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In rhesus macaques, previous studies have shown that episodic exposure to allergen alone or combined with ozone inhalation during the first 6 months of life results in a condition with many of the hallmarks of asthma. This exposure regimen results in altered development of the distal airways and parenchyma (Avdalovic et al., 2012). We hypothesized that the observed alterations in the lung parenchyma would be permanent following a long-term recovery in filtered air (FA) housing. Forty-eight infant rhesus macaques (30 days old) sensitized to house dust mite (HDM) were treated with two week cycles of FA, house dust mite allergen (HDMA), ozone (O{sub 3}) or HDMA/ozone (HDMA + O{sub 3}) for five months. At the end of the five months, six animals from each group were necropsied. The other six animals in each group were allowed to recover in FA for 30 more months at which time they were necropsied. Design-based stereology was used to estimate volumes of lung components, number of alveoli, size of alveoli, distribution of alveolar volumes, interalveolar capillary density. After 30 months of recovery, monkeys exposed to HDMA, in either group, had significantly more alveoli than filtered air. These alveoli also had higher capillary densities asmore » compared with FA controls. These results indicate that early life exposure to HDMA alone or HDMA + O{sub 3} alters the development process in the lung alveoli. - Highlights: • Abnormal lung development after postnatal exposure to ozone and allergen • This remodeling is shown as smaller, more numerous alveoli and narrower airways. • Allergen appears to have more of an effect than ozone during recovery. • These animals also have continued airway hyperresponsiveness (Moore et al. 2014)« less

To determine the impact of repeated episodes of ozone exposure on physiologic adaptation, epithelial injury/repair, and tracheal substance P levels, adult rats were subjected to episodes of ozone (5 days, 1 ppm, 8 h/day) followed by 9 days of filtered air for four cycles. Rats were sampled on days 1 and 5 of each episode and 9 days after day 5 of episodes 1, 2, and 4. One hour before being euthanized each rat was injected with 5-bromo-2'-deoxyuridine to label proliferating cells. Each 5-day episode showed a characteristic pattern of rapid shallow breathing (days 1 and 2), epithelial injury, and interstitial and intraluminal inflammation. In contrast, the neutrophil component of inflammation, tracheal substance P release, and cell proliferation became attenuated with each consecutive episode of exposure. Concurrent with this cyclic and attenuated response there was progressive hypercellularity and hyperplasia in all airways studied and a progressive remodeling present in the terminal bronchioles. Our findings are consistent with the notion that the cumulative distal airway lesion is at least in part the result of a depressed cell proliferative response to injury in these airways. This depressed cell proliferative response may be in part the result of diminished neutrophil inflammation and/or release of mitogenic neuropeptides in response to ozone-induced injury.

Ozone stimulates the “defensive” C-fibers in the lungs, changing breathing pattern to rapid and shallow. We hypothesized that when ozone is administered to the isolated lung with a rapid shallow breathing pattern rather than a slow deep pattern, relatively less airway epithelial damage would occur. Four groups of isolated buffer perfused rat lungs were exposed to ozone (1 ppm) or to filtered air for 90 min with either a slow deep (SDB, tidal volume 2.4 ml, frequency 40 breaths/min) or a rapid shallow breathing pattern (RSB, tidal volume 1.2 ml, frequency 80 breaths/min), resulting in an equivalent inspired dose. The absorbed dose of ozone did not differ between the exposed groups. Ethidium homodimer-1 was then instilled into the trachea to identify injured airway epithelial cells. The lungs were fixed, the airways were microdissected, and the airway epithelial cells were counterstained with YPRO-1 prior to evaluation with confocal microscopy. Ozone-induced airway epithelial cell injury occurred to a lesser overall degree when lungs were exposed by the RSB pattern (p = 0.003). The relative reduction in injury was greater (p < 0.05) in the proximal axial airway than in its adjacent airway branch and terminal bronchioles. Ozone induced an increase in pulmonary resistance with the SDB pattern but not with the RSB pattern. Thus, at an equivalent dose of inspired ozone, a RSB pattern resulted in less total damage than a SDB pattern and the distribution of protection was heterogeneous with proximal axial airways displaying the greatest relative reductions in epithelial damage.

Early morphological events associated with the genesis of epileptiform activity are essentially unknown, despite significant progress on morphological correlates of potentially related plastic neural phenomena. Hippocampal area CA1 shows the capacity to generate epileptiform bursting activity after certain patterns of electrical stimulation. Using an in vitro slice kindling preparation, we found increases in the numbers (areal densities) of shaft and sessile spine synapses in hippocampal subfield CA1 within minutes following the establishment of stable afterdischarges. These data strongly suggest that synaptogenesis is associated with the early stages of epilepsy formation.

Following induction of long-term potentiation in subfield CAl of the hippocampal slice from 26-month-old rats, shaft synapse numbers increased by 44% and sessile spine synapses (synapses on stubby, headless spines) by 72%, with the more common mushroom-shaped spine synapses statistically unaltered. These effects are smaller than in prior work with young adults, whereas population spike amplitude changes were comparable to young adults. Reasons for the discrepancy are unclear, but the results confirm continuing capacity for induction of LTP in aged rats and indicate that substantial synaptogenic capacity also persists, but may be impaired, at these ages.

The dendritic structure of neurons in the inner one-third of the wall of the barrels of somatosensory cortex was examined in animals at 10, 15, 20, 25, and 30 days of age. In adults, dendrites of these neurons project largely towards the barrel hollow. The amount of dendrite in the hemifield towards the barrel hollow, in which thalamic afferents terminate, increased with age, while the amount in the hemifield away from the hollow decreased, as measured by dendritic intersections with concentric rings. Loss of primary dendritic branches across these ages occurred both for dendrites arising on the side of the soma oriented away from the barrel hollow and for dendrites on the side of the soma nearest to the hollow. The loss of primary branches was much greater on the side away from the hollow. The amount of dendrite per primary branch increased for branches oriented towards the hollow, whereas it did not change or was somewhat reduced for those branches that remained on the side of the soma opposite the hollow. Thus both the selective loss of entire branches and the selective embellishment of others occur during the development of these somatosensory cortical structures.

A number of experimental approaches have indicated differential interneuronal connectivity following differential experience during both development and adulthood. In Golgi preparations, prolonged maze training was reported to alter dendritic branching of distal apical dendrites of Layer IV and V pyramidal neurons in adult rat occipital cortex. To determine the specificity of this effect to direct involvement in the visual aspects of training, the effects of monocular maze training, using a split-brain procedure and opaque contact occluders, was examined in the present study. Rats were maze trained with unilateral or alternating monocular occlusion, while non-trained rats with unilateral or alternating monocular occlusion were handled briefly and given water reward. There was no within-animal effect of fixed occluder position in non-trained controls. In unilaterally-occluded trained rats, Layer V pyramidal neurons in occipital cortex opposite the open eye had greater oblique dendritic length in the distal region of the apical dendrite than did those opposite the occluded eye. Similarly, rats trained with alternating occlusion had greater distal apical oblique dendritic length in Layer V occipital pyramidal neurons than did non-trained controls. This indicates that morphological sequelae of training are concentrated in areas processing information associated with visual aspects of the training and renders unlikely general metabolic or hormonal causation of such effects.

While recent studies indicate that proposed regulation of visual cortex plasticity by norepinephrine (NE) probably involves 6-hydroxydopamine (6-OHDA) effects other than NE depletion, reports of diminished neuronal maturation and reduced postweaning sensitivity to environmental conditions in animals treated with anti-adrenergic compounds continue to suggest a role for NE in neocortical development. To assess its possible role in development of a highly organized neocortical structure, the effects of postnatal 6-OHDA administration upon development of the somatosensory cortical posteromedial barrel subfield (PMBSF), which subserves the large facial whiskers, were observed in mice with whiskers in the middle row of the face removed unilaterally. Treatment with 6-OHDA caused 96–98% depletion of NE in parietal cortex. There were no effects of (or lesion interactions with) 6-OHDA treatment on barrel size in measures of Nissl-stained neurons, and 6-OHDA effects on numerical measures of dendritic branching of Golgi-impregnated non-pyramidal neurons in PMBSF were negligible. There were, however, effect of 6-OHDA treatment upon the highly ordered arrangement of dendrites within barrels. In 6-OHDA-treated animals, the class I (spiny and sparsely spined) cell dendrites were less attracted to the barrel hollow. In controls, for class I cells with their somata within the barrel wall, there was a high correlation between the distance from the soma to the hollow and the amount of dendrite in the wall, reflecting the distance the dendrite traverses to the hollow. In 6-OHDA-treated animals, this correlation was absent, i.e., cells at any distance from the hollow tended to have a high percentage of dendrite in the wall. For cells in the hollow, in controls, a relatively constant majority of dendrite stayed in the hollow. In 6-OHDA-treated animals, the amount of dendrite that hollow cells extended into the wall increased as the somata became closer to the wall, suggesting a loss of systematic governance of the orientation of dendrites by the overall pattern of the barrel. Thus the effect of 6-OHDA administration was to reduce enforcement of the normal rules of barrel organization. Two other findings related to the spread of dendrites into septa and neighboring barrels. It was found that 6-OHDA treatment increased the frequency of class II cells (smooth cells) having significant amounts of their dendrites extending outside the original barrel. Similarly, the drug treatment increased the number of class I cells extending their dendrites into adjacent denervation-induced barreloids. Given evidence that afferent activity is involved in postnatal organization of the PMBSF, all 3 findings accord with the proposal that the physiological effects of NE are to suppress background activity and to potentiate specific afferent activity, which might attract or selectively retain dendrites growing in regions of highest thalamic afferent density.

This study examined anatomical correlates of: (1) long-term potentiation (LTP); (2) equivalent low frequency synaptic activity; (3) continuous high frequency synaptic activation which did not produce LTP; and (4) synaptic inactivation by high Mg2+/low Ca2+ incubation in hippocampal subfield CA1 in the in vitro slice, and examined the persistence of changes at 10-15 min, 2 h and 8 h after stimulation. After potentiating stimulation (6 trains at 100 Hz for 1 s or 200 Hz for 0.5 s), compared to an equivalent number of low frequency stimuli (1 Hz for 600 s), there were increases in numbers of shaft and sessile spine synapses (synapses on stubby, headless spines). This suggested an increase in the number of shaft synapses onto inhibitory interneurons and/or an enhancement of synapse formation on pyramidal neurons possibly involving initial formation of shaft synapses and a transition from shaft, to sessile spine, to full grown spine synapses. Postsynaptic spine heads also assumed a rounder shape, as indicated by decreases in spine perimeter to area ratios, contact lengths, and the percentage of 'cup' shaped spines. There was no effect of potentiating stimulation on bouton or spine areas. After continuous high frequency synaptic activation (40 Hz or 100 Hz for 10 min), which produced no apparent LTP, there were no changes in synapse numbers or spine head shape parameters. However, in contrast to effects of LTP, there was an increase in bouton mitochondrial area and a marginal increase in bouton area compared to the low frequency condition. Inactivation did not affect any of these measures. LTP-associated increases in numbers of shaft and sessile spine synapses persisted over an 8 h incubation period, while the effect on spine shape disappeared after 2 h. Physiologically-demonstrable LTP persisted over the 8 h period. Effects of continuous high level activation on mitochondrial and bouton areas were even more transient, disappearing 2 h after stimulation. These findings: (1) confirm previously reported effects of potentiating stimulation on synapse numbers and spine shape; (2) indicate that spine shape changes are not necessary for the maintenance of LTP; and (3) indicate that continuous high frequency activation which does not produce potentiation has different and non-persisting effects from potentiating stimulation.