to WIN or CO did not affect reproduction or prenatal/postnatal development. Moreover, the exposed rats showed no structural alterations of the cerebellar cortex and displayed qualitative distribution patterns of GAD and GABA immunoreactivities similar to those of the controls. However, quantitative analyses indicated significant changes of both of these immunoreactivities: in comparison with the controls, they were significantly increased in WIN-exposed rats and reduced in CO-exposed rats, but not significantly different in WIN+CO-exposed rats. The changes were detected in the molecular and Purkinje neuron layers, but not in the granular layer. Prenatal exposures of rats to WIN or CO, at doses that do not affect reproduction, general processes of development and histomorphogenesis of the cerebellar cortex, cause significant changes of GAD and GABA immunoreactivities in some GABAergic neuronal systems of the adult rat cerebellar cortex, indicating selective up-regulation of GABA-mediated neurotransmission as a long-term consequence of chronic prenatal exposures to cannabinoids or CO. Because the changes consist of overexpression or, vice versa, underexpression of these immunoreactivities, functional alterations of opposite types in the GABAergic systems of the cerebellum following exposure to WIN or CO can be postulated, in agreement with the results of behavioral and clinical studies. No changes in immunoreactivities were detected after prenatal exposure to WIN and CO in association.


Subject(s)
Benzoxazines/pharmacology , Carbon Monoxide/pharmacology , Cerebellar Cortex/physiology , Morpholines/pharmacology , Naphthalenes/pharmacology , Neurons/physiology , Receptor, Cannabinoid, CB1/agonists , gamma-Aminobutyric Acid/physiology , Animals , Atmosphere Exposure Chambers , Birth Weight/drug effects , Body Weight/drug effects , Cerebellar Cortex/cytology , Cerebellar Cortex/drug effects , Female , Glutamate Decarboxylase/metabolism , Pregnancy , Prenatal Exposure Delayed Effects , Presynaptic Terminals/drug effects , Rats , Rats, Wistar , Reproduction/drug effects , gamma-Aminobutyric Acid/metabolism
7.
Eur J Histochem ; 51 Suppl 1: 59-64, 2007.
Article in English | MEDLINE | ID: mdl-17703595

ABSTRACT

The granular layer of the cerebellar cortex is composed of two groups of neurons, the granule neurons and the so-called large neurons. These latter include the neuron of Golgi and a number of other, lesser known neuron types, generically indicated as non-traditional large neurons. In the last few years, owing to the development of improved histological and histochemical techniques for studying morphological and chemical features of these neurons, some non-traditional large neurons have been morphologically well characterized, namely the neuron of Lugaro, the synarmotic neuron, the unipolar brush neuron, the candelabrum neuron and the perivascular neuron. Some types of non-traditional large neurons may be involved in the modulation of cortical intrinsic circuits, establishing connections among neurons distributed throughout the cortex, and acting as inhibitory interneurons (i.e., Lugaro and candelabrum neurons) or as excitatory ones (i.e., unipolar brush neuron). On the other hand, the synarmotic neuron could be involved in extrinsic circuits, projecting to deep cerebellar nuclei or to another cortex regions in the same or in a different folium. Finally, the perivascular neuron may intervene in the intrinsic regulation of the cortex microcirculation.


Subject(s)
Cerebellar Cortex/ultrastructure , Neurons/ultrastructure , Animals , Cell Communication , Humans
8.
Neuroscience ; 135(3): 897-905, 2005.
Article in English | MEDLINE | ID: mdl-16112480

ABSTRACT

Glutamic acid decarboxylase and GABA immunoreactivities were qualitatively and quantitatively evaluated in the cerebellar cortex of adult rats prenatally exposed to a low concentration of carbon monoxide (75 parts per million). Carbon monoxide-exposed and control rats were perfused with modified Bouin's fluid and their cerebella were embedded in paraffin. Sections from the vermis of each cerebellum were stained with Toluidine Blue or assayed with anti-glutamic acid decarboxylase 65/67 or with anti-GABA antisera. In the Toluidine Blue-stained sections, no differences were observed in the microscopic structure of the cerebellar cortex between carbon monoxide-exposed rats and controls. The distribution patterns of glutamic acid decarboxylase and GABA immunoreactivities in the cerebellar cortex of the treated animals were qualitatively comparable to those of the controls, and in accordance with previous descriptions of glutamic acid decarboxylase and GABA immunoreactivities in the rat cerebellar cortex. However, quantitative analyses demonstrated a significant reduction of immunoreactivities to both substances in the exposed rats in comparison with the controls. The reduction regarded: in the molecular layer, the number of glutamic acid decarboxylase/GABA-immunoreactive neuronal bodies and of axon terminals and the area they covered; in the Purkinje neuron layer, the number and the area covered by glutamic acid decarboxylase/GABA immunoreactive axon terminals. The differences detected in the prenatally exposed adult rats could be due to carbon monoxide-induced impairment of the differentiation of cerebellar GABA synthesizing neurons. A consequently diminished synthesis of GABA might account for some behavioral disorders detected in adult rats submitted to the same experimental procedure.


Subject(s)
Carbon Monoxide/toxicity , Cerebellar Cortex/metabolism , Glutamate Decarboxylase/metabolism , Prenatal Exposure Delayed Effects , gamma-Aminobutyric Acid/metabolism , Animals , Carboxyhemoglobin/metabolism , Cerebellar Cortex/enzymology , Coloring Agents , Cytoplasmic Granules/enzymology , Cytoplasmic Granules/metabolism , Female , Immunohistochemistry , Male , Pregnancy , Purkinje Cells/enzymology , Purkinje Cells/metabolism , Rats , Rats, Wistar , Smoking/metabolism , Tolonium Chloride
9.
Anat Embryol (Berl) ; 208(1): 55-64, 2004 Apr.
Article in English | MEDLINE | ID: mdl-15014985

ABSTRACT

'Non-traditional' large neurons of the granular layer of the cerebellar cortex include all its large neuronal types, except the Golgi neuron, which is instead one of the five 'classic' types of corticocerebellar neurons. The morphological, chemical and functional characteristics of the 'non-traditional' large neurons have not been entirely ascertained. The aim of this study was to ascertain whether morphological evidence can be provided of GABA synthesis within the 'non-traditional' large neurons of the human cerebellar cortex by means of immunocytochemistry for glutamic acid decarboxylase (GAD). Fragments of postmortem cerebellar cortex of various lobules from the hemispheres and vermis were studied. Immunoreactions revealed large neurons distributed throughout the granular layer in all lobules examined. They were discriminated by analyzing the morphological features of their bodies and processes and were identified as Golgi neurons and as some 'non-traditional' types, such as the candelabrum, Lugaro and synarmotic neurons. In addition, immunoreactive large neurons, with their bodies and processes closely adjacent to microvessels, were observed throughout the layer: these perivascular neurons could represent a new type of 'non-traditional' neuron of the cerebellar cortex. This study supplies the first indication that in the human cerebellar cortex some types of 'non-traditional' large neurons are GAD-immunoreactive, in addition to those neurons already known to be GABAergic (i.e., stellate, basket, Purkinje and Golgi neurons). These morphological data further point out possible functional roles for GABA as a neurotransmitter/neuromodulator in intrinsic, associative and projective circuits of the cerebellar cortex.


Subject(s)
Cerebellar Cortex/enzymology , Glutamate Decarboxylase/metabolism , Neurons/enzymology , Adult , Cadaver , Cell Size , Cerebellar Cortex/cytology , Humans , Immunohistochemistry , Isoenzymes/metabolism , Middle Aged , Neurons/cytology , Tissue Distribution
10.
Eur J Histochem ; 47(3): 253-6, 2003.
Article in English | MEDLINE | ID: mdl-14514417

ABSTRACT

A number of immunocytochemical studies have indicated the presence of cholinergic neurons in the cerebral cortex of various species of mammals. Whether such cholinergic neurons in the human cerebral cortex are exclusively of subcortical origin is still debated. In this immunocytochemical study, the existence of cortical cholinergic neurons was investigated on surgical samples of human parietal association neocortex using a highly specific monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine biosynthesising enzyme. ChAT immunoreactivity was detected in a subpopulation of neurons located in layers II and III. These were small or medium-sized pyramidal neurons which showed cytoplasmic immunoreactivity in the perikarya and processes, often in close association to blood microvessels. This study, providing demonstration of ChAT neurons in the human parietal neocortex, strongly supports the existence of intrinsic cholinergic innervation of the human neocortex. It is likely that these neurons contribute to the cholinergic innervation of the intracortical microvessels.


Subject(s)
Choline O-Acetyltransferase/metabolism , Neocortex/cytology , Neocortex/enzymology , Neurons/enzymology , Aged , Antibodies, Monoclonal/immunology , Choline O-Acetyltransferase/immunology , Humans , Middle Aged
11.
Neuroscience ; 115(1): 145-52, 2002.
Article in English | MEDLINE | ID: mdl-12401329

ABSTRACT

Caveolae are microinvaginations of the cell plasma membrane involved in cell transport and metabolism as well as in signal transduction; these functions depend on the presence of integral proteins named caveolins in the caveolar frame. In the brain, various caveolin subtypes have been detected in vivo by immunocytochemistry: caveolin-1 and -2 were found in rat brain microvessels, caveolin-3 was revealed in astrocytes. The aim of this study was to identify the site(s) of cellular expression of caveolin-1 in the microvessels of the human cerebral cortex by immunofluorescence confocal microscopy and immunogold electron microscopy. Since in the barrier-provided brain microvessels tight relations occur between the endothelium-pericyte layer and the surrounding vascular astrocytes, double immunostaining with caveolin-1 and the astroglia marker, glial fibrillary acidic protein, was also carried out. Immunocytochemistry by confocal microscopy revealed that caveolin-1 is expressed by endothelial cells and pericytes in all the cortex microvessels; caveolin-1 is also expressed by cells located in the neuropil around the microvessels and identified as astrocytes. Study of the cortex microvessels carried out by immunoelectron microscopy confirmed that in the vascular wall caveolin-1 is expressed by endothelial cells, pericytes, and vascular astrocytes, and revealed the association of caveolin-1 with the cell caveolar compartment. The demonstration of caveolin-1 in the cells of the brain microvessels suggests that caveolin-1 may be involved in blood-brain barrier functioning, and also supports co-ordinated activities between these cells.


Subject(s)
Caveolins/biosynthesis , Cerebral Cortex/blood supply , Cerebral Cortex/metabolism , Blood-Brain Barrier/physiology , Caveolin 1 , Caveolins/analysis , Cerebral Cortex/chemistry , Humans , Microcirculation/chemistry , Microcirculation/metabolism , Middle Aged
13.
J Submicrosc Cytol Pathol ; 33(3): 261-73, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11846095

ABSTRACT

The developmental effects of lead acetate were studied in the chick embryo metanephros, the third renal rudiment that acquires morphological characteristics of functioning kidney already during the prenatal life. Lead exposure was obtained by applying a lead acetate solution on the chick embryo chorioallantoic membrane at the days 9, 10 and 11 of incubation. Quantitative evaluation of the lead concentration assessed by furnace atomic absorption spectrophotometry at the days 14 and 21 of incubation demonstrated metal presence both in the chorioallantoic membrane (CAM) and in metanephros (MN). The lead concentration was higher in CAM than in MN; the metal amount was similar in the CAM of 14 and 21 day embryos, but significantly higher in the 14day embryo MN than in the 21 day embryo MN. Morphological observations on metanephros tissue of control and lead-treated embryos were performed under light, electron transmission and electron scanning microscopes. Peculiar attention was devoted to the expression of the junctional protein connexin 43, the major component of the gap junctions in the renal cells. The results indicated that lead treatment does not intervene in the general differentiation of the metanephric nephrons. The lead is reabsorbed by the proximal tubule cells that are engulfed by endocytotic vacuoles and metal deposits and show long term degenerative changes. Expression of Cx43 protein and ultrastructure of gap junctions between proximal tubule cells appeared to be unchanged. The morphological aspects of the MN corpuscles and tubules agree with the suggestion of a lead cytotoxic effect but do not corroborate, at least in this experimental model, the view of primary damage exerted by lead on the gap junctions of the renal epithelial cells.


Subject(s)
Chick Embryo/drug effects , Kidney/drug effects , Lead/toxicity , Organometallic Compounds/toxicity , Abnormalities, Drug-Induced , Allantois/chemistry , Allantois/drug effects , Animals , Chick Embryo/abnormalities , Chorion/chemistry , Chorion/drug effects , Connexin 43/analysis , Embryonic and Fetal Development , Gap Junctions/drug effects , Gap Junctions/ultrastructure , Kidney/abnormalities , Lead/analysis , Lead/pharmacokinetics , Organometallic Compounds/pharmacokinetics
14.
Histochem J ; 33(9-10): 537-43, 2001.
Article in English | MEDLINE | ID: mdl-12005025

ABSTRACT

The distribution of gamma-aminobutyric acid (GABA) in surgical samples of human cerebellar cortex was studied by light and electron microscope immunocytochemistry using a polyclonal antibody generated in rabbit against GABA coupled to bovine serum albumin with glutaraldehyde. Observations by light microscopy revealed immunostained neuronal bodies and processes as well as axon terminals in all layers of the cerebellar cortex. Perikarya of stellate, basket and Golgi neurons showed evident GABA immunoreactivity. In contrast, perikarya of Purkinje neurons appeared to be negative or weakly positive. Immunoreactive tracts of longitudinally- or obliquely-sectioned neuronal processes and punctate elements, corresponding to axon terminals or cross-sectioned neuronal processes, showed a layer-specific pattern of distribution and were seen on the surface of neuronal bodies, in the neuropil and at microvessel walls. Electron microscope observations mainly focussed on the analysis of GABA-labelled axon terminals and of their relationships with neurons and microvessels. GABA-labelled terminals contained gold particles associated with pleomorphic vesicles and mitochondria and established symmetric synapses with neuronal bodies and dendrites in all cortex layers. GABA-labelled terminals associated with capillaries were seen to contact the perivascular glial processes, basal lamina and endothelial cells and to establish synapses with subendothelial unlabelled axons.


Subject(s)
Cerebellar Cortex/chemistry , gamma-Aminobutyric Acid/analysis , Aged , Animals , Cerebellar Cortex/ultrastructure , Female , Humans , Immunohistochemistry , Male , Middle Aged , Neurons/metabolism , Neurons/ultrastructure
15.
Histochem J ; 32(9): 557-64, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11127977

ABSTRACT

The distribution of gamma-aminobutyric acid (GABA) in the human cerebellar cortex was studied using immunohistochemistry for glutamic acid decarboxylase (GAD), the enzyme that catalyses GABA synthesis. Observations by light microscopy revealed, in all layers of the cerebellar cortex, strong, punctate positivity for GAD, related to putative GABAergic nerve terminals, as well as a diffuse cytoplasmic immunoreactivity within neuronal cell bodies. GAD-positive nerve terminals were found in close relationship with the walls of the cerebellar cortex microvessels. Observations by electron microscopy revealed positive nerve terminals in contact with the astrocyte perivascular sheath of capillaries. GAD immunoreactivity was also detected within astroglial perivascular endfeet and endothelial cells. The findings provide further insights into the GABAergic synapses of the circuitry of the human cerebellar cortex. The detection of 'vascular' GAD immunoreactivities suggests that GABAergic mechanisms may regulate cerebellar microvessel function.


Subject(s)
Cerebellar Cortex/enzymology , Glutamate Decarboxylase/analysis , Isoenzymes/analysis , Neurons/enzymology , Aged , Capillaries/enzymology , Cerebellar Cortex/blood supply , Cerebellar Cortex/cytology , Cerebellar Cortex/pathology , Glutamate Decarboxylase/immunology , Humans , Isoenzymes/immunology , Middle Aged , Purkinje Cells/cytology , Purkinje Cells/enzymology
16.
Histochem Cell Biol ; 114(3): 191-5, 2000 Sep.
Article in English | MEDLINE | ID: mdl-11083461

ABSTRACT

The distribution of glutamic acid decarboxylase (GAD), the gamma-aminobutyric acid (GABA) synthesizing enzyme, was examined in the postmortem human cerebellar cortex by immunocytochemistry. The results, obtained on nervous tissues taken at autopsy and fixed within 24-36 h after death, enabled the authors to precisely reveal the topographical distribution of GAD-containing neurons and axon terminals in the human cerebellar cortex. Labeled neurons, corresponding to different neuronal cytotypes of the cerebellar cortex, showed a diffuse cytoplasmic immunoreactivity in both bodies and processes. Labeled axon terminals appeared as immunoreactive puncta. The use of immunocytochemistry in the detection of GAD in the postmortem human brain greatly increases the possibility of carrying out morphological studies on the GABAergic system, both in normal and in pathological conditions.


Subject(s)
Cerebellar Cortex/enzymology , Glutamate Decarboxylase/analysis , Neurons/enzymology , Purkinje Cells/enzymology , Adult , Cadaver , Cerebellar Cortex/cytology , Humans , Immunohistochemistry/methods , Middle Aged , Neurons/cytology , Purkinje Cells/cytology
17.
Brain Res Dev Brain Res ; 123(1): 95-101, 2000 Sep 30.
Article in English | MEDLINE | ID: mdl-11020555

ABSTRACT

The blood-brain barrier (BBB) glucose transporter, GLUT1, was detected by immunogold electron microscopy on the microvascular compartment of the human foetus telencephalon at the 12th and 18th weeks of gestation. By computerized morphometry, the cellular and subcellular localization of the immunosignal for GLUT1 was quantitatively evaluated. The study showed that the glucose transporter is strongly expressed by endothelial cells while a very low signal is detected on vascular pericytes. The GLUT1 antigenic sites are preferentially associated to the ablumenal and junctional plasma membranes of the endothelial cells and tend to increase significantly with age. A parallel study carried out by the endogenous serum protein albumin demonstrated that already at the 12th week the endothelial routes are hindered to the protein as happens at the blood-endothelium interface of mature brain. The results demonstrate that in the human foetus the brain microvessels express BBB-specific functional activities early.


Subject(s)
Blood-Brain Barrier/physiology , Brain Chemistry/physiology , Brain/anatomy & histology , Monosaccharide Transport Proteins/metabolism , Serum Albumin/metabolism , Female , Glucose Transporter Type 1 , Humans , Image Interpretation, Computer-Assisted , Immunohistochemistry , Microscopy, Immunoelectron , Pregnancy , Subcellular Fractions/drug effects , Subcellular Fractions/metabolism
18.
Inflamm Res ; 49(8): 418-23, 2000 Aug.
Article in English | MEDLINE | ID: mdl-11028759

ABSTRACT

OBJECTIVE AND DESIGN: Angiogenesis plays an important role in the pathogenesis of rheumatoid arthritis (RA) and correlates with clinical score, synovial hyperplasia and infiltration of inflammatory cells. Many of the available treatments for RA have been shown to possess some degree of anti-angiogenic activity. Here, we studied the effect of cyclosporine, which exerts anti-angiogenic activity in vitro and in vivo [1] on angiogenesis induced in vivo in the chick embryo chorioallantoic membrane (CAM) by synovial RA and osteoarthritis (OA) tissues. MATERIAL AND METHODS: Wet synovial biopsies from 10 RA and 6 OA patients were treated with vehicle alone or with cyclosporine and implanted on the CAM at day 8 of incubation. On day 12, CAM tissues were assessed for the extent of angiogenesis and mononuclear cell infiltration. RESULTS: Cyclosporine inhibited angiogenesis and reduced the number of mononuclear cells in the CAM extracellular matrix only in RA implants. CONCLUSIONS: These data provide further evidence for a central role of new-formed blood vessels in RA. Moreover, cyclosporine on account of both its immunosuppressive and its anti-angiogenic activity can be proposed for the treatment of RA.


Subject(s)
Allantois/blood supply , Arthritis, Rheumatoid/physiopathology , Chorion/blood supply , Cyclosporine/therapeutic use , Neovascularization, Pathologic/prevention & control , Osteoarthritis/physiopathology , Animals , Chick Embryo , Humans , Synovial Membrane/transplantation , Time Factors
19.
Eur J Histochem ; 44(2): 165-9, 2000.
Article in English | MEDLINE | ID: mdl-10968364

ABSTRACT

The distribution of cholinergic nerve fibres associated with the microvasculature of the human parietal cerebral cortex was investigated by immunocytochemistry, employing monoclonal antibodies against choline acetyl-transferase, the acetylcholine-synthesizing enzyme. The results revealed strongly immunoreactive nerve fibres in the tunica adventitia of arterioles penetrating the superficial cortical layers from the pial vasculature. Networks of stained nerve fibres were seen within the tunica muscularis of the radially directed arterioles that cross the intermediate and deep cortical laminae, and of their transverse and recurrent branches. Tiny positive nerve fibres were also seen around the cortex capillaries, some reaching the endothelial cells. The morphological data support the involvement of acetylcholine in microvasculature local regulation, possibly with a differentiated role in the arterioles and capillaries.


Subject(s)
Arterioles/innervation , Cerebral Arteries/innervation , Choline O-Acetyltransferase/analysis , Nerve Fibers/enzymology , Parietal Lobe/blood supply , Antibodies, Monoclonal/immunology , Arterioles/pathology , Capillaries/innervation , Cerebral Arteries/pathology , Choline O-Acetyltransferase/immunology , Humans , Immunoenzyme Techniques , Nerve Fibers/pathology , Parietal Lobe/pathology
20.
Neuroreport ; 7(10): 1577-81, 1996 Jul 08.
Article in English | MEDLINE | ID: mdl-8904758

ABSTRACT

Vasoactive intestinal polypeptide-like immunoreactive (VIP-LIR) astrocytes were found in the subcortical white matter of the human forebrain parietal lobe. Astrocytes expressing VIP-LIR represented a minority (0.97%) of the GFAP-stained astrocyte population in the white matter. The close anatomical relationship between the VIP-LIR astrocyte bodies and processes and the brain vasculature strongly suggests that they may play a role in the local control of blood flow and of the barrier properties of the vessel walls.


Subject(s)
Astrocytes/metabolism , Brain/metabolism , Vasoactive Intestinal Peptide/metabolism , Humans , Image Processing, Computer-Assisted , Immunohistochemistry
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