Richard A. Bauman

Blast overpressure in rats: recreating a battlefield injury in the laboratory.

Blast injury to the brain is the predominant cause of neurotrauma in current military conflicts, and its etiology is largely undefined. Using a compression-driven shock tube to simulate blast effects, we assessed the physiological, neuropathological, and neurobehavioral consequences of airblast exposure, and also evaluated the effect of a Kevlar protective vest on acute mortality in rats and on the occurrence of traumatic brain injury (TBI) in those that survived. This approach provides survivable blast conditions under which TBI can be studied. Striking neuropathological changes were caused by both 126- and 147-kPa airblast exposures. The Kevlar vest, which encased the thorax and part of the abdomen, greatly reduced airblast mortality, and also ameliorated the widespread fiber degeneration that was prominent in brains of rats not protected by a vest during exposure to a 126-kPa airblast. This finding points to a significant contribution of the systemic effects of airblast to its brain injury pathophysiology. Airblast of this intensity also disrupted neurologic and neurobehavioral performance (e.g., beam walking and spatial navigation acquisition in the Morris water maze). When immediately followed by hemorrhagic hypotension, with MAP maintained at 30 mm Hg, airblast disrupted cardiocompensatory resilience, as reflected by reduced peak shed blood volume, time to peak shed blood volume, and time to death. These findings demonstrate that shock tube-generated airblast can cause TBI in rats, in part through systemic mediation, and that the resulting brain injury significantly impacts acute cardiovascular homeostatic mechanisms as well as neurobehavioral function.

Blast exposure in rats with body shielding is characterized primarily by diffuse axonal injury.

Blast-induced traumatic brain injury (TBI) is the signature insult in combat casualty care. Survival with neurological damage from otherwise lethal blast exposures has become possible with body armor use. We characterized the neuropathologic alterations produced by a single blast exposure in rats using a helium-driven shock tube to generate a nominal exposure of 35 pounds per square inch (PSI) (positive phase duration ∼ 4 msec). Using an IACUC-approved protocol, isoflurane-anesthetized rats were placed in a steel wedge (to shield the body) 7 feet inside the end of the tube. The left side faced the blast wave (with head-only exposure); the wedge apex focused a Mach stem onto the rats head. The insult produced ∼ 25% mortality (due to impact apnea). Surviving and sham rats were perfusion-fixed at 24 h, 72 h, or 2 weeks post-blast. Neuropathologic evaluations were performed utilizing hematoxylin and eosin, amino cupric silver, and a variety of immunohistochemical stains for amyloid precursor protein (APP), glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1 (Iba1), ED1, and rat IgG. Multifocal axonal degeneration, as evidenced by staining with amino cupric silver, was present in all blast-exposed rats at all time points. Deep cerebellar and brainstem white matter tracts were most heavily stained with amino cupric silver, with the morphologic staining patterns suggesting a process of diffuse axonal injury. Silver-stained sections revealed mild multifocal neuronal death at 24 h and 72 h. GFAP, ED1, and Iba1 staining were not prominently increased, although small numbers of reactive microglia were seen within areas of neuronal death. Increased blood-brain barrier permeability (as measured by IgG staining) was seen at 24 h and primarily affected the contralateral cortex. Axonal injury was the most prominent feature during the initial 2 weeks following blast exposure, although degeneration of other neuronal processes was also present. Strikingly, silver staining revealed otherwise undetected abnormalities, and therefore represents a recommended outcome measure in future studies of blast TBI.

Effects of chronic stress on sleep in rats

The present study was conducted to determine the effects of chronic stress on sleep using a rodent paradigm of around-the-clock signalled intermittent foot shock in which some rats can pull a chain to avoid/escape shock while another group of rats is yoked to the first group. We measured sleep using telemetry; four-channel EEG was collected 24 h/day in rats during 2 prestress days; days 1, 2, 3, 7, and 14 during chronic stress; and 3 poststress days. States of REM sleep, non-REM (NREM) sleep, and waking were scored for each 15-s period of the EEG recordings. During the prestress period, rats slept (REM plus NREM) 55% of available time during the light hours and 34% of the dark hours with the remainder represented by waking. On the first day of stress, total sleep and, especially REM sleep, decreased markedly. By the second day of stress, only REM sleep in the controllable stress group (but not the uncontrollable stress group) was still significantly decreased compared to prestress levels, and REM sleep returned to baseline levels by day 7 of stress. The recovery of sleep quantity was accomplished by increased sleep during the dark hours, resulting in a long-lasting disruption of normal circadian sleep patterning.

Effects of controllable vs. uncontrollable chronic stress on stress-responsive plasma hormones

We have previously reported effects of chronic stress on circadian rhythms of temperature, eating, and locomotor activity. These studies were conducted using an around-the-clock signalled intermittent footshock paradigm in which some rats have control over shock termination while other rats are yoked to the rats with control. Although this paradigm is stressful, as suggested by decreases in food intake and disrupted circadian rhythms, rats tolerate the paradigm well, continuing to eat, drink, gain weight, and groom. In the present studies, rats were sacrificed following 3 or 14 days of stress, and plasma was collected for hormonal assays. After 3 days of stress, plasma corticosterone and prolactin levels were elevated in both stress groups compared to controls; yoked rats had higher levels of corticosterone than rats in the group with control over shock termination, while prolactin levels in both stressed groups were similar. ACTH levels were similar in stressed and control rats. After 14 days of stress, ACTH and corticosterone levels in both stress groups were similar to control levels. Prolactin levels were elevated in the yoked experimental group compared to levels in control or controllable stress groups. These data support previous studies suggesting that control over stressors attenuates the effects of stress on physiology and demonstrate that two hormones with diverse biological effects are elevated by chronic stress.

The effect of voluntary exercise exposure on histological and neurobehavioral outcomes after ischemic brain injury in the rat.

Physical activity can induce neuroplastic adaptations and improve outcomes after cerebral injury. To determine if these outcomes are dependent on the type and timing of physical rehabilitation and the particular outcome/endpoint being tested, we evaluated the effect of voluntary exercise exposure beginning 24 h after cerebral ischemic injury on behavioral, physiological, and histological outcomes. In an observer-blinded fashion, Sprague-Dawley (300 g) male rats were allocated to three groups [sham-exercise (SHAM), stroke-exercise (SE), stroke-no exercise (SNE)] before a 1-h right middle cerebral artery occlusion (MCAo). Running wheels were used for voluntary exercise. A significant difference was found at 1 week post-infarction between the SNE and SE, with SNE showing worst neurological scores and higher number of foot faults. In addition, nearly 20% more of the SE animals regained their pre-MCAo weight by 7 days. These differences were not as evident at 2 weeks. No differences were found between the three groups in the paw preference test, wheel activity, and body temperature, as well as between SNE and SE with regards to infarct or hemispheric volumes, body weight, synaptophysin staining, and electroencephalography (EEG) testing. Within-group comparisons showed no relationships between infarct volume and foot faults, neurological scores, or exercise level. We conclude that (1) unlike behavioral outcomes, physiological and histological outcomes may not be influenced by the introduction of voluntary exercise once lesion maturation has occurred at 24 h, and (2) repetitive outcomes testing can obscure findings in rat models of cerebral ischemic injury.

Visual system degeneration induced by blast overpressure

The effect of blast overpressure on visual system pathology was studied in 14 male Sprague-Dawley rats weighing 360-432 g. Blast overpressure was simulated using a compressed-air driven shock tube, with the aim of studying a range of overpressures causing sublethal injury. Neither control (unexposed) rats nor rats exposed to 83 kiloPascals (kPa) overpressure showed evidence of visual system pathology. Neurological injury to brain visual pathways was observed in male rats surviving blast overpressure exposures of 104-110 kPa and 129-173 kPa. Optic nerve fiber degeneration was ipsilateral to the blast pressure wave. The optic chiasm contained small numbers of degenerated fibers. Optic tract fiber degeneration was present bilaterally, but was predominantly ipsilateral. Optic tract fiber degeneration was followed to nuclear groups at the level of the midbrain, midbrain-diencephalic junction, and the thalamus where degenerated fibers arborized among the neurons of: (i) the superior colliculus, (ii) pretectal region, and (iii) the lateral geniculate body. The superior colliculus contained fiber degeneration localized principally to two superficial layers (i) the stratum opticum (layer III) and (ii) stratum cinereum (layer II). The pretectal area contained degenerated fibers which were widespread in (i) the nucleus of the optic tract, (ii) olivary pretectal nucleus, (iii) anterior pretectal nucleus, and (iv) the posterior pretectal nucleus. Degenerated fibers in the lateral geniculate body were not universally distributed. They appeared to arborize among neurons of the dorsal and ventral nuclei: the ventral lateral geniculate nucleus (parvocellular and magnocellular parts); and the dorsal lateral geniculate nucleus. The axonopathy observed in the central visual pathways and nuclei of the rat brain are consistent with the presence of blast overpressure induced injury to the retina. The orbital cavities of the human skull contain frontally-directed eyeballs for binocular vision. Humans looking directly into an oncoming blast wave place both eyes at risk. With bilateral visual system injury, neurological deficits may include loss or impairments of ocular movements, and of the pupillary and accommodation reflexes, retinal hemorrhages, scotomas, and general blindness. These findings suggest that the retina should be investigated for the presence of traumatic or ischemic cellular injury, hemorrhages, scotomas, and retinal detachment.

Effects of controllable vs. uncontrollable stress on circadian temperature rhythms

The effects of sustained stress on body temperature were investigated in rats implanted with mini-transmitters that permitted remote measurement of body temperature. Temperature was first monitored during control conditions. Following the control period, rats were either shaped to avoid/escape signalled around-the-clock intermittent footshock (controllable stress) or yoked to the controlling rats such that the controlling rat and the yoked rat received shock of the same duration, but only the controlling rat could terminate shock by pulling a ceiling chain. Under control conditions, rats demonstrated regular rhythms in body temperature which averaged 1 degree higher during the 12-h dark cycle than the light cycle. Stress disrupted the rhythm and markedly decreased the night-day difference in temperature, especially in the yoked rats in which almost no difference between light and dark cycle temperature was seen. The disruption was most marked for the first days of stress. A regular temperature rhythm was reestablished following about 5 days of stress although the stress condition continued. Leverpressing for food was also affected by the stress conditions with both stress groups leverpressing less than controls and the uncontrollable stress group pressing less than the controllable stress group. These data offer additional evidence of the increased pathophysiological effects of uncontrollable as compared to controllable stress.

Rapid demand curves for behavioral economics

A method for determining the relationship between food consumption and the price of food (demand function) in behavioral economic experiments is described. Although previous methods have generally required as long as 40 days, the present method can generate a complete demand function within 7 days, and therefore may be more suitable for use in the evaluation of drugs, toxins, and physiological/anatomical interventions. Moreover, measures of circadian rhythmicity, post-reinforcement pause durations, and interresponse times can also be generated. Three experiments tested the stability of the method in a variety of procedural manipulations: repeated exposure to the procedure, increasing versus random daily food price, and size of daily changes in food price. The procedure generated demand functions that were similar to those that require more extended testing, and the demand functions were not generally affected by procedural manipulations. Body weight, which can also affect consumption, generally decreases with increases in the price of food; so this variable should be recorded and used as a covariate in analyzing demand functions. With the exception of circadian rhythmicity, the other measures were stable across procedural variations and showed expected changes as a function of food price: postreinforcement pause durations increased as price increased, but interresponse times did not.

Effects of chronic stress on food acquisition, plasma hormones, and the estrous cycle of female rats

Our laboratory has previously conducted a number of studies to determine the effects of chronic stress on the physiology and behavior of male rats. The present study was performed to extend these investigations to female rats. Female rats were chronically stressed using a behavioral paradigm of around-the-clock signalled intermittent foot shock in which some rats can pull a chain to avoid/escape shock (stress) while another group of rats is yoked to the first group (yoked-stress) and does not have control over shock termination. Control rats were never shocked but all groups lever pressed for food pellets on an FR1 schedule (one pellet per lever press). Daily vaginal samples were obtained for several weeks prior to stress onset and throughout the chronic stress period. After 14 days of stress, the experiment was terminated and morning blood samples were collected for hormonal assays. Stress transiently decreased lever pressing for food pellets and body weights, but both measures returned to prestress levels by day 14 of stress. Plasma adrenocorticotropic hormone (ACTH) concentrations were significantly elevated in the yoked-stress group compared to the other two groups, but there were no significant effects of 14 days of stress treatment on plasma corticosterone, prolactin, estradiol, or progesterone concentrations. There were no significant differences in estrous cycle length among experimental groups.

The Quantitative Analysis of Economic Behavior With Laboratory Animals

Studies of animal behavior in the laboratory provide an extensive data set on economic behavior under controlled and replicable conditions. In the studies reported, the food consumption of rodents and non-human primates was studied under a wide variety of conditions of work effort, commodity value, and substitute availability. The results conformed to a single underlying demand function that assumed that demand elasticity was linear in price. Price was defined as a cost-benefit ratio that included consideration of both the effort of the work and the value of the commodity purchased. The parameters of the elasticity equation appeared related to the amount and temporal proximity of a substitute food.