Biomedical subjects

J M Provis

Publications and source records attributed to J M Provis.

At least 37 records · Page 2Linked to original sources

PubMed1995

Development of microglial topography in human retina.

The development of microglial topography in wholemounts of human retina has been examined in the age range 10-25 weeks gestation (WG) using histochemistry and immunohistochemistry for CD45 and major histocompatibility complex class II antigens. Microglia were present in three planes corresponding to the developing nerve fibre layer/ganglion cell layer, the inner plexiform layer and the outer plexiform layer. Distribution patterns of cells through the retinal thickness and across the retinal surface area varied with gestational age. Microglia were elongated in superficial retina, large and ramified in the middle plane, and small, rounded and less ramified in deep retina. Intensely labeled, rounded profiles seen at the pars caeca of the ciliary processes, the retinal margin and at the optic disc may represent precursors of some retinal microglia. At 10 WG, the highest densities of microglia were present in middle and deep retina in the far periphery and at the retinal margin, with few superficial microglia evident centrally at the optic disc. At 14 WG, high densities of microglia were apparent superficially at the optic disc; microglia of middle and deep retina were distributed at more central locations although continuing to concentrate in the retinal periphery. Microglia appear to migrate into the developing human retina from two mains sources, the retinal margin and the optic disc, most likely originating from the blood vessels of the ciliary body and iris, and the retinal vasculature, respectively. The data suggest that the development of microglial topography occurs in two phases, an early phase occurring prior to vascularization, and a late phase associated with the development of the retinal vasculature.

Age Factors↗

PubMed1995

Human retinal microglia: expression of immune markers and relationship to the glia limitans.

The immunoreactivity, morphology and relationship to the glia limitans of microglia were investigated in flatmounts and sections of normal human retina, using immunogold histochemistry, electron microscopy (EM), and antibodies directed against CD45, major histocompatability complex class I (MHC-I), MHC-II, and human macrophage antigens. Immunoreactivity was evident for all antibodies tested, including MHC-I, which labeled both microglia and retinal vascular endothelium. Most consistent labeling was obtained using antibodies to CD45, MHC-II, and anti-human macrophage (S22) antigen. Immunoreactive cells were seen in the perivascular space (perivascular cells), where they were closely adherent to the vessel profile, and in the retinal parenchyma (microglia). Some parenchymal microglia were also vessel associated and by EM were seen to be closely related to the glia limitans (paravascular microglia). Paravascular microglia were shown by optical densitometry, to express higher levels of MHC antigens than neighboring, non-vessel associated, parenchymal microglia. In addition, paravascular microglia were macrophage (S22) antigen positive, while other parenchymal microglia did not express macrophage antigens. Quantitative data indicate that similar populations of microglia are immunoreactive to CD45, MHC-I, and MHC-II, while relatively few microglia (approximately 10%) are immunoreactive for human macrophage (S22) antigens, supporting previous suggestions that microglia are a heterogeneous population.

Adolescent↗

PubMed1995

Ontogeny and cellular expression of MHC and leucocyte antigens in human retina.

We have investigated the ontogeny of MHC class I, class II, CD45, and macrophage antigens in whole mounts of normal human fetal retina at 10-25 weeks gestation (WG) using monoclonal antibodies and immunogold histochemistry. MHC class I antigens were expressed on retinal vascular endothelial cells and provided a useful marker of vessel organization from 14-25 WG. Microglial cells expressed immunoreactivity to MHC class I, class II, and CD45 antigens from 10 WG (pre-vascularization) and macrophage S22 (Mac S22) antigen from 14 WG (post-vascularization), although none of the antigens tested were detected on neuronal or macroglial elements. Microglia expressing MHC, CD45, and macrophage antigens occurred in both ramified and rounded forms with no close correlation being observed between morphology and antigenicity. The numbers of immunoreactive cells labeled with each of the four markers increased steadily throughout gestation in all specimens studied. Equivalent numbers of microglia expressed MHC class I, class II, and CD45 antigens in retinae at similar gestational ages; however, our data indicate that microglia expressing Mac S22 antigen comprise approximately 40% or less of the population of MHC and CD45-immunoreactive cells during development. Topographical analyses suggest that MHC class I, class II, and CD45-positive microglia enter the retina from both the peripheral retinal margin and the optic disc from at least 10 WG; Mac S22-positive cells appear in association with the development of the retinal vasculature and enter the retina via the optic disc after 14 WG.

Biomarkers↗

PubMed1995

Immunohistochemical and topographic studies of dendritic cells and macrophages in human fetal cornea.

PURPOSE: To investigate the distribution and phenotype of major histocompatibility complex (MHC) class II-positive dendritic cells and macrophages in normal human fetal cornea in the age range 10 to 25 weeks gestation. METHODS: Peroxidase and gold immunohistochemistry were used to visualize MHC class II and macrophage antigen (S22) immunoreactive cells. Cell distributions were analyzed quantitatively, and topographic maps were produced. RESULTS: Immunoreactive cells, concentrated centrally, were present at 10 weeks gestation in the corneal epithelium and stroma. Average densities increased steadily up to 25 weeks gestation. Two morphologic forms of MHC class II and S22 immunoreactive cells were observed--large, dendritiform cells and small, rounded cells with short processes. Electron microscopy revealed that most MHC class II-positive cells were morphologically consistent with previous ultrastructural descriptions of corneal Langerhans cells. Immunoreactive cells were more numerous in immunogold-labeled specimens than in peroxidase-labeled specimens of similar ages. However, quantitative analysis of both techniques revealed that S22-positive cells comprised 30% to 50% of MHC class II-positive cells. CONCLUSIONS: This study provides a detailed description of heterogeneous populations of MHC class II and S22 immunoreactive cells in the human fetal cornea. In contrast to the adult cornea, which is typically devoid of MHC class II-positive cells, immunoreactive cells in the fetal cornea are concentrated centrally and increase in density up to at least 25 weeks gestation. These results indicate that reduction in Langerhans cell numbers to adult levels must occur after 25 weeks gestation. The presence of dendritic cells and macrophages in the fetal cornea has important implications for the understanding of corneal immunology.

Antibodies, Monoclonal↗

PubMed1994

Modulation of MHC class II expression in the absence of lymphocytic infiltrates in Alzheimer's retinae.

This study describes the expression of MHC class II antigens in retinal flat mounts from normal donors and patients with Alzheimer's disease (AD). We confirm previous observations of MHC class II immunoreactivity on microglia in normal retinae, while observing insignificant levels of reactivity on endothelial cells (EC). A significantly increased level of MHC class II expression was detected in AD retinae. This increased immunoreactivity was found to occur in the absence of lymphocytic infiltrates, suggesting that the pathogenesis of AD in the retina may be distinct from that reported to occur in some regions of the brain. MHC class II expression, measured using computerized optical densitometry, appeared to be increased principally as a result of induced MHC class II immunoreactivity on EC. Ramified microglia and perivascular macrophages, although hypertrophied, appeared to show unchanged levels of MHC class II expression. These findings are consistent with earlier suggestions that both aberrant MHC class II expression and suppressor activity of resident macrophages may restrict immune responses.

Adult↗

PubMed1994

Intermediate filament expression in human retinal macroglia. Histopathologic changes associated with age-related macular degeneration.

PURPOSE: Intermediate filament expression by retinal macroglia was studied in normal eyes, normal eyes of older subjects, and eyes of subjects with age-related macular degeneration (AMD), classified on the basis of histopathologic assessment of the retinal pigment epithelium (RPE), choroid, and donor age. METHODS: Adult human retinae (N = 43) were divided into three groups: normal (< 50 years of age, with normal RPE and choroid); normal eyes of older subjects (> 50 years with normal RPE and choroid); and eyes of subjects with AMD (> 50 years with histopathologic findings indicative of AMD), on the basis of histopathologic assessment of the RPE/choroid and donor age. Intermediate filament expression by retinal macroglia was studied in cryostat sections and in retinal flatmounts using avidin-biotin-peroxidase immunolabeling of antibodies to glial fibrillary acidic protein (GFAP) and vimentin. RESULTS. Analyses of immunohistochemically labeled retinal sections revealed similar distributions of vimentin reactivity in retinae of each group. Distributions of GFAP in normal and normal aged retinae were similar, but sections of AMD-affected retinae showed evidence of GFAP expression by Müller cells. In flatmounts, vimentin distribution was similar in the three groups, but GFAP labeling revealed hypertrophic astrocytes, which were absent from normal retinae, in 17% of aged retinae and 55% of AMD-affected retinae. Deeply penetrating, GFAP-positive processes were observed in 17%, 27%, and 55% of normal, normal aged, and AMD-affected retinae, respectively. CONCLUSIONS: Variation in GFAP and vimentin expression in retinal macroglia is affected by increasing age, and a distinctive variation of intermediate filament expression in retinal macroglia is associated with the pathogenesis of AMD.

Adolescent↗

PubMed1993

Human retinal microglia express phenotypic characteristics in common with dendritic antigen-presenting cells.

Neural tissue has been considered to be immunologically privileged and major histocompatibility complex (MHC) class II antigens not expressed in normal human brain grey matter and retina. In the present study we compare phenotypic characteristics of human retinal microglia and dendritic Langerhans cells, including their morphologies and distribution, MHC class II and CD45 antigen expression and nucleotidase reactivity. Levels of class II expression were measured using optical densitometry in combination with standard immunohistochemical techniques applied to retinal flatmounts. The results indicate that ramified retinal microglia have features in common with dendritic antigen presenting cells of cornea and conjunctivum, including the constitutive expression of MHC class II antigens.

Adult↗

PubMed1993

NADPH-diaphorase histochemistry reveals cone distributions in adult human retinae.

NADPH-diaphorase histochemistry was used to identify and analyse the topography of cones in adult human retinae (in the age range 44 to 68 years). Retinae were treated for NADPH-diaphorase reactivity, present in vascular walls, amacrine cells and cone and rod outer segments; a subpopulation of less intensely reactive profiles, morphologically resembling cones, possibly represents the blue cone population. Regularly spaced pairs of cones (one intensely labelled and one weakly labelled cone) were also apparent throughout the retina, and were most common along the horizontal meridian, particularly towards the periphery. The diameters and distributions of labelled cone outer segments were assessed using image analysis. Cone density in the adult retina ranged from 2000 per mm2 in the temporal periphery, to 82,000 to 120,000 per mm2 at the fovea centralis. Distribution patterns confirmed the presence of a cone streak, extending from the foveal region into nasal retina, but no evidence of superior-inferior asymmetry was detected.

Adult↗

PubMed1992

Evidence of photoreceptor migration during early foveal development: a quantitative analysis of human fetal retinae.

We have analyzed aspects of photoreceptor topography in wholemounts of human fetal retinae in the age range 13-24 weeks of gestation. Fetal retinae were stained with cresyl violet and the sizes and packing densities of rods and cones analyzed in the conventional manner. Cones and rods were present within a differentiating region, free of mitotic figures and approximately centered on the putative fovea, represented by the foveal cone mosaic. At 13 weeks of gestation the foveal cone mosaic was clearly differentiated, cone nuclei reaching a packing density of 14,200 per mm2; a small number of rods were present in the immediately adjacent region. The packing densities of both rods and cones in these regions gradually increased and the area of the foveal cone mosaic gradually decreased throughout the age range sampled, although individual variations were evident. By 24 weeks of gestation, cone density was approximately 38,000 per mm2 in the foveal cone mosaic. The maximum rod density observed was 59,200 per mm2 in the region surrounding the foveal cone mosaic in a specimen of 20-21 weeks of gestation. In all specimens, maximum cone density occurred within the foveal cone mosaic and gradually declined towards the periphery of the differentiating region; a pronounced inverse relationship between cone soma diameter and packing density was also observed. The evidence strongly suggests that both rods and cones migrate centripetally, that is towards the center of the developing fovea, from early in development, possibly from the time that they first differentiate. The implications of these findings for foveal development are discussed.

Cell Count↗

PubMed1991

NADPH-diaphorase reactivity in adult and developing cat retinae.

We have examined the distribution and size of nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase reactivity in adult and developing cat retinae. From late gestation E (embryonic day) 58 to adulthood, NADPH-diaphorase reactivity was detected in amacrine cells with somata located in the inner nuclear layer (INL) and ganglion cell layer (GCL) and in processes spreading in the middle strata of the inner plexiform layer (IPL). Reactivity was also present in small rounded profiles located in the outer plexiform layer (OPL) and thought to be cone pedicles. The number of NADPH-diaphorase reactive cells present in adult retinae was about 40,000, 75% of these somata were located in the GCL, the remainder in the INL. At birth, however, there was more than double this number of labelled somata (85,000), the total gradually declining to reach adult values by P (postnatal day) 25. This loss of NADPH-diaphorase reactive somata may be partly explained by natural cell death (apoptosis) or by loss of the active diaphorase from the cells. The density distributions of NADPH-diaphorase reactive cells in the INL and GCL of retinal wholemounts reached maxima in regions slightly inferior to the area centralis at all ages studied. The principal topographical difference between adult and developing retinae was that the density gradient of NADPH-diaphorase reactive cells was steeper in adults than at younger ages. During early development, the somal and dendritic field diameters of NADPH-diaphorase reactive cells at the area centralis were about the same size as those in the periphery; by adulthood, cells in the periphery were larger. The change in the somal diameter gradient apparently emerged because of a reduction in somal size of the centrally located cells. The change in the dendritic diameter gradient emerged because of a greater growth of peripheral cells as compared to central cells. We suggest that NADPH-diaphorase may have a role in the formation of synapses in the developing IPL.

Age Factors↗

PubMed1991

Antibodies to human leucocyte antigens indicate subpopulations of microglia in human retina.

Monoclonal antibodies to human leucocyte antigens, including anti-CD45 and anti-CD68, have been used to describe microglia in flatmounts of normal adult human retina for the first time. Anti-CD45 (the leucocyte common antigen) intensely labeled large numbers of cells in a regular distribution across the retina; anti-CD68 and anti-macrophage antibodies labeled fewer cells with distinctive morphologies, suggesting the presence of subpopulations of microglia in the human retina expressing leucocyte antigens.

Adult↗

PubMed1990

A distinctive soma size gradient among catecholaminergic neurones of human retinae.

We have examined the soma diameters and distribution of catecholaminergic (CA) cells in human retinae, by using an antibody to tyrosine hydroxylase (TH), the rate limiting enzyme in the production of catecholamines. TH-immunoreactivity was detected in two classes of cells (CA1 and CA2 cells). CA1 cells had relatively large somata (mean diameter 14 microns) located in either the inner nuclear layer (INL) or in the ganglion cell layer and extensive dendrites spreading into the other strata of the inner plexiform layer (IPL). CA2 cells had smaller, weakly labelled somata (mean diameter 9.6 microns) located principally in the inner regions of the INL and weakly labelled dendrites extending into the IPL. The mean density of CA2 cells in the far retinal periphery was approximately 38/mm2. The number of CA1 cells averaged approximately 15,600 per retina, with a mean density of 16/mm2. The density distribution of CA1 cells closely paralleled the distribution of ganglion cells, their density peaking at the foveal rim, with an area of relatively high density extending horizontally from the macula region toward the nasal margin (along the visual streak). A distinctive gradient was detected among the soma diameters of CA1 cells: they were largest in the mid-periphery, in a visual streak-like configuration around the optic disk. This gradient of soma size among CA cells closely corresponds to the density distribution of the rod photoreceptors in human retinae.

Animals↗

PubMed1990

Early differentiation of ganglion, amacrine, bipolar, and Muller cells in the developing fovea of human retina.

We examined the differentiation and maturation of neurons and glia of the inner nuclear layer (INL) and ganglion cell layer (GCL) in the retina of a human fetus of 15 weeks gestation. Serial, ultrathin sections were cut from a resin-embedded specimen from the posterior pole of the retina. The region of the putative fovea was defined by the absence of rod photoreceptors from the outer nuclear layer; only sections through the putative fovea were studied. Cell somata were classified on the basis of morphological criteria and, through the analysis of serial sections, morphological characteristics of the cell processes were established. In the inner plexiform layer (IPL), the types of synapses were analysed. The majority of cells in the INL and GCL were differentiated and could be identified. Ganglion cell somata were observed in the GCL and INL. Of 186 somata analysed in the INL, 66 were Muller cells, 21 amacrine cells, and 2 ganglion cells; a further 7 cells were classified as either amacrine or ganglion. Bipolar cells were thought to comprise the majority of the remaining 90 somata, but these could not be positively identified, as it was not possible to trace bipolar cell axons to their cell bodies deep in the INL. A detailed description of the morphological characteristics of the identified cells and their processes, and of the axonal processes of bipolar cells, is provided. Puncta adherentia and other simple intercellular junctions were commonly seen in the IPL and involved all cell types. Amacrine cell synapses and immature, monad bipolar cell synapses were common within the IPL. Dyad bipolar synapses were uncommon at this stage of development. A possible sequence of synaptogenesis in the IPL is discussed.

Cell Differentiation↗

PubMed1990

Angiogenesis in normal human retinal development: the involvement of astrocytes and macrophages.

Recent studies have suggested a role for mononuclear phagocytes series (MPS) cells in neovascularisation associated with retinal pathology and experimentally induced subretinal neovascularisation. The present study is concerned with the normal development of the human retinal vasculature. Morphological details are provided of developing vascular structures including the formation of tight junctions and canalisation of angioblast cords. The relationships of astrocytes and pericytes to developing structures and the presence of a perivascular collagenous matrix are described. Ultrastructural and histochemical analyses reveal an association between MPS cells and developing vascular structures. It is suggested that MPS cells may influence angiogenesis in normal retinal development, as well as in retinal pathology.

Astrocytes↗

PubMed1990

Autoantibodies to retinal astrocytes associated with age-related macular degeneration.

Sera from 128 patients with age-related macular degeneration (AMD) were examined and profiles of a variety of serum constituents, including immunoglobulins, alpha and beta globulins and autoantibodies, were tabulated. A similar series of tests were carried out on 20 control sera. The results indicate a higher incidence of serum abnormalities, particularly involving alpha-2 globulin, in patients with disturbance of pigmentation of the retinal pigment epithelium (RPE). The sera were further tested for the presence of autoantibodies with specificity for retinal tissue, and five major staining patterns were observed. Many sera produced patterns of labelling on human retina identical to that observed using labelled monoclonal anti-glial fibrillary acid protein (GFAP) antibodies, which are an established marker of retinal astrocytes. Although anti-retinal autoantibodies have been reported in association with a number of ocular pathologies, the observation of anti-astrocyte autoantibodies is new. Astrocytes are involved in the maintenance of the blood-retinal barrier (BRB) and also appear to be the facultative antigen-presenting cells of neural tissue. The present results indicate that the formation of anti-astrocyte autoantibodies may be an early feature of the pathogenesis of AMD.

Aged↗

PubMed1990

NADPH-diaphorase neurones of human retinae have a uniform topographical distribution.

We have examined the morphology and distribution of neurones that contain nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in human retinae. NADPH-diaphorase reactivity was observed in three different classes of amacrine cells (ND1, ND2, ND3 cells) and in the cone photoreceptors. ND1 cells had relatively large somata (mean, 12.3 microns) located in the inner nuclear layer (INL) and in the ganglion cell layer (GCL). Their dendrites were often strongly labeled and spread into either the middle or outer strata of the inner plexiform layer (IPL). The somata of ND2 cells were medium-sized (mean, 8.2 microns) and located in the INL and in the GCL; their dendrites were usually beaded and often spread in either the middle or outer strata of the IPL. ND3 cells had small, round somata (mean, 5.2 microns) located in either the INL or GCL, and were without labeled processes. The total number of NADPH-diaphorase cells (all classes) was estimated at 118,000, with a mean density of about 100/mm2. The most striking feature of NADPH-diaphorase cells in humans was that their distribution was relatively uniform across the retina, with no evidence of a peak in density at the foveal rim.

Cell Count↗

PubMed1989

Somatostatinergic neurones of the developing human and cat retinae.

We have examined somatostatin-immunoreactive (S-IR) neurones in developing retinae of the human and cat. At 14 and 16 weeks' gestation (G14 and G16) in the human, S-IR cells were only found close to the putative fovea centralis, but by 18 weeks' gestation (G18), they were located in all retinal regions. By adulthood, the majority of S-IR cells were restricted to inferior retina. In the developing cat retina, two classes of S-IR cells were recognized. S1-IR cells were similar in morphology and distribution to adult cells: they had small round somata which were only found in inferior retina and gave rise to beaded processes which traversed the inner plexiform layer (IPL) and nerve fibre layer (NFL). S2-IR cells had larger somata located in the ganglion cell layer (GCL) and the label was compartmentalized within their cytoplasm. Most S2-IR cells had lost immunoreactivity by P (postnatal day) 25 and may have been alpha-ganglion cells transiently expressing somatostatin in association with their retention of plasticity into postnatal life.

Animals↗

PubMed1988

Morphology of intraretinal new vessels in the PETH rat.

The mature stages of retinal dystrophy in PETH rats are characterised by loss of the photoreceptor layer and invasion of the retinal pigment epithelium by new capillaries derived from the retinal vessels. The new capillaries are fenestrated where they are adjacent to the basement membrane of the retinal pigment epithelium and are surrounded by cells of the disrupted pigment epithelium, which follow the course of the capillaries into the inner retina. Abnormal basement membrane deposits are common within the retinal pigment epithelium.

Animal Husbandry↗