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Biomedical subjects

W T Greenough

Publications and source records attributed to W T Greenough.

At least 19 recordsLinked to original sources

5B4-CAM expression parallels neurite outgrowth and synaptogenesis in the developing rat brain.

In order to study whether 5B4-CAM expression parallels neurite outgrowth and synaptogenesis, a monoclonal antibody, 5B4, was used, which recognizes both fetal (185-250 kD) and adult (140 kD, 180 kD) forms of the neural cell adhesion molecule (N-CAM), to identify and localize the antigen in rat tissue during developmental ages P1 through P31 and in adults between P60 and 2 years of age. A ubiquitous pattern of intense immunolabelling was detected during the earliest stages of development. 5B4-CAM expression paralleled process outgrowth and the early stages of synaptogenesis in the cerebral cortex, hippocampal formation, and cerebellum. In the adult, immunoreactivity was generally less intense, but the cerebral cortex and hippocampal and cerebellar molecular layers, all areas implicated in learning-associated plasticity, retained substantial immunoreactivity. The inner one-third of the dentate gyrus molecular layer, an area implicated in axonal sprouting and reactive synaptogenesis, was particularly intensely labelled. Evidence from this work suggests that 5B4-CAM expression may be useful in monitoring neurite outgrowth and the early stages of synapse formation during development and possibly axonal sprouting and reactive synaptogenesis in the adult.

Animals

Increases in dendritic length in occipital cortex after 4 days of differential housing in weanling rats.

To assess the capacity for experience to induce rapid alterations in the dendritic fields of cortical neurons, male Long-Evans hooded rats aged 30-31 days were housed in either a complex environment (EC) or an individual cage (IC) for 4 days. The basilar dendrites of layer III pyramidal cells in area 17 of visual cortex were measured in Golgi-stained sections. EC rats exhibited significant increases in total dendritic length and total number of branches. This finding demonstrates that the structural modifications previously reported after 30 days in the complex environment are well underway after only 4 days.

Animals

Exercise and the brain: angiogenesis in the adult rat cerebellum after vigorous physical activity and motor skill learning.

This study compared the morphology of cerebellar cortex in adult female rats exposed for 1 month to repetitive exercise, motor learning, or an inactive condition. In the exercise conditions, rats that were run on a treadmill or housed with access to a running wheel had a shorter diffusion distance from blood vessels in the molecular layer of the paramedian lobule when compared to rats housed individually or rats that participated in a motor skill learning task. Rats taught complex motor skills substantially increased the volume of the molecular layer per Purkinje neuron and increased blood vessel number sufficiently to maintain the diffusion distance. These results dissociate angiogenesis associated with increased neuropil volume (as seen in the motor learning group) from angiogenesis associated with increased metabolic demands (as seen in the exercise groups). While the volume fraction of mitochondria did not differ among groups, the mitochondrial volume fraction per Purkinje cell was significantly increased in the motor skill rats. This appears to parallel the previously reported increase in synapses and associated neuropil volume change.

Animals

Plasticity of GFAP-immunoreactive astrocyte size and number in visual cortex of rats reared in complex environments.

Previous studies have indicated effects of postweaning rearing environment complexity on astrocyte nuclei. This study examined the effects of rearing for 10, 30 or 67 days in a complex (EC), social (SC) or individual cage (IC) environment upon glial fibrillary acidic protein (GFAP) immunoreactive astrocytes of the occipital cortex. EC rats exposed to their environment for 30 days or longer had a greater surface density of astrocytic processes (Sv) than SC or IC rats. The greater surface density of EC cortical astrocytes appeared to be due to an increase in the mean size of astrocytes after 30 days of differential environmental exposure. After 67 days of environmental exposure, however, the greater Sv appeared to be due to an increase in the number of astrocytes. Astrocytic plasticity appears to develop rather slowly during exposure to a complex environment and appears to involve two stages. The first stage is a hypertrophy of existing astrocytes and the second stage involves proliferation or retarded death of astrocytes. These changes may be related to brain information processing since astrocytes are known modulators of synaptic activity and may possibly serve as regulators of synaptic density.

Analysis of Variance

Capillary and mitochondrial support of neural plasticity in adult rat visual cortex.

Young adult rats (60 days old) were placed in complex environments (EC) or kept in individual cages (IC) for 10, 30, or 60 days. Previously reported findings in these same animals of synaptogenesis, decreased neuronal density, and increased cortical thickness in the EC animals demonstrated that cortical volume substantially expanded after 30 days. Such expansion would have spread apart existing capillaries and mitochondria, thereby diluting metabolic support. However, capillary spacing and mitochondrial volume fraction were maintained in these EC animals after 30 days, suggesting that new capillaries and mitochondria had infiltrated the tissue. Furthermore, many small vessels appeared after 10 days of complex experience, followed by expansion in vessel size until vessels from rats in EC for 60 days were larger than those from rats in IC for 60 days. The findings of constant vessel spacing in the face of expanding tissue volume, along with a set of small vessels that subsequently increased in size, suggest that small-sized new vessels were introduced in EC cortex by 10 days but had not matured in size until after 30 days. The results indicate that young adult rats can generate new capillaries and mitochondria in response to increased metabolic demands, but in a less vigorous fashion than in previously described weanling animals.

Animals

Astrocyte hypertrophy in the dentate gyrus of young male rats reflects variation of individual stress rather than group environmental complexity manipulations.

Glial hypertrophy is associated with synaptogenesis in visual cortex and with stress-induced damage in the hippocampus. This study examined astrocytes in the dentate gyrus of male weanling rats exposed to complex or standard laboratory environments. No group differences in astrocytic surface density were observed, as expected in this brain region where group differences in synaptogenesis in male rats are reportedly minimal. Similarly, no group differences in adrenal weight were observed. Across all treatment groups, however, a significant positive correlation (r = 0.57) between adrenal weight and surface density of astrocytic processes was found. Considerable variation in responses of individual rats to their environments occurs in both the complex and the laboratory cage environments, and animals responding poorly may have had heavier adrenals and greater astrocyte reactivity in the dentate gyrus. Thus astrocyte hypertrophy in the dentate gyrus reflects the stress history of the individual rat and not any differential effects of rearing in a complex or a laboratory cage environment.

Adrenal Glands

Effect of environmental complexity on size of the superior colliculus.

The present study examined effects of environmental complexity on the size of the superior colliculus, a subcortical structure involved in visuomotor functions. Long-Evans hooded rats raised together in a complex environment for 48 weeks were compared with their littermates housed in individual cages. The depth and area of the superficial gray layer of the superior colliculus were about 5-6% greater in the group from the complex environment, while the deeper layers of the superior colliculus showed no significant differences. The magnitude of the differences approached those reported for the neocortex, which has been considered to be distinctive in its morphological responsiveness to differential environmental complexity. The findings also extend previous observations that visual deprivation leads to shrinkage of the superficial gray layer and indicate that the morphology of this subcortical visual area is responsive to varying degrees of environmental stimulation.

Animals

Learning causes synaptogenesis, whereas motor activity causes angiogenesis, in cerebellar cortex of adult rats.

The role of the cerebellar cortex in motor learning was investigated by comparing the paramedian lobule of adult rats given difficult acrobatic training to that of rats that had been given extensive physical exercise or had been inactive. The paramedian lobule is activated during limb movements used in both acrobatic training and physical exercise. Acrobatic animals had greater numbers of synapses per Purkinje cell than animals from the exercise or inactive groups. No significant difference in synapse number or size between the exercised and inactive groups was found. This indicates that motor learning required of the acrobatic animals, and not repetitive use of synapses during physical exercise, generates new synapses in cerebellar cortex. In contrast, exercise animals had a greater density of blood vessels in the molecular layer than did either the acrobatic or inactive animals, suggesting that increased synaptic activity elicited compensatory angiogenesis.

Animals

Perfusion method for preparing pig brain cortex for Golgi-Cox impregnation.

The perfusion procedure described in this paper produces high quality impregnation of pig visual and somatosensory cortical neurons with a Golgi-Cox solution. Starting within 30 min after death, pig heads were perfused with a fixative solution composed of a mixture (v/v) of liquid phenol, 5%; formalin, 14%; ethylene glycol, 25%; methanol, 28%; and water, 28% for two periods of 4 hr each. After perfusion, the heads were chilled for at least 18 hr. The entire brain was removed from the skull and then placed in 10% buffered formalin, where it remained for at least 10 days before taking the blocks that were to be immersed in the Golgi-Cox solution. Three weeks spent in the Golgi-Cox solution typically produced uniform neuron impregnation. The tissue blocks were then embedded in celloidin and sectioned at 120 micron. This procedure avoids the following difficulties: Golgi-Cox methods that produced excellent results with rodent or primate tissue were unsuccessful with pig tissue, placing fresh tissue in Golgi-Cox solution resulted in incomplete neuron impregnation, and immersion fixation in 10% buffered formalin without perfusion resulted in excessive staining of glia.

Animals

Complex experience promotes capillary formation in young rat visual cortex.

The metabolic support of neural plasticity was examined by comparing cerebral vasculature of weanling rats reared in complex environments (EC) to littermates reared individually (IC) or socially in pairs (SC). EC rats have a thicker occipital cortex, more synaptic contacts per neuron and larger dendritic arbors compared to SC or IC rats, potentially increasing local metabolic demands on microvasculature. Capillaries of EC rats were closer together than those of SC or IC rats and potentially filled a greater fraction of cortex with blood. The closer capillary spacing in young EC rats suggests compensatory angiogenesis in response to increased metabolic demand.

Animals

Effects of unilateral and bilateral training in a reaching task on dendritic branching of neurons in the rat motor-sensory forelimb cortex.

Effects of motor training on a neocortical nerve cell population involved in performance of the motor task were assessed by measuring Layer V pyramidal neuron apical dendritic branching in motor-sensory forelimb cortex of rats trained to reach into a tube for food. Rats were trained to reach with the forepaw they preferred to use (group PRAC), the nonpreferred forepaw (REV), both forepaws (ALT), or neither forepaw (CONT). Across groups, hemispheres opposite trained forepaws had larger apical dendritic fields, in terms of total dendritic length, number of oblique branches from the apical shaft, and length of terminal branches. Similar, although somewhat less consistent, effects were seen when results were analyzed for between- (CONT vs ALT) and within-subject comparisons (trained vs nontrained hemispheres of REV and PRAC). This finding is compatible with the hypothesis that altered dendritic patterns, with associated synapses, are involved in storage of information from the training experience. The within-subject effects mitigate suggestions that these differences arise from generally acting hormonal or metabolic consequences of the training experience, although the possibility that these effects result from neural activity per se and are unrelated to information storage cannot be excluded.

Animals

Cerebellar plasticity: modification of Purkinje cell structure by differential rearing in monkeys.

Dendritic branching in Purkinje and granule cells and the diameters of Purkinje cell somas were compared in several cerebellar areas of monkeys reared in isolation, with social experience, or in a large colony. In the colony-reared monkeys, spiny branchlets of Purkinje cells were more extensive in the paraflocculus and the nodulus than they were in the other two groups. Granule cell dendritic branching in the paraflocculus and nodulus did not differ across groups. In addition, Purkinje cell somas were larger in the uvula and the nodulus of the colony animals than in the other groups. These data indicate that the social and physical environment during development influences the morphology of cerebellar Purkinje cells.

Animals

A quantitative investigation of spine and dendrite development of neurons in visual cortex (area 17) of Macaca nemestrina monkeys.

In a previous Golgi study (Lund et al., '77) which examined the development of the macaque monkey striate cortex (area 17) it was observed that the dendrites of neurons within the visual cortex show a marked increase in the number of spines on their surface during the first eight weeks of postnatal life. The qualitative observation was also made that all neurons then showed a marked decrease in spine numbers by the time the animal was adult. Since these spines are known to be sites of synaptic contact, changes in their numbers may reflect changes in synapse populations on these neurons. This study examines quantitatively spine frequency and total dendritic development of Golgi impregnated neurons in monkeys ranging in age from 145 days gestation to adult. Four cell types were studied: spiny stellate neurons from laminae IVCalpha and IVCbeta and pyramidal neurons with soma in either lamina IIIB or upper lamina VI. After consideration of possible sources of variation in spine numbers several conclusions are made: (1) Dendritic spine development appears to be a tightly controlled process both in terms of actual numbers of spines on a neuron at any one age and in the rate of change of spine frequency. (2) The neurons populations examined all show a gradual increase in spine numbers up to eight weeks of age. (3) At least two different trends are found in spine population maturation after the eight week point: (A)-the spine population may remain constant at the eight week level for same period of time or (B)-there may be a rapid decline in spine numbers following the eight week peak. (4) There is a suggestion that those neurons associated with direct input, or early stages in the relays, from the parvocellular geniculate laminae show trend B, while those associated with magnocellular input, or later order combined relays within the cortex, show trend A. (5) Different parts of a single pyramidal neuron dendrite may show either trend A or trend B, depending on the lamina location of the dendritic segment considered. (6) All neurons show spine population decreases between nine months of age and adult (5-7 years) suggesting continuing long term maturational changes.

Animals

Subsynaptic plate perforations: changes with age and experience in the rat.

The relative frequency of appearance of discontinuities in the postsynaptic thickening, or perforations in the subsynaptic plate, increased with age and experience. Rats reared from weaning in complex or social environments had a significantly higher proportion of occipital cortical synapses with perforations than did rats reared in isolation. In addition, the relative frequency of these perforations more than tripled between 10 and 60 days of age. Shifts in the frequency of perforations can occur independently of changes in the size of synpases. This result suggests a new potential mechanism of synaptic plasticity.

Aging

Sex differences in dentritic patterns in hamster preoptic area.

Sexual dimorphism is described in the dentritic field pattern of Golgi-stained neurons from the dorsomedial preoptic area of adult golden hamsters (Mesocricetus auratus). Data were obtained through a mathematical reconstruction of dentritic densities of neurons sampled from this area in males and females. Males tended to have a central concentration, while females showed an irregular dendritic density distribution with concentrations dorsolateral, ventral and medial to the area of highest dentriic density in the males. These results suggest sex differences in the afferent inputs to neurons in the dorsomedial preoptic area which may be related to functional sexual dimorphism in physiology and behavior.

Animals