Biomedical subjects

Douglas L Rosene

Publications and source records attributed to Douglas L Rosene.

At least 19 recordsLinked to original sources

PubMed2006

Maturational sequence of newly generated neurons in the dentate gyrus of the young adult rhesus monkey.

The generation of new neurons in the hippocampal dentate gyrus of adult mammals has been characterized in rodents, but the details of this process have not been described in the primate. Eleven young adult rhesus monkeys were given an injection of the DNA synthesis phase marker bromodeoxyuridine (BrdU) and killed at varying survival intervals (2 hours to 98 days). The immature neuronal marker TUC-4 (TOAD/Ulip/CRMP-4) was used to define three stages of morphological maturation. Stage I neurons had small somata and lacked dendrites. Stage II neurons had larger somata and short dendrites. Stage III neurons were similar in size to mature granule cells and had branching dendrites that extended into the molecular layer. Examination of TUC-4-positive immature neurons colabeled with BrdU indicated that stage I neurons first appeared 2 days after BrdU injection, stage II neurons at 14 days, and stage III neurons at 35 days. Electron microscopy of TUC-4-labeled cells showed that stage I cells had ultrastructural features of immature neurons, whereas stage III neurons were similar to mature granule cells and formed synapses in the molecular layer. This suggests that stage III neurons could potentially integrate into the circuitry of the dentate gyrus. This study shows that the maturational sequence for new neurons in the adult monkey is similar to that of the adult rodent; however, maturation takes a minimum of 5 weeks in the monkey, which is substantially longer than what has been reported in rodents.

Animals↗

PubMed2006

Frontal connections and cognitive changes in normal aging rhesus monkeys: a DTI study.

Recent anatomical studies have found that cortical neurons are mainly preserved during the aging process while myelin damage and even axonal loss is prominent throughout the forebrain. We used diffusion tensor imaging (DT-MRI) to evaluate the hypothesis that during the process of normal aging, white matter changes preferentially affect the integrity of long corticocortical association fiber tracts, specifically the superior longitudinal fasciculus II and the cingulum bundle. This would disrupt communication between the frontal lobes and other forebrain regions leading to cognitive impairments. We analyzed DT-MRI datasets from seven young and seven elderly behaviorally characterized rhesus monkeys, creating fractional anisotropy (FA) maps of the brain. Significant age-related reductions in mean FA values were found for the superior longitudinal fasciculus II and the cingulum bundle, as well as the anterior corpus callosum. Comparison of these FA reductions with behavioral measures demonstrated a statistically significant linear relationship between regional FA and performance on a test of executive function. These findings support the hypothesis that alterations to the integrity of these long association pathways connecting the frontal lobe with other forebrain regions contribute to cognitive impairments in normal aging. To our knowledge this is the first investigation reporting such alterations in the aging monkey.

Aging↗

PubMed2006

Age-related molecular reorganization at the node of Ranvier.

In myelinated axons, action potential conduction is dependent on the discrete clustering of ion channels at specialized regions of the axon, termed nodes of Ranvier. This organization is controlled, at least in part, by the adherence of myelin sheaths to the axolemma in the adjacent region of the paranode. Age-related disruption in the integrity of internodal myelin sheaths is well described and includes splitting of myelin sheaths, redundant myelin, and fluctuations in biochemical constituents of myelin. These changes have been proposed to contribute to age-related cognitive decline; in previous studies of monkeys, myelin changes correlate with cognitive performance. In the present study, we hypothesize that age-dependent myelin breakdown results in concomitant disruption at sites of axoglial contact, in particular at the paranode, and that this disruption alters the molecular organization in this region. In aged monkey and rat optic nerves, immunolabeling for voltage-dependent potassium channels of the Shaker family (Kv1.2), normally localizing in the adjacent juxtaparanode, were mislocalized to the paranode. Similarly, immunolabeling for the paranodal marker caspr reveals irregular caspr-labeled paranodal profiles, suggesting that there may be age-related changes in paranodal structure. Ultrastructural analysis of paranodal segments from optic nerve of aged monkeys shows that, in a subset of myelinated axons with thick sheaths, some paranodal loops fail to contact the axolemma. Thus, age-dependent myelin alterations affect axonal protein localization and may be detrimental to maintenance of axonal conduction.

Aging↗

PubMed2006

Asymmetry of neuron numbers in the hippocampal formation of prenatally malnourished and normally nourished rats: a stereological investigation.

There is considerable evidence for lateralization of hippocampal function and hemispheric asymmetry in humans. In the rat, studies have reported asymmetries in the thicknesses of layers, the volumes of hippocampal subfields, and the density of cells at specific points along the septotemporal axis. To determine if there is an asymmetry of neuron numbers and whether prenatal malnutrition affects any asymmetries, 90-day old male Sprague-Dawley rats that were either normally nourished or malnourished prenatally were perfused with 4% paraformaldehyde and the brains cut into 30-micro m sections. One interrupted series of sections through the entire hippocampus was analyzed stereologically to estimate the total number of neurons in the hilus of the dentate gyrus, the CA3/CA2 stratum pyramidale (SP), the CA1 SP, and the SP of the prosubiculum/subiculum of both hemispheres. Significant asymmetries (P < 0.05) were found in the CA1 and CA3/CA2 subfields, with the right hemisphere containing 21 and 6% fewer neurons, respectively. Malnutrition reduced neuron numbers in the CA1 subfield by 12%, but did not alter the hemispheric asymmetry. Our findings agree with previous reports of left dominant asymmetries in the rat brain and suggest that this may result from differences in total numbers of neurons.

Analysis of Variance↗

PubMed2005

Executive system dysfunction occurs as early as middle-age in the rhesus monkey.

As our understanding of age-related cognitive decline advances, studies are now focusing on identification of those areas of cognitive function that undergo the first changes with age. In the present study, in order to determine whether executive function is sensitive to the aging process, we assessed the performance of 16 monkeys of middle-age (12-19 years of age) on the conceptual set-shifting task, an analogue of the Wisconsin Card Sorting Test (WCST). We compared their performance to that of seven young adult (5-9 years of age) and 18 aged monkeys (20-30). The findings showed that middle-aged monkeys, like those of advanced age, were significantly impaired on the conceptual set-shifting task (CSST). These findings parallel those of recent studies in humans demonstrating an increase in perseverative errors on the WCST by middle-aged as well as aged individuals and, in turn, support the notion that disruption of executive function is one of the earliest changes in cognition to occur in normal aging.

Age Factors↗

PubMed2005

A non-human primate test of abstraction and set shifting: an automated adaptation of the Wisconsin Card Sorting Test.

Functional assessment of the prefrontal cortices in the non-human primate began with the seminal work of Jacobsen in the 1930s. However, despite nearly 70 years of research, the precise nature of the cognitive function of this region remains unclear. One factor that has limited progress in this endeavor has been the lack of behavioral tasks that parallel most closely those used with humans. In the present study, we describe a test for the non-human primate that was adapted from the Wisconsin Card Sorting Task (WCST), perhaps the most widely used test of prefrontal cognitive function in humans. Our adaptation of this task, the Conceptual Set-Shifting Task (CSST), uses learning criteria and stimuli nearly identical to those of the WCST. The CSST requires the animal to initially form a concept by establishing a pattern of responding to a given stimulus class, maintain responding to that stimulus class, and then shift to a different stimulus class when the reward contingency changes. The data presented here establishes baseline performance on the CSST for young adult rhesus monkeys and demonstrates that components of prefrontal cognitive function can be effectively assessed in the non-human primate in a manner that parallels the clinical assessment of humans.

Animals↗

PubMed2005

A statistically based density map method for identification and quantification of regional differences in microcolumnarity in the monkey brain.

We present a statistical density map method derived from condensed matter physics to quantify microcolumns, the fundamental computational unit of the cerebral cortex. This method provides measures for microcolumnar strength, width, spacing, length, and periodicity. We applied this method to Nissl-stained 30 microm thick frozen sections from areas 46, TE, and TL of rhesus monkey brains, areas that differ visually in microcolumnarity and are associated with different cognitive functions. Our results indicate that microcolumns in these areas are similar in width, spacing, and periodicity, but are stronger (possess a higher neuronal density) in area TE, as compared to areas TL and 46. We modeled the effect of section orientation on microcolumnar spacing and demonstrated that this method provides an adequate estimate of spacing. We also modeled disruption of microcolumnarity by performing simulations that randomly displace neurons and demonstrated that displacements of only one neuronal diameter effectively eliminate microcolumnar organization. These results indicate that our density map method is sensitive enough to detect and quantify subtle differences in microcolumnar organization that may occur in the context of development, aging, and neuropathology, as well as between areas and species.

Animals↗

PubMed2005

Cognitive impairment in aged rhesus monkeys associated with monoamine receptors in the prefrontal cortex.

The "frontal aging hypothesis" has been proposed by many researchers suggesting that the earliest and most severe age-related changes in the cortex occur in the frontal lobes. Two of these changes include decreases in cognitive functions mediated by the prefrontal cortex (PFC) and significant decreases in norepinephrine (NE) and dopamine (DA). To investigate whether the changes in these neurotransmitter systems are directly related to the cognitive decline seen in aging we utilized the rhesus monkey as a model of normal human aging. Our goal was to determine if age-related changes in cognition is associated with changes in norepinephrine and dopamine receptor binding density in the PFC. Eight young monkeys between five and ten years of age (six males and two female) and eight aged monkeys between 25 and 32 years of age (five males and three females) were behaviorally characterized. Subsequently on-the-slide in vitro binding assays were used to quantify the alpha-1 adrenergic, alpha-2 adrenergic and DA1 receptors as well as the NE and DA uptake receptors. Aged animals as a group demonstrated significant cognitive impairments and aging produced a significant decrease in alpha-1 adrenergic and alpha-2 adrenergic receptor binding in the PFC but no significant change in binding for the DA1 receptor or the NE or DA uptake receptors. Further analysis revealed a significant relationship between monoamine receptor binding and cognitive performance on three tasks: delayed non-matching to sample, delayed recognition span test and the conceptual set-shifting task.

Adrenergic alpha-Agonists↗

PubMed2005

Effect of prenatal protein malnutrition on numbers of neurons in the principal cell layers of the adult rat hippocampal formation.

Malnutrition has been associated with a variety of functional and anatomical impairments of the hippocampal formation. One of the more striking of these is widespread loss of hippocampal neurons in postnatally malnourished rats. In the present study we have investigated the effect of prenatal malnutrition on these same neuronal populations, neurons that are all generated during the period of the dietary restriction. In prenatally protein deprived rats, using design-based stereology, we have measured the regional volume and number of neurons in the hilus of the dentate gyrus and the pyramidal cell layers of CA3, CA2, CA1, and the subiculum of 90-day-old animals. These results demonstrated a statistically significant reduction of 20% in neuron numbers in the CA1 subfield, while numbers in the other subfields were unchanged. There was a corresponding significant reduction of 22% in the volume of the CA1 subfield and a significant 14% decrease in the volume of the pyramidal layer of the subiculum. The change in volume of the pyramidal layer of the subiculum without neuron loss may reflect loss of CA1 afferent input to the pyramidal layer. Although the effect of nutritional deprivation on the neuronal population appears to be different in pre- and postnatal malnutrition, both dietary paradigms highlight the vulnerability of key components of the hippocampal trisynaptic circuit (consisting of the dentate granule cell mossy fibers projection to CA3 pyramids and the CA3 projection to the CA1 pyramids), which is an essential circuit for memory and learning.

Animals↗

PubMed2005

Sex, age, and training modulate spatial memory in the rhesus monkey (Macaca mulatta).

The authors tested 90 rhesus monkeys (Macaca mulatta) on a task of spatial memory, the spatial Delayed Recognition Span Test. The results showed that performance declined significantly with age, males had greater scores than females, and the rate of apparent decline with age was greater in males than in females. Both working and reference memory declined with age, but only working memory showed sex differences. The authors compared these data with that of 22 monkeys who were trained on a simpler version of the task before formal testing. Training had no effect on males but dramatically improved working memory in young females. The results confirm a male advantage in spatial working memory at a young age and confirm a greater decline with age in males than in females. It is important to note that prior training completely reverses the deficits of young females.

Age Factors↗

PubMed2005

Increased action potential firing rates of layer 2/3 pyramidal cells in the prefrontal cortex are significantly related to cognitive performance in aged monkeys.

The neurobiological substrates of significant age-related deficits in higher cognitive abilities mediated by the prefrontal cortex (PFC) are unknown. To address this issue, whole-cell current-clamp recordings were used to compare the intrinsic membrane and action potential (AP) firing properties of layer 2/3 pyramidal cells in PFC slices from young and aged behaviorally characterized rhesus monkeys. Most aged subjects demonstrated impaired performance in Delayed Non-Match to Sample (DNMS) task acquisition, DNMS 2 min delay and the Delayed Recognition Span task. Resting membrane potential and membrane time constant did not differ in aged relative to young cells, but input resistance was significantly greater in aged cells. Single APs did not differ in terms of threshold, duration or rise time, but their amplitude and fall time were significantly decreased in aged cells. Repetitive AP firing rates were significantly increased in aged cells. Within the aged group, there was a U-shaped quadratic relationship between firing rate and performance on each behavioral task. Subjects who displayed either low or very high firing rates exhibited poor performance, while those who displayed intermediate firing rates exhibited relatively good performance. These data indicate that an increase in AP firing rate may be responsible, in part, for age-related PFC dysfunction.

Action Potentials↗

PubMed2005

Light and electron microscopic immunohistochemical detection of bromodeoxyuridine-labeled cells in the brain: different fixation and processing protocols.

Bromodeoxyuridine (BrdU) immunohistochemistry is the method of choice for labeling newly generated cells in the brain. Most BrdU studies utilize paraformaldehyde-fixed brain tissue because of its compatibility with both BrdU and other immunohistochemical methods. However, stronger fixation is required for electron microscopic studies, and unfixed tissue is needed for biochemical and molecular studies. Because there are no systematic studies comparing the effects of different fixatives on BrdU immunohistochemistry in brain tissue, we compared BrdU immunohistochemical methods in brain tissue fixed with 4% paraformaldehyde, a mixed glutaraldehyde-paraformaldehyde fixative for electron microscopy, and unfixed tissue from brains perfused only with buffer and flash frozen. After optimizing immunostaining protocols, qualitative assessments of light microscopic diaminobenzidine labeling and of double-label immunofluorescence with confocal microscopy demonstrated excellent BrdU labeling in each of the three groups. Quantitative stereological assessment of the number of BrdU-labeled cells in rat dentate gyrus showed no significant difference in the number of labeled cells detected with each perfusion protocol. Additionally, we developed a protocol to visualize BrdU-labeled cells in the electron microscope with adequate preservation of fine structure in both rat and monkey brain.

Animals↗

PubMed2004

Age-related reduction in microcolumnar structure in area 46 of the rhesus monkey correlates with behavioral decline.

Many age-related declines in cognitive function are attributed to the prefrontal cortex, area 46 being especially critical. Yet in normal aging, studies indicate that neurons are not lost in area 46, suggesting that impairments result from more subtle processes. One cortical feature that is functionally important, but that has not been examined in normal aging because of a lack of efficient quantitative methods, is the vertical arrangement of neurons into microcolumns, a fundamental computational unit of the cortex. By using a density-map method derived from condensed-matter physics, we quantified microcolumns in area 46 of seven young and seven aged rhesus monkeys that had been cognitively tested. This analysis demonstrated that there is no age-related reduction in total neuronal density or in microcolumn width, length, or periodicity. There was, however, a statistically significant decrease in the strength of microcolumns, indicating microcolumnar disorganization. This reduction in strength was significantly correlated with age-related cognitive decline on tests of spatial working memory and recognition memory independent of the effect of age. Modeling demonstrated that random neuron displacements of approximately 30% of a neuronal diameter (<3 mum) produced the observed reduction in strength. Hence, it is possible that, with changes in dendrites and myelinated axons, subtle displacements of neurons occur that alter microcolumnar structure and correlate with age-induced dysfunction. Therefore, quantitative measurement of microcolumnar structure may provide a sensitive morphological method to assay microcolumnar function in aging and other conditions.

Aging↗

PubMed2004

Motor projections to the basis pontis in rhesus monkey.

Motor corticopontine studies suggest that the pons is topographically organized, but details remain unresolved. We used physiological mapping in rhesus monkey to define subregions in precentral motor cortex (M1), injected isotope tracers into M1 and the supplementary motor area (SMA), and studied projections to the basis pontis. Labeled fibers descend in the internal capsule (SMA in anterior limb and genu; M1 in posterior limb) and traverse the midsection of the cerebral peduncle, where SMA fibers are medial, and face, arm, and leg fibers are progressively lateral. Each motor region has unique terminations in the ipsilateral basis pontis and nucleus reticularis tegmenti pontis. Projections are topographically organized, preferentially in the caudal half of the pons, situated in close proximity to traversing corticofugal fibers. In nuclei that receive multiple inputs, terminations appear to interdigitate. Projections from the SMA-face region are most medial and include the median pontine nucleus. M1-face projections are also medial but are lateral to those from SMA-face. Hand projections are in medially placed curved lamellae in mid- and caudal pons. Dorsal trunk projections are in medial and ventral locations. Ventral trunk/hip projections encircle the peduncle in the caudal pons. Foot projections are heaviest caudally in laterally placed, curved lamellae. These results have relevance for anatomical clinical correlations in the human basis pontis. Furthermore, the dichotomy of motor-predominant caudal pons projections to cerebellar anterior lobe, contrasted with associative-predominant rostral pons projections to cerebellar posterior lobe, is consistent with new hypotheses regarding the cerebellar contribution to motor activity and cognitive processing.

Animals↗

PubMed2004

Effects of prenatal protein malnutrition and acute postnatal stress on granule cell genesis in the fascia dentata of neonatal and juvenile rats.

Although postnatal genesis of granule cells in the hippocampal fascia dentata is known to be influenced by prenatal protein deprivation or by stress, the combined effects of prenatal protein malnutrition and stress on these cells are unknown. This study was designed to examine this combined effect. Well-nourished and prenatally malnourished pups on postnatal day 7 (P7) were stressed by maternal separation and reduction of body temperature and on postnatal day 30 (P30) by immobilization with restraint. Bromodeoxyuridine (BrDU) was injected at the time of stress, and 2 h later, the numbers of immunolabeled cells were quantified by standard stereological techniques. In comparison to controls, prenatally malnourished rats showed a significantly lower number of cells tagged in the fascia dentata on P7 (p < or =0.05), and a significantly higher number of cells (p < or =0.05) on P30. In both age groups, control rats exposed to acute stress showed a significantly decreased number of cells tagged in the fascia dentata (p < or =0.05). In contrast, neurogenesis in malnourished rats was not significantly affected by acute stress at either age. Thus, the pattern of neurogenesis in the fascia dentata and its response to stress has been fundamentally altered by prenatal protein deprivation.

Acute Disease↗

PubMed2004

Ataxia after pontine stroke: insights from pontocerebellar fibers in monkey.

Basis pontis lacunes cause contralateral but rarely ipsilateral ataxia. We explored this phenomenon with isotope tract tracing in the rhesus monkey. Labeled pontocerebellar fibers cross midline and disperse widely in the opposite hemipons before coalescing in the brachium pontis. This anatomical arrangement suggests that small pontine strokes spare sufficient decussating pontocerebellar fibers to prevent ipsilateral dysmetria, and that ipsilateral dysmetria after large pontine stroke represents a disconnection syndrome.

Animals↗

PubMed2004

Beta-secretase activity increases with aging in human, monkey, and mouse brain.

Amyloid beta protein (A beta) accumulates in the brains of aging humans, amyloid precursor protein (APP) transgenic mouse lines, and rhesus monkeys. We tested the hypothesis that aging was associated with increased activity of the beta-site amyloid precursor protein cleaving enzyme (beta-secretase, BACE) in brain. We evaluated BACE activity, BACE protein, and formic acid-extractable A beta levels in cohorts of young (4 months old) and old (14 to 18 months old) nontransgenic mice (n = 16) and Tg2576 APP transgenic mice (n = 17), young (4.4 to 12.7 years old) and old (20.9 to 30.4 years old) rhesus monkeys (n = 17), and a wide age range (18 to 92 years old) of nondemented human brains (n = 25). Aging was associated with increased brain A beta levels in each cohort. Furthermore BACE activity increased significantly with age in mouse, monkey, and human brains, independent of brain region. BACE protein levels, however, were unchanged with age. BACE activity correlated with formic acid-extractable A beta levels in transgenic mouse, nontransgenic mouse, and human cortex, but not in monkey brain. These data suggest that an age-related increase of BACE activity contributes to the increased production and accumulation of brain A beta, and potentially predisposes to Alzheimer's disease in humans.

Aging↗

PubMed2003

Parcellation of cortical afferents to three distinct sectors in the parahippocampal gyrus of the rhesus monkey: an anatomical and neurophysiological study.

The posterior parahippocampal gyrus (PHG) of the rhesus monkey (Macaca mulatta) is comprised of three distinct cortical areas based on cytoarchitecture, connectivity, and neurophysiological response properties. Fluorescent retrograde tracers placed in each PHG area demonstrated unique patterns of cortical afferent input to areas TH, TL, and TF. Area TF receives inputs from the multimodal cortices of the superior temporal sulcus including areas PGa, TPO, and MST, from the visuospatial parietal area PG-Opt, and from visual areas V3A and dorsal V4. Area TL receives afferents from the inferotemporal region including visual areas TE1 and TE2 as well as from areas TEa, IPa, and FST in the lower bank and depth of the superior temporal sulcus. In contrast, the input to area TH is from the rostral part of superior temporal gyrus, including the auditory association areas TS1-3, and from the middle sector of area TPO in the superior temporal sulcus. Frontal and cingulate areas also project to the PHG in largely differential patterns. To further investigate this a correlative electrophysiological study of the three PHG areas resulted in a confirmation of these differential cortical inputs such that visually responsive neurons were found in areas TF and TL, auditory responsive neurons or bimodal auditory/visual-responsive neurons in area TH, and somatosensory-responsive neurons at the TF/TL border. Since each PHG area also receives differential hippocampal input, these data suggest that the processing of unimodal or multimodal information may be related to memory processing functions that are largely segregated within areas TH, TL, and TF.

Afferent Pathways↗