Search PubMed⌕ Search

Biomedical subjects

M G Stewart

Publications and source records attributed to M G Stewart.

At least 127 records · Page 7Linked to original sources

Quantitation of synaptic, neuronal and glial development in the intermediate and medial hyperstriatum ventrale (IMHV) of the chick Gallus domesticus, pre- and post-hatch.

A quantitative analysis was made of the development of synapses, neurons and glia in both left and right intermediate and medial hyperstriatum ventrale (IMVH) of the forebrain of the domestic chick, Gallus domesticus from 16 days in ovo to 9 days post-hatch. There was a marked increase in total synapse numerical density (NVsyn), from 10 synapses per 100 microns3 at 16 days in ovo to 50 synapses per 100 microns3 at 9 days post-hatch; no significant left/right hemispheric differences were evident but there were differences between the development profiles for synapses with asymmetric as compared to symmetrical synaptic junctions. In contrast to synaptic development, the number of neurons per unit volume, NVneu, decreased by approximately 50% from the value at 16 days in ovo to that at day 0 (hatching). The neuronal population density remained unchanged to 3 days post-hatch and then, in 9-day-old birds, declined to almost a quarter of the original population size; no significant left/right hemisphere differences were evident. Glial cells declined in number from 16 days in ovo to 19 days in ovo, and then gradually increased to 9 days post-hatch. When the synapse to neuron ratio was examined, a trend was observed of a gradual increase with age, but no hemispheric differences were present. It is concluded that these changes are major events which must be considered in experiments aimed at determining the effects of behavioural, and/or environmental manipulations, on the morphology of the IMHV because they may mask other, more subtle, structural changes that occur as a result of behavioural experiences.

Aging↗

[3H]naloxone binding in the brain of the domestic chick (Gallus domesticus) determined by in vitro quantitative autoradiography.

The distribution of [3H]naloxone binding sites was investigated in the brains of one-day-old domestic chicks using quantitative receptor autoradiography. Among the forebrain regions, the greatest levels of opiate binding were found in hyperstriatum dorsale, hyperstriatum ventrale, hyperstriatum intercalatum supremum and neostriatum. Intermediate levels were found throughout the paleostriatal regions, septum, thalamus, archistriatum, hyperstriatum accessorium and area parahippocampalis whilst in hippocampus and ectostriatum the density of [3H]naloxone binding sites was low. In the hindbrain, high levels of opiate binding were found in optic tectum whereas in cerebellum the density of binding sites was barely above background. The greatest densities of opiate binding appear to coincide with regions found to be involved in sensory processing and memory storage.

Animals↗

A quantitative analysis of the synaptic development of the lobus parolfactorius of the chick (Gallus domesticus).

The lobus parolfactorius (LPO) of the chick has been shown to undergo an increase in the mean synaptic numerical density (Nvsyn) in response to one-trial passive avoidance learning (Stewart et al. 1987). The present study was undertaken in order to describe the pattern of normal development of synapses in the LPO, to further investigate the significance of this plastic response. The LPO's from each hemisphere of pre-hatch (16 days), and post-hatch (1 day, 9 day and 22 day old) chicks were processed for electron microscopy. Synapses were classified into asymmetric spine, asymmetric shaft, symmetric spine, and symmetric shaft synapses, on the basis of the density of the post-synaptic thickening and the nature of the post-synaptic target. A 3-dimensional stereological probe was used (the 'disector') to calculate Nvsyn, and mean projected height (Hsyn) of the post-synaptic density (PSD). Mean values for each age and hemisphere were compared with a 2-way analysis of variance test using paired samples. A six-fold increase in Nvsyn was seen between 16 days in ovo, and 9 days post-hatch. There was a hemispheric asymmetry at 9 days post-hatch, with the left hemisphere LPO containing 1.6 times as many synapses per micron-3. There was a subsequent period of reduction in synaptic density in the left hemisphere LPO between 9 and 22 days post-hatch. The Nv of all classes of synapse increased with age, but the proportions of the symmetrical synapses with respect to the total number of synapses, decreased with age. This decrease was of a similar magnitude for each hemisphere. A hemispheric difference was seen in post-hatch asymmetric synapses, with a greater proportion of asymmetric spine synapses in the left hemisphere. The magnitude of the hemispheric asymmetry was constant throughout the 3 week period of post-hatch development, but was not present in pre-hatch chicks. The PSD increased in length in each hemisphere by approximately 40% between post-hatch day 1 and post-hatch day 9. These data show that the LPO contains a synaptic population which undergoes substantial modification during the first week post-hatch. An asymmetry exists at post-hatch day 9 which is not present at the earlier ages investigated, nor indeed after 22 days post-hatch. This may have significance with regard to studies of passive avoidance learning in the one-day old chick, where an increase in both the size and number of synapses in the LPO has been demonstrated (Stewart et al. 1987).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Localization of GABA-like immunoreactivity in the ectostriatum of domestic chicks: GABA immunocytochemistry combined with Golgi impregnation.

The distribution of GABA-like immunoreactivity (GABA-LI) in the ectostriatal core (Ec) of domestic chicks (one to two days old) was investigated using (1) preembedding GABA immunocytochemistry and (2) Golgi impregnation and gold-toning combined with postembedding GABA immunocytochemistry. Two major classes of neurons which display GABA-LI were identified in chick Ec. Firstly, large GABA immunopositive cells which comprise at least two further subtypes: an ovoid or polygonal form of 14-18 microns diameter with no apparent polarity of dendrites and a smaller cell (10-14 microns) with ovoid or basket-shaped soma and often more polarized dendritic ramification. In both subtypes the dendritic surface is smooth or sparsely spiny. Secondly, a small GABA immunopositive cell which is characterized by a round cell body of 5-8 microns diameter and thin and sparsely ramifying dendrites of smooth surface or with irregular protrusions. Based upon comprehensive descriptions of ectostriatal cytoarchitectonics (Tömböl et al., 1988c), and synaptology (Watanabe et al., 1985), we argue that the GABA-immunopositive cell types of chick Ec are likely to represent inhibitory interneurons comparable with GABAergic inhibitory cell types described in mammalian visual cortex.

Animals↗

Adaptive changes in the central noradrenergic system in monocular deprived rats.

Noradrenaline and dopamine content, K+-stimulated release of 3H-noradrenaline, 3H-noradrenaline high-affinity uptake, 3H-prazosin and 3H-rauwolscine binding to alpha 1- and alpha 2-adrenoreceptors, respectively, were studied in isolated visual structures (visual cortex, superior colliculus, lateral geniculate nucleus) of three-month-old rats monocularly deprived by unilateral eyelid closure at postnatal day 11. Monocular deprivation resulted in a number of changes in comparison with normal littermates, mainly in the lateral geniculate nucleus, both ipsilateral and contralateral to the closed eye. K+-stimulated release of noradrenaline and high-affinity noradrenaline uptake were significantly increased, 3H-rauwolscine binding to alpha 2-adrenergic receptor sites was decreased in the contralateral and ipsilateral lateral geniculate nucleus. 3H-Prazosin binding to alpha 1-adrenergic receptors as well as the levels or noradrenaline and dopamine in the lateral geniculate nucleus were unaffected by monocular deprivation. The data support the suggestion that the lateral geniculate nucleus plays a key role in modulating visual information processing and that the lack of an adequate visual input within the first weeks of life interferes mainly with the development of the noradrenergic system at the level of the lateral geniculate nucleus.

Animals↗

Laminar pattern of cholinergic and adrenergic receptors in rat visual cortex using quantitative receptor autoradiography.

The laminar distribution of muscarinic acetylcholine receptors, including the M1-receptor subtype, of beta-adrenergic receptors, and noradrenaline uptake sites, was studied in the adult rat visual, frontal, somatosensory and motor cortex, using quantitative receptor autoradiography. In the visual cortex, the highest density of muscarinic acetylcholine receptors was found in layer I. From layer II/III to layer V binding decreases continuously reaching a constant binding level in layers V and VI. This laminar pattern of muscarinic receptor density differs somewhat from that observed in the non-visual cortical regions examined: layer II/III contained the highest receptor density followed by layer I and IV: lowest density was found in layer V and VI. The binding profile of the muscarinic cholinergic M1-subtype through the visual cortex shows a peak in cortical layer II and in the upper part of layer VI, whereas in the non-visual cortical regions cited the binding level was high in layer II/III, moderate in layer I and IV, and low in layer VI. Layers I to IV of the visual cortex contained the highest beta-adrenergic receptor densities, whereas only low binding levels were observed in the deeper layers. A similar laminar distribution was found also in the frontal, somatosensory and motor cortex. The density of noradrenaline uptake sites was high in all layers of the cortical regions studied, but with noradrenaline uptake sites somewhat more concentrated in the superficial layers than in deeper ones. The distinct laminar pattern of cholinergic and noradrenergic receptor sites indicates a different role for acetylcholine and noradrenaline in the functional anatomy of the cerebral cortex, and in particular, the visual cortex.

Animals↗

Training induced dendritic spine density changes are specifically related to memory formation processes in the chick, Gallus domesticus.

The density of dendritic spines on large, multipolar, projection neurons in the intermediate medial hyperstriatum ventrale (IMHV) of 1-day-old chicks was examined after training on a one-trial passive avoidance task. Chicks trained on the task were given a brief, subconvulsive transcranial electroshock 5 min after training, a procedure which rendered about half of them amnesic. The spine density was found to be significantly higher 24-26 h after training in the left IMHV in chicks that remembered the response compared to chicks rendered amnesic. These data, taken together with our earlier observations on spine density changes following training, argue strongly in favour of a specific role for dendritic spines in memory formation in the chick.

Animals↗

Quantitative autoradiographic analysis of the distribution of [3H]muscimol binding to GABA receptors in chick brain.

Quantitative receptor autoradiography was used to investigate the distribution of high-affinity GABA receptors (GABAA) in left and right hemispheres of the brains of 3-week-old chicks. The receptors were labelled with the potent GABA agonist [3H]muscimol. High levels of [3H]muscimol labelling were found throughout the fore-, mid-, and hindbrain, though considerable variation was found in different regions. In the telencephalon the highest concentration of specific binding was found in the hyperstriatum ventrale followed by the neostriatum, and then the lobus parolfactorius of the paleostriatal complex, whilst in the diencephalon highest levels of labelling were present in the infundibulum. In the midbrain distinct lamination was observed in the high levels of [3H]muscimol binding in the optic tectum and in the hind brain the highest density of labelling occurred in the granular layers of the cerebellum. Levels of labelling were generally low in the brainstem regions. The distribution of [3H]muscimol binding in the optic tectum and in the hind brain the highest density of labelling occurred in the granular layers of the cerebellum. Levels of labelling were generally low in the brainstem regions. The distribution of [3H]muscimol binding sites is in good agreement with our previous work on the distribution of GABA-immunoreactivity in the chick brain.

Animals↗

Changes in the number and structure of dendritic spines 25 hours after passive avoidance training in the domestic chick, Gallus domesticus.

One-day-old chicks spontaneously peck at a shiny chrome bead. If the bead is coated with methylanthranilate, a bitter tasting substance (M-chicks), they peck once and avoid a similar bead subsequently. Control chicks peck, and continue pecking at a bead dipped in water (W-chicks). Twenty-five hours after this one-trial passive avoidance training the brains were fixed and Golgi-impregnated. A class of large, multipolar, projection neurons from the intermediate medial hyperstriatum ventrale (IMHV) of both hemispheres from M- and W-chicks were examined for changes in their spine density and spine shape. An estimate of the true spine number was obtained using the correction formula of Feldman and Peters. M-chicks showed highly significant increases (P less than 0.0001) in spine density of between 89-113% in the left hemisphere, and 37-69% in the right, compared with W-chicks. There was a significant hemispheric asymmetry in W-chicks: the right hemisphere had approximately 47% more spines per micron than the left, and this difference was abolished after training. Following passive avoidance training, significant increases in spine head diameter (by approximately 9%) and decreases in spine stem length (by approximately 17%), with no significant alterations in overall spine length, were observed in the left hemisphere. The mean dendrite lengths were not significantly changed after training, but an asymmetry of this measure in W-chicks (left greater than right) was present in dendrite orders 2 (P less than 0.01) and 3 (P less than 0.02). These results show that spine densities can increase rapidly (within 25 h) following a one-trial passive avoidance training task and that spine shape changes can be found on the same dendrites which also show changes in spine number. The data support the view that dendritic spines are involved in memory formation processes.

Animals↗

gamma-Aminobutyric acid (GABA) immunoreactivity in mouse and rat first somatosensory (SI) cortex: description and comparison.

The location and morphological characteristics of gamma-aminobutyric acid (GABA)-immunopositive cells and their processes were studied in rat and mouse first somatosensory (SI) cortex (including 'barrels') in layer IV, and layers above (I-III), and below (V and VI). In coronal sections of SI cortex of both species GABA-immunopositive cells and punctate profiles were found in each of layers I-VI. The cells were of various sizes; the largest, located in layers III and V of each species, resemble the large basket cells seen in Golgi-impregnated material. Most of the immunopositive cells were multipolar and circular or ellipsoidal in shape, but occasionally bipolar cells with fusiform perikarya were also seen. In coronal sections, immunopositive cells did not form a characteristic pattern. GABA-immunopositive cells were observed to be most numerous in the supragranular layers whereas GABA-positive punctate profiles were more numerous in layer IV. In tangential sections from layer IV of SI cortex of both species, GABA-immunopositive cells, processes and punctate profiles were visible throughout the entire barrel field. The pattern of distribution of immunopositive cells was similar (a) in two different morphological groups--i.e. the posteromedial barrel subfield (PMBSF) and the anterolateral barrel subfield (ALBSF) in rat barrel field, and (b) in PMBSF barrels of both rat and mouse (excluding differences due to structural dissimilarities between rat and mouse barrels). GABA-immunopositive neurons were grouped mainly in the barrel side and septum and were visible frequently in small clusters. In barrels of both species GABA-immunopositive cells were of a variety of sizes and ranged in shape from ellipsoidal to circular.

Animals↗

The orientation of interneurones in the dorsal lateral geniculate nucleus of the rat: a quantitative study.

The orientation of the processes of 60 Golgi-impregnated interneurones in the rat dorsal lateral geniculate nucleus (dLGN) was studied quantitatively. A statistical analysis of the orientation of 238 interneuronal processes (dendrites and axon-like processes) showed that they were aligned preferentially along a dorsoventral axis through the dLGN and were predominantly oriented parallel with the outlying optic tract. Computer reconstructions of two of the Golgi-impregnated dLGN interneurones and their subsequent 3-dimensional computer rotations showed that their processes ramified in long columnar-shaped territories aligned dorsoventrally. There was little extension of these processes along the rostrocaudal axis of the nucleus. The data of this investigation provide evidence that the processes of 'inhibitory' interneurones in the rat dLGN are predominantly aligned along the dorsoventral axis, parallel with both the optic tract and with the afferent and efferent fibre tracts coursing through the nucleus.

Animals↗

Cell types of the hyperstriatum ventrale of the domestic chicken (Gallus domesticus): a Golgi study.

Two major classes of neurons were observed in Golgi impregnated material of two regions of the hyperstriatum ventrale (HV) (medial and rostral HV) of the chick telencephalon: a Golgi I type neuron with long axon (putative projection neuron) and a Golgi II type neuron with short axon (putative local circuit neuron). On the basis of the nature of the axon (bifurcating or non-bifurcating, ascending or descending) and of the dendrites (spiny or aspinous, and thickness), the following subclasses (types) were identified: two long axon (projection) and three short axon (local circuit) neurons in the medial (mid-telencephalic) hyperstriatum ventrale, and one major long axon (projection) and two short axon (local circuit) neurons in the rostral hyperstriatum ventrale. Possible patterns of intraregional connections between these neurons are suggested on the basis of cytoarchitectonics. The differences in cellular types and in internal connectivity patterns strongly suggest, in agreement with physiological and behavioural observations, that the medial (mid-telencephalic) hyperstriatum ventrale distinctly differs in cellular morphology from other parts of hyperstriatum ventrale in so far as it displays a greater diversity of neuronal types with an apparent higher degree of complexity.

Animals↗

Cell types of the paleostriatal complex of the domestic chicken (Gallus domesticus): a Golgi study.

A study of was carried out of Golgi impregnated material in two regions of the paleostriatal complex of the chick telencephalon, the lobus parolfactorius and paleostriatum augmentatum. As in our previous investigation of the chick medial hyperstriatum ventrale, two major classes of neurons were observed in the paleostriatum: a Golgi I type neuron (type 1) with long axon (putative projection neuron) and a Golgi II type neuron (type 2) with short axon (putative local circuit neuron). Based upon the nature of the axon (bifurcating or nonbifurcating, ascending or descending) and of the dendrites (spiny or aspinous, and thickness), the following subclasses (types) were identified: two projection and three local circuit neurons in the lobus parolfactorius and two projection and two local circuit neurons in the paleostriatum augmentatum. In addition to known differences in developmental chronology and extrinsic connectivity, the lobus parolfactorius clearly differs from palaeostriatum augmentatum in cellular composition and organization. From its cytoarchitectonic appearance we would suggest that the lobus parolfactorius is the paleostriatal region of greater complexity.

Animals↗

The structure of chicken ectostriatum. I. Golgi study.

Chicken ectostriatum was studied in Golgi preparations. In the two parts of the area (ectostriatum centrale and ectostriatum periphericum) the cell types were analysed. Projection neurons with long axon, and interneurons with short axon, could be distinguished in both parts of the region. The projection neurons are different in the two parts of the area, whilst the interneurons, with regard to their morphological features, are completely similar. The stellate-like projection neurons in ectostriatum centrale have 4-6 main dendrites, which ramify only slightly, and are densely covered by characteristic spines. The projection neurons in ectostriatum periphericum emit several (7-10) main dendrites, which have numerous bifurcations establishing a dense dendritic tree. The dendrites are densely covered by regular spines. The interneurons--in both parts of ectostriatum--have three subtypes. All have characteristic local axonal arborizations with varicose terminal sections. Some quantitative parameters of Golgi impregnated neurons were also measured and analysed by computer. Preliminary results of this study are included in the present publication.

Animals↗

GABAergic structures in the chick telencephalon: GABA immunocytochemistry combined with light and electron microscope autoradiography, and Golgi impregnation.

The distribution of gamma-aminobutyric acid (GABA)-ergic elements in 3 forebrain regions (medial mid-telencephalic hyperstriatum ventrale; paleostriatum augmentatum; lobus parolfactorius) of two-day-old domestic chicks was investigated using (1) light and electron microscope autoradiography following [3H]GABA uptake in vitro in combination with pre-embedding GABA immunocytochemistry and (2) Golgi impregnation and 'gold-toning' combined with postembedding GABA immunocytochemistry. In both the paleostriatal regions and the medial (mid-telencephalic) hyperstriatum ventrale, GABA immunolabelling was demonstrated with the pre-embedding technique. Radiolabelling with [3H]GABA was also shown in these regions, co-localised in many cases with the immunolabelling. In the paleostriatal regions, the majority of perikaryal labelling was found in ovoid, elongated or fusiform cell bodies of 6-7 micron diameter whereas in the medial (mid-telencephalic) hyperstriatum ventrale, larger (10-15 micron) multipolar and smaller (5-6 micron) bipolar neurons were found labelled. In the latter region, Golgi impregnated neurons of similar morphology were found to be immunopositive to GABA using the postembedding technique. The ultrastructure of [3H]GABA accumulating cells is characterised by pale or moderately granular nuclei with small invaginations, few mitochondria and a prominent Golgi apparatus. Astrocytes and ependymal cells are also labelled with [3H]GABA. GABA-labelled axon terminals represent 29-36% of the total in the 3 brain regions studied. They appear as electron-lucent boutons with few and often scattered synaptic vesicles and in most cases they form symmetrical axo-dendritic junctions.

Animals↗

Alterations in synaptic structure in the paleostriatal complex of the domestic chick, Gallus domesticus, following passive avoidance training.

A morphometric study was made of synapses in both left and right hemispheres of two regions of the chick paleostriatal complex, the paleostriatum augmentatum (PA) and the lobus parolfactorius (LPO), 24 h after passive avoidance training (methyl anthranilate, M-chicks), and in water-trained controls (W-chicks). The synaptic features examined were D, the mean length of the postsynaptic thickening; Nv.syn, the numerical density of synapses; Vv.syn, the volume density of the presynaptic bouton; V, the mean volume of the presynaptic bouton; Nv.ves, the numerical density of synaptic vesicles per bouton volume; ves.syn, the number of synaptic vesicles per presynaptic bouton; and K, the curvature of the synaptic contact zone. In the LPO there is a significant increase in the numerical density of synapses (Nv.syn) in both left and right hemispheres of M-compared with W-chicks (up to 59%, depending on the method of calculation used). A hemispheric asymmetry of postsynaptic thickening length (D) which is present in W-chicks (R greater than L by 10%) is reversed in M-chicks. There is no difference in the volume density of the presynaptic bouton (Vv.syn) or the mean bouton volume (V) either between W- and M-chicks, or between left and right hemispheres. Significant changes are found after avoidance training in both of the synaptic vesicle parameters measured. There is an increase of approximately 50%, both in the numerical density of synaptic vesicles (Nv.ves) and the number of vesicles per synaptic bouton (ves.syn), in the left hemisphere of M-chicks. No changes in the mean synaptic contact curvature (K) were observed after training, either of presynaptically concave, or presynaptically convex synapses, in either left or right hemispheres, nor did the percentage distribution of these different curvature classes vary greatly. In the PA there were no significant changes in D, Nv.syn or Vv.syn, either between M- and W-chicks, or between left and right hemispheres, 24 h after passive avoidance training. However, when the differences between the mean bouton volume (V) are examined, there is a significant increase in size of the boutons in the left hemisphere of M- compared to W-chicks. There is also a hemispheric asymmetry in both Nv.ves and ves.syn in W-chicks (R greater than L by approximately 15%) and this disappears on M-training. No changes in the mean synaptic contact curvature were observed after training, either of presynaptically concave, or presynaptically convex synapses, in either left or right hemispheres. However, the percentage distribution of these different curvature classes showed some variation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Autoradiographic localization of gamma-aminobutyric acid receptors in mouse barrel field.

The distribution of high-affinity gamma-aminobutyric acid (GABA) receptors in mouse posteromedial barrel subfield (PMBSF) in layer IV of the somatosensory (SI) cortex was studied using [3H]muscimol autoradiography in vitro. A qualitative study revealed a heterogeneous distribution in the density of [3H]muscimol binding in the barrel field. In the barrel sides and septum between the barrels [3H]muscimol binding exhibits the lowest level of labelling in the entire tissue. In comparison, [3H]muscimol binding in the hollows is considerably higher than in the barrel sides although the level is not homogeneous. These findings suggest that the barrel hollows are much richer in GABA receptors than the surrounding barrel sides and septa.

Animals↗

Ontogeny of 3H-muscimol binding to membranes of chick forebrain.

A potent GABA agonist, 3H-muscimol, was used to investigate the development of GABA receptors in left and right hemispheres of chick forebrain from day 12 in ovo to day 21 post-hatch. Total specific 3H-muscimol binding (p mol mg-1 protein) increases rapidly in ovo, reaching a peak at around day one post-hatch and then showing a slow decline to approximately 50% of the maximal level at day 21 post-hatch. Despite the considerable evidence from previous studies of lateralization of avian brain function, no significant hemispheric differences were found in 3H-muscimol binding (either of p mol hemisphere-1 or p mol mg protein-1) at any of the developmental ages examined.

Animals↗