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M G Stewart

Publications and source records attributed to M G Stewart.

At least 145 records · Page 8Linked to original sources
PubMed1987

Quantitative morphological effects of dark-rearing and light exposure on the synaptic connectivity of layer 4 in the rat visual cortex (area 17).

The quantitative effects of dark-rearing and light exposure on the ultrastructural characteristics of synapses and synaptic boutons in layer 4 of the rat visual cortex (area 17) have been investigated using stereological techniques. Two experimental groups (each containing 5 animals) were investigated-i) animals dark-reared upto weaning at 21 days post natum (21DPN) and then light exposed until 52DPN (Group 21/31), and ii) littermate animals totally dark-reared until 52DPN (Group 52dD). The results indicate a significantly higher mean density of synapses in the neuropil of layer 4 in group 21/31 (3.58 X 10(8).mm-3) compared with group 52dD (2.68 X 10(8).mm-3). Although the density per unit volume of synapses with identified asymmetrical synaptic membrane specialisations was not significantly different in group 21/31 than in group 52dD (but was significantly lower than animals reared normally), the density of synapses with identified symmetrical synaptic membrane specialisations was about 200% higher in group 21/31 versus group 52dD. However, significant differences were detected in the number of asymmetrical synapses established by single synaptic boutons in group 21/31 (1.21 +/- 0.11) compared with group 52dD (1.10 +/- 0.09). On the basis of the numbers of post-synaptic targets contacted by an individual synaptic bouton, a significantly higher density of synaptic boutons was found in group 21/31 (2.32 X 10(8).mm-3) compared with group 52dD (1.82 X 10(8).mm-3). Furthermore, planar quantitative data indicated significant intergroup differences in the ultrastructure of asymmetrical and symmetrical synaptic boutons. The results of this study provide evidence indicating marked structural alterations in the synaptic connectivity of layer 4 of the rat visual cortex following the light exposure of rats dark-reared upto weaning. Indeed visual deprivation severely affected the 'inhibitory' circuitry in the major thalamorecipient territory of the visual cortex.

Animal Husbandry↗
PubMed1987

Distribution of neurons and glia in the visual cortex (area 17) of the adult albino rat: a quantitative description.

The neuronal and glial cell composition of the rat visual cortex (area 17) has been determined quantitatively using stereological techniques. The volume numerical densities (number of cells per mm3 of cortex) of neurons and of the principal glial cell types (astroglia, oligodendroglia, and microglia) were calculated from tangential semithin resin sections spaced at regular intervals 50 micron apart throughout the entire depth of the visual cortex. From measurements of cortical and laminar thickness the separate volume numerical densities of neurons and glial cells were derived for each lamina in the cortex. In addition, the absolute numbers of cells in each lamina under 1 mm2 of cortical surface were calculated. The mean cortical volume numerical density of neurons was 60,020 +/- 3840/mm3 (mean +/- SEM; n = 8), and 49,040 +/- 2610/mm3 for the combined glial cell types. Astroglia, oligodendroglia, and microglia were present in a ratio of 6:3:1 respectively. It was determined from neuronal and glial somatic volume estimates that the somata of these cells occupied approximately 13.5% of unit cortical volume, with 81.3% of the unit volume being occupied by cortical neuropil. Using previously published reports that described the laminar composition of neurons in terms of the relative proportions of pyramidal and non-pyramidal cells, the laminar volume numerical densities for these neuronal categories have been derived. In addition, it has been estimated that under 1 mm2 of cortical surface there are 79,500 pyramidal and 7790 non-pyramidal neurons distributed throughout layers 1-6 of the rat visual cortex.

Animals↗
PubMed1986

GABA-immunoreactive neurons in the rat cerebellum: a light and electron microscope study.

An antibody raised against gamma - amino-butyric acid (GABA) coupled to bovine serum albumin with glutaraldehyde (Hodgson et al: J. Histochem. Cytochem. 33:229-239, '85) was used to localise immunocytochemically the presumptive GABAergic neuronal elements in the cerebellar cortex of the adult rat. employing the unlabelled antibody enzyme method with pre- and post-embedding immunocytochemical procedures, the following cellular structures were observed to be GABA-immunopositive in both the light and electron microscopes: the somata, dendrites, and axonal processes (including axon terminals) of stellate, basket, and Golgi neurons. In immunopositive neuronal somata and dendrites, the reaction product was found to be associated with all intracellular organelles and with the postsynaptic densities of synaptic junctions. Specific GABA-like immunoreactivity was also seen around outer mitochondrial membranes, microtubules, and neurofilaments, and coating synaptic vesicles in presynaptic axon terminals. In the pre-embedding procedure with dilutions of the antiserum between 1:1,000 and 1:2,000, the perikarya and dendrites of Purkinje cells were GABA-immunonegative, whereas at an antiserum dilution of 1:500 the somata of Purkinje cells were mildly GABA-immunoreactive. Purkinje cell axon terminals in the infra- and supraganglionic plexuses and in the deep cerebellar nuclei were always strongly immunopositive. Neuroglia were invariably GABA-immunonegative, as were the dendrites, axons (parallel fibres), and somata of granule cells. Mossy fibre and climbing fibre afferents were also immunonegative. The pattern of immunoreactivity obtained with this antiserum directed against the inhibitory neurotransmitter GABA was found to resemble closely the immunocytochemical distribution of GABA and of the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD) as reported previously in other immunocytochemical investigations (Oertel et al. and Wu et al: Cytochemical Methods in Neuroanatomy. New York: A. R. Liss, '82; Seguela et al: Neuroscience 16:865-874, '85; Mugnaini and Oertel: GABA and Neuropeptides in the CNS. Handbook of Chemical Neuroanatomy, Vol. 4, Part I. Amsterdam: Elsevier, '85.

4-Aminobutyrate Transaminase↗
PubMed1986

gamma-Aminobutyric acid immunoreactivity in mouse barrel field: a light microscopical study.

The barrel field of the mouse somatosensory cortex (SmI) was investigated immunocytochemically using an antiserum against the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). GABA-immunopositive cells and processes are grouped largely in the barrel side, whereas the barrel hollow is only weakly immunostained. The GABA-immunopositive cells have an ellipsoidal appearance similar to that of non-pyramidal class II barrel neurones described previously in Golgi impregnation studies of the mouse and rat barrel field.

Animals↗
PubMed1986

Decreased levels of an astrocytic marker, glial fibrillary acidic protein, in the visual cortex of dark-reared rats: measurement by enzyme-linked immunosorbent assay.

An antibody to glial fibrillary acidic protein (GFAP) (an immunocytochemical marker for astrocytes) has been used in an enzyme-linked immunosorbent assay (ELISA) to determine the amount of GFAP in three visual regions, the dorsal lateral geniculate nucleus (dLGN), the superior colliculus (SC) and the visual cortex (VC) (area 17) of dark-reared (D), normal (N) and light-exposed (L) rats. In all experiments GFAP was also measured in a control non-visual region, the motor cortex (MC) (area 4). No significant differences were found in GFAP in dLGN, SC or MC between D, L or N rats. However, in the visual cortex, the amount of GFAP in N rats was significantly greater than that in D rats (by 32%).

Animals↗
PubMed1986

The quantitative effects of dark-rearing and light exposure on the laminar composition and depth distribution of neurons and glia in the visual cortex (area 17) of the rat.

The effects of dark-rearing and light-exposure on the distribution of neurons and glial cells types in the rat visual cortex (area 17) have been investigated. Three groups of animals were studied: rats reared in the dark until weaning at 21 days post natum (21 DPN) and subsequently light-exposed for 31 days (Group 21/31); rats dark-reared until 52 DPN and then exposed to light for 3 days (Group 3 dL); and rats totally dark-reared until 52 DPN (Group 52 dD). Semithin sections tangential to the pial surface were obtained at sampling intervals 50 micron apart throughout the depth of the left visual cortex. The volume numerical densities of neurons, astroglia, oligodendroglia, and microglia, at each sampling strata in the cortex were calculated using stereological techniques. The laminer density and distribution of neurons was not significantly different between the three groups. In comparison with group 21/31 there was a marked reduction in the densities of astroglia, oligodendroglia, and microglia in lower layer 5 of groups 3 dL and 52 dD. Additionally, the density of microglia in thalamorecipeint layer 4 was greatly increased in group 3 dL compared with groups 21/31 and 52 dD. These results indicate specific alterations in the glial cell composition of the rat visual cortex following periods of dark-rearing and light-exposure. Furthermore, changes in the density of glial cells in layer 5 may reflect functional modifications in neurons projecting to the superior colliculus.

Animals↗
PubMed1986

GABA-immunoreactive neurons in the dorsal lateral geniculate nucleus of the rat: characterisation by combined Golgi-impregnation and immunocytochemistry.

Identified neurons in the dorsal lateral geniculate nucleus (dLGN) of the rat were studied using a Golgi/EM procedure in combination with post-embedding GABA immunocytochemistry. The results of this investigation identify clearly the GABA-immunoreactive nature of the two morphologically distinct neuronal populations found within the rat dLGN--thalamocortical relay cells are GABA-immunonegative whilst the putative 'interneurons' are GABA-immunopositive. This immunocytochemical evidence strongly suggests that 'interneurons' synthesize and use GABA to mediate their local inhibitory synaptic mechanisms within the dLGN of the rat.

Animals↗
PubMed1986

A quantitative investigation of the neuronal composition of the rat dorsal lateral geniculate nucleus using GABA-immunocytochemistry.

The proportion of neurons immunoreactive for gamma-aminobutyric acid (GABA), and their rostrocaudal distribution in the dorsal lateral geniculate nucleus of the rat, were determined quantitatively using post-embedding GABA-immunochemistry on semithin resin embedded coronal sections followed by stereological analysis. The mean total volume numerical density of neurons (total number of neurons per mm3) in the dLGN was 67,077 +/- 4412 mm-3 (mean +/- SEM; n = 5), comprising a mean volume numerical density for GABA-immunopositive neurons of 14,584 +/- 1324 mm-3, and a mean volume numerical density of GABA-immunonegative neurons of 52,493 +/- 3419 mm-3, GABA-immunopositive neurons constituted 21.7 +/- 0.5% of the total neuronal composition of the rat dorsal lateral geniculate nucleus. Although no rostrocaudal variation was detected in the total volume numerical density of neurons, the relative proportion of GABA-immunopositive neurons was significantly lower in the caudal segment (18.1 +/- 0.6%) compared with the middle (24.9 +/- 0.9%) and the rostral segments (22.1%). Furthermore, on the basis of somatic size distributions, GABA-immunonegative neurons were seen to be significantly smaller in the caudal segment than in the more anterior two segments. The somatic size of GABA-immunopositive neurons showed no rostrocaudal variation through the dorsal lateral geniculate nucleus. These data provide a morphological correlate for the structural and functional subdivision of the dorsal lateral geniculate nucleus described previously in electrophysiological and morphological studies.

Animals↗
PubMed1986

Development of gamma-aminobutyric acid immunoreactivity in chick hyperstriatum ventrale and cerebellum: light and electron microscopical observations.

The presence of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) in the avian hyperstriatum ventrale and dorsal cerebeller vermis was investigated immunocytochemically using a recently characterized antiserum raised against GABA. Tissue from domestic chicks aged from 19 days in ovo to 28 days posthatch was studied with both light and electron microscopy using pre-embedding immunocytochemistry. Basket, stellate and Golgi cells in the cerebellum, considered to be GABA-ergic, exhibited specific GABA-like immunolabelling in perikarya and in axonal and dendritic processes throughout the developmental period investigated. Purkinje cells also exhibited specific GABA-like immunoreactivity in both pre- and posthatch birds but the distribution and intensity of the immunolabelling varied with age and also in its location within the Purkinje neuron. In prehatch birds Purkinje perikarya exhibited heavy immunostaining which was substantially reduced posthatch, whereas the Purkinje primary dendrites remained immunopositive throughout the developmental period. A small population of cells in the medial hyperstriatum ventrale (mHV) were GABA-positive prehatch, but no immunopositive perikarya were evident in any posthatch samples. Small GABA-positive punctate profiles, representing boutons and transversely sectioned axons or dendrites, were present in the neuropil of all age groups studied. Possible reasons for these findings are discussed and it is suggested that the loss of perikaryal immunostaining, both in the cerebellar Purkinje cells and in those of the mHV, may be governed by maturational processes.

Animals↗
PubMed1985

GABA-immunoreactive neurons in the rat dorsal lateral geniculate nucleus: light microscopical observations.

The dorsal lateral geniculate nucleus (dLGN) of the rat was investigated immunocytochemically using an antiserum against the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). The appearance of GABA-immunopositive dendrites, dendritic appendages, and the size and shape of neuronal somata closely resembled the putative intrinsic neurons described previously in Golgi-impregnation studies of the rat dLGN.

Animals↗
PubMed1985

Effect of passive avoidance training on in vitro protein synthesis in forebrain slices of day-old chicks.

Slices from the forebrains of day-old chicks represent a highly active in vitro protein-synthesising system. The in vitro incorporation of L-[14C]leucine into protein of slices was estimated to be 2.5 mmol/mg protein/h. Incorporation was linear over 90 min of incubation and was suppressed by 92% by 1 mM cycloheximide. The highest incorporation was into microsomal and cell-soluble fractions. Under the electron microscope, slices appeared vacuolated near the cut surfaces, but well preserved internally (greater than 40 micron from the edge). Autoradiography showed that radioactivity was incorporated evenly across the slice with no decrease in label in the central part of the tissue. The rate of incorporation was only weakly dependent on leucine concentration in the medium (0.04-1 mM). Addition of a mixture of unlabelled amino acids (1 mM) produced a 20-50% inhibition of incorporation of radioactive L-leucine depending on the amino acids involved. In slices prepared from chicks 1 h after training on a one-trial passive avoidance paradigm, L-[14C]leucine incorporation was 23% higher (p less than 0.01) in the forebrain roof than in slices from control chicks. This figure is comparable to the one previously reported in vivo. Subcellular fractionation of incubated slices from the forebrain roof of trained and control birds revealed that the increased protein synthesis was due mainly to an elevated leucine incorporation into the soluble fraction.

Animals↗
PubMed1984

Hemispheric asymmetry of synapses in chick medial hyperstriatum ventrale following passive avoidance training: a stereological investigation.

A stereological analysis was made of synapses in the left and right hemispheres of chick medial hyperstriatum ventrale (MHV) 24h after passive avoidance training (PAL) and in water trained controls (W-control). The synaptic parameters examined were (D), the mean length of the postsynaptic thickening; (NV.syn), the number of synapses per unit volume of neuropil; (VV.syn), the volume density of the pre-synaptic bouton; (NV.ves), the number of synaptic vesicles per unit volume of neuropil and (ves.syn), the mean number of synaptic vesicles per pre-synaptic bouton. No significant differences exist in NV.syn between the left or right hemispheres of W-control and trained chicks, nor is NV.syn influenced by training. However, in W-control chicks D in the right MHV is significantly greater (12%) than in the left MHV and this difference disappears on training. There are no differences in VV.syn between left and right hemispheres of W-control chicks but following training VV.syn is 22.7% greater in the left MHV than in the right MHV. Training als influences the number of synaptic vesicles; in W-control chicks NV.ves in the right MHV is 12.25% greater than in the left MHV but following training these differences are reversed. When the data are expressed as numbers of vesicles per synapse (ves.syn), values for the left hemisphere of trained chicks exceed those in the right hemisphere by a staggering 61.38%. These results are discussed in the context of biochemical and electrophysiological studies which suggest that there is lateralization of the memory trace.

Animals↗
PubMed1981

Histochemical studies on the early proliferative lesion induced in the rat liver by aflatoxin.

Male inbred Fischer rats were fed a diet containing 5 p.p.m. aflatoxin for 1, 3, 4 1/2 and 6 weeks at which times groups were killed for histological and histochemical study. Aflatoxin produced a scattered individual cell necrosis of parenchymal cells by 1 week. At 3 weeks small basophilic proliferative foci were seen which increased in size and abundance to 6 weeks. These foci showed starvation-resistant glycogen, variable depletion of glucose-6-phosphatase, succinic dehydrogenase, aniline hydrogenase, membrane ATPase and acid phosphatase. At 6 weeks the foci showed the presence of gamma glutamyl transpeptidase and glucose-6-phosphate dehydrogenase. The basophilic foci were not preceded by other focal histological and histochemical change. The basophilic proliferative lesions are observed when an irreversible change has been induced in the liver. The role of such lesions in the histogenesis of hepatocellular carcinoma is discussed.

Aflatoxins↗
PubMedDate not supplied

Synapses in hippocampus occupy only 1-2% of cell membranes and are spaced less than half-micron apart: a quantitative ultrastructural analysis with discussion of physiological implications.

Relatively little information exists regarding the spatial structure of synaptic neuropil in the brain. The present electron microscopic study employs unbiased stereological techniques and Monte Carlo simulations to characterise quantitatively the spatial organisation of synaptic circuitry in the dentate gyrus of the hippocampus, an area of particular importance in mechanisms of learning and the subject of a number of experimental neurobiological models of synaptic plasticity such as long-term potentiation. Firstly, tissue shrinkage/expansion resulting from embedding was assessed by imaging 300-microm thick hippocampal slices in the course of the entire embedding protocol, giving a value of 94.3 +/- 1.1% for distance measures and 84.3 +/- 2.8% for volumetric measures. Secondly, numeric synaptic density, Nv, was estimated using the disector. Thirdly, accumulated area of post-synaptic densities (PSDs) per tissue volume, Sv, and the overall cell membrane area per tissue volume, Sv*, were assessed using unbiased stereological rules coupled with image analysis of single sections. Finally, the mean area of individual PSDs was derived as a ratio Sv/Nv giving: 0.0394 microm2 for axo-spinous PSDs (thus representing approximately 1.3% of total cell membranes) and 0.0769 microm2 for dendritic shaft PSDs (approximately 0.25% of total cell membranes). From these data, the mean nearest neighbour distance between synapses was estimated using Monte Carlo simulations of a random 3D arrangement of synapses constrained by PSD sizes (a truncated Poisson process), giving a value of 0.48-0.51 microm. The physiological importance of the morphometric data obtained is discussed in terms of assessing (i) the role of synaptic environment in modifying synaptic efficacy and (ii) the plausibility of cross talk between synapses in relation to extrasynaptic neurotransmitter diffusion and transient depletion of extracellular Ca2+.

Animals↗