Traumatized Rats Research Articles

Sensitivity to trauma-associated cues is restricted to vulnerable traumatized rats and reinstated after extinction by yohimbine

While post-traumatic stress disorder (PTSD) symptom is mainly characterized by re-experiencing the traumatic event, the reactivity to trauma-associated cues in resilient and vulnerable subjects has not been extensively studied. Using an animal model of PTSD induced by a single prolonged stress (SPS), the responses of traumatized Vulnerable and Resilient rats to PTSD-like symptom tests and to trauma-associated cues were investigated. In addition, the implication of the noradrenergic system in “re-experiencing” was explored. Rats received either a SPS, combining a 2h restraint stress, a 20min forced-swim followed by a 15min rest, and a loss of consciousness produced by inhaling CO2 emissions, delivered in the presence of particular cues (tone and odor), or a control procedure. PTSD-like symptoms and reactivity to various trauma-associated cues (specific, contextual, or predictive) were tested from D15 to D60 after the SPS. Rats were then divided into Resilient and Vulnerable on the basis of three main symptom tests, including the elevated plus maze, the light-dark and the acoustic startle response tests. Although Resilient rats behaved like Controls rats, Vulnerable rats developed long-term PTSD-like symptoms on the main symptoms tests (anxiety and alteration of arousal), as well as other PTSD-like outcomes (such as anhedonia and avoidance to trauma-associated cues). These Vulnerable rats were also the only ones to demonstrate strong reactivity to trauma-associated cues. In addition, the alpha-2 adrenergic receptor antagonist, Yohimbine (i.p., 1.5mg/kg/ml), was able to reinstate fear responses to an extinguished trauma-associated odor. Our results established clear relationships between Vulnerability to trauma and reactivity to trauma-associated cues and further suggest an involvement of the noradrenergic system.

Silicon Dioxide Nanoparticles (SiO2, 40–50 nm) Exacerbate Pathophysiology of Traumatic Spinal Cord Injury and Deteriorate Functional Outcome in the Rat. An Experimental Study Using Pharmacological and Morphological Approaches

Silicon (SiO2) nanoparticles or silica dust is quite common form of exposure to soldiers engaged in gulf war that may influence their health and brain function. It is quite likely that traumatic injuries to the CNS may be influenced by exposure to these nanoparticles. However, the details of silicon nanoparticles on human health functions are still unknown. In this investigation we examined the effects of chronic silicon dioxide nanoparticles (SiO2, 40-50 nm) exposure on spinal cord injury (SCI) induced alterations on the functional outcome and the cord pathology in a rat model. Since nanoparticles induce oxidative stress, the influence of an antioxidant compound H-290/51 was also examined in these nanoparticles treated injured rats as well. Rats treated with SiO2 for 7 days did not show any significant alteration in behaviour on rota rod performances or on capacity angle tests. However, subjection of these nanoparticles exposed rats to SCI resulted in a profound deterioration in motor functions compared to normal rats with SCI. The magnitude of blood-spinal cord barrier (BSCB) disruption to Evans blue and radioiodine tracers and edema formation was much more aggravated following SCI in nanoparticles treated animals compared to untreated traumatized rats. Pretreatment with H-290/51 (50 mg/kg, p.o.) 30 min before SCI in nanoparticle treated rats did not alter spinal cord pathology or functional outcome, however, this dose of the compound was very effective in reducing pathophysiology of SCI in normal animals. These observations are the first to suggest that exposure of nanoparticles enhances the sensitivity of CNS to injuries and alter the effect of neuroprotective drugs.

Rats exposed to traumatic stress bury unfamiliar objects — A novel measure of hyper-vigilance in PTSD models?

Electric shocks lead to lasting behavioral deficits in rodents, and as such are often used to model post-traumatic stress disorder (PTSD) in the laboratory. Here we show that a single exposure of rats to 3 mA-strong shocks results in a marked social avoidance that lasts at least 28 days; moreover, the response intensifies over time. In an attempt to study the impact of cue reminders on the behavior of shocked rats, we administered shocks in the presence of a highly conspicuous, 10 cm-large object. This object was introduced into the home cage of rats 28 days after shock exposure. Shocked rats manipulated the object considerably less than controls. More importantly, however, the object was buried by shocked rats. This behavior was virtually absent in controls. The response strongly depended on the intensity of shocks, and was robust. Rats shocked with 3 mA currents spent 40% of time burying the object, which was often hardly visible at the end of the 5 min test. Subsequent experiments demonstrated that the response was not cue-specific as unfamiliar objects were also buried. Rats are well known to bury dangerous objects; the shock-prod burying test of anxiety is based on this response. Behavioral similarities with this test and the differences from the marble-burying behavior of mice suggest that traumatized rats bury unfamiliar objects in defense, and the response can be interpreted as a sign of hyper-vigilance. We further suggest that object burying can be used as a sign of hyper-vigilance in models of PTSD.

Influence of posttraumatic hypoxia on behavioral recovery and histopathological outcome following moderate spinal cord injury in rats.

Pulmonary dysfunction leading to secondary hypoxia is a common complication of spinal cord injury (SCI). The purpose of this study was to clarify the behavioral and histopathological consequences of posttraumatic hypoxia in an established model of traumatic SCI. Forty-five female Sprague-Dawley rats were randomly assigned to one of four groups, including (1) laminectomy and normoxia (n = 10), (2) laminectomy and hypoxia (n = 11), (3) NYU weight-drop and normoxia (n = 12), and (4) NYU weight-drop and hypoxia (n = 11). For these studies, a moderate injury was induced by adjusting the height of the weight drop (10 g) to 12.5 mm above the exposed spinal cord (T10). Immediately after injury, PaO2 in the hypoxic rats was kept between 30 and 35 mm Hg for 30 min. PaO2 in the normoxic group was maintained over 100 mm Hg, while PaCO2 in all rats was maintained at 35-40 mm Hg. The behavior of the rats was checked every 7 days using the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale. Rats were sacrificed at 8 weeks for quantitative histopathological analysis of lesion areas. During the hypoxic insults, the mean arterial blood pressure dropped in both sham control and weight-drop rats (p < 0.01). At the end of the 8-week monitoring period, BBB scores were 12.5 +/- 3.1 (mean +/- SEM) and 14.2 +/- 3.4 in the normoxic and hypoxic traumatized rats, respectively. No significant difference between the traumatized groups was documented with BBB monitoring. In contrast, the percent of gray matter necrosis at the impact epicenter was significantly increased in hypoxic versus normoxic SCI rats (p < 0.01). These data demonstrate that posttraumatic hypoxia complicated by mild hypotension aggravates the histopathological consequences of SCI and further emphasize the need to control for secondary hypoxic insults after experimental and clinical SCI. Potential explanations for the lack of a correlation between the behavioral and histopathological findings are discussed.

Upregulation of Uncoupling Protein Homologues in Skeletal Muscle but Not Adipose Tissue in Posttraumatic Insulin Resistance

Metabolic alterations after surgical stress include peripheral insulin resistance and increased utilization of fat as a fuel substrate. An up-regulation of skeletal muscle uncoupling proteins (UCPs) has been associated with physiologic states of insulin resistance and enhanced fat metabolism in rodents. We examined whether posttraumatic insulin resistance induced the UCPs in gastrocnemius and soleus muscle and white adipose tissue in an experimental model of surgical trauma. Insulin sensitivity was significantly reduced in isolated soleus muscles but unchanged in adipocytes after trauma. In traumatized rats, mRNA and protein contents of UCP2 and UCP3 and were significantly increased in both muscle types. UCP2 protein content in adipose tissue was unaltered by surgical stress. Circulating NEFAs and glycerol were reduced after surgical trauma. We hypothesize that the changes in UCP2 and UCP3 gene and protein expression are involved in the regulation of substrate utilization in posttraumatic insulin resistance.

Changes of biological reducing activity in rat brain following closed head injury: a cyclic voltammetry study in normal and heat-acclimated rats.

Reactive oxygen species (ROS) are normally generated in the brain during metabolism, and their production is enhanced by various insults. Low molecular weight antioxidants (LMWA) are one of the defense mechanisms of the living cell against ROS. The reducing capacity of brain tissue (total LMWA) was measured by cyclic voltammetry (CV), which records biological oxidation potential specific to the type of scavenger(s) present and anodic current intensity (Ia), which depends on scavenger concentration. In the present study, the reducing capacity of rat brain following closed head injury (CHI) was measured. In addition, CV of heat-acclimated traumatized rats was used to correlate endogenous cerebroprotection after CHI with LMWA activity. Sham-injured rat brains displayed two anodic potentials: at 350 +/- 50 mV (Ia = 0.75 +/- 0.06 microA/mg protein) and at 750 +/- 50 mV (Ia = 1.00 +/- 0.05 microA/mg protein). Following CHI, the anodic waves appeared at the same potentials as in the sham animals. However, within 5 min of CHI, the total reducing capacity was transiently decreased by 40% (p < 0.01). A second dip was detected at 24 h (60%, p < 0.005). By 48 h and at 7 days, the Ia levels normalized. The acclimated rats displayed anodic potentials identical to those of normothermic rats. However, the Ia of both potentials was lower (60% of control, p < 0.001). The Ia profile after CHI was the direct opposite of the normothermic Ia profile: no immediate decrease of Ia and an increase from 4 h and up to 7 days (40-50%, p < 0.001). We suggest that the lowered levels of LMWA in the post-CHI period reflect their consumption due to overproduction of free radicals. The augmented concentration of LMWA found in the brain of the heat-acclimated rats suggests that these rats are better able to cope with these harmful radicals, resulting in a more favorable outcome following CHI.

Antihistaminergic pretreatment prevents tissue extravasation of albumin from intra-abdominal trauma in rats.

Intra-abdominal surgery causes a loss of plasma into tissues within and around the abdomen, endangering tissue viability. Mast cells, containing histamine, are abundant in the abdominal cavity. In a model of mechanical intra-abdominal trauma, we investigated whether pretreatment with histaminergic H1 and H2 receptor blockers counteracts this extravasation. In Wistar rats under chloralose anaesthesia, tissue clearances of labelled albumin were determined by a double isotope technique. Four groups were studied: Traumatized rats, pretreated (n = 10) and non-pretreated (n = 9): non-traumatized rats; pretreated (n = 10) and non-pretreated (n = 9). In the traumatized rats, given pyrilamine 10 mg kg-1 (H1) plus cimetidine 25 mg kg-1 (H2) just before the trauma, the blockers prevented the haemoconcentration from loss of plasma and the drop in arterial pressure during the very trauma procedure, observed among non-pretreated rats. Furthermore, they significantly decreased the clearance of albumin in the abdominal wall and the pancreas. In the non-traumatized animals, the blockers lowered arterial pressure and heart rate. In conclusion, the anti-histaminergic pretreatment decreased the trauma induced leakage of albumin, by mechanisms which may involve readjustments of pressures and flows in capillaries as well as a prevention of histamine effects on capillary permeability.

Early microvascular and neuronal consequences of traumatic brain injury: a light and electron microscopic study in rats.

The purpose of this study was to document the early morphologic consequences of moderate traumatic brain injury (TBI) in anesthetized Sprague-Dawley rats. Normothermic rats (37 degrees C) were injured with a fluid percussion pulse (1.7-2.1 atm) administered by an injury cannula positioned parasagittally over the right cerebral cortex (n = 7). At 45 min following TBI, rats were injected with the protein tracer horseradish peroxidase (HRP) and perfusion fixed or immersion fixed 15 min later for light and electron microscopic analysis. Blood-brain barrier (BBB) breakdown to HRP was present overlying the pial surface and superficial cortical layers of the injured hemisphere. A focal area of severe HRP leakage was also present at the gray-white interface of the lateral cortex. Light microscopic examination of this site revealed petechial hemorrhages associated with small venules. Dark shrunken neurons and swollen astrocytes were detected within cortical areas overlying the evolving contusion, CA3 and CA4 hippocampal subsectors, and lateral thalamus. Ultrastructural studies obtained evidence for irreversible neuronal injury and mechanical damage to vessel walls at this early posttraumatic period. In nonperfused traumatized rats, luminal platelet aggregates were also detected at sites of hemorrhage. In this model of TBI, a consistent pattern of microvascular and neuronal abnormalities can be documented in the early posttraumatic period. Pathomechanisms underlying these early changes are discussed in terms of primary and secondary injury processes.

Altered tissue polyamine levels due to ornithine-α-ketoglutarate in traumatized growing rats

All cells contain significant amounts of polyamines (PA), and their concentrations are highly regulated. Metabolic activity within a tissue may be reflected in the amount of intracellular PA. Since trauma involves accelerated death and regeneration of tissues, the related levels of PA in extracellular and intracellular fluids may reflect altered protein metabolism. Trauma induces an increased excretion of urinary PA, and the tissues responsible for this whole-body activity are not known. During posttraumatic nutritional management, supplementation with ornithine-α-ketoglutarate (OKG) seems to improve nitrogen economy. The present study evaluates the significance of muscle, liver, and intestine PA responses in traumatized (bilateral femur fractures) rats to the feeding of an isonitrogenous liquid diet supplemented with or without OKG. Uninjured control rats were pair-fed with respective traumatized rats. After 2 days of starvation and 4 days of feeding, the traumatized and control rats were killed and the tissues were excised and analyzed. Starvation decreases and refeeding increases urinary PA excretion. Trauma-induced PA response is predominantly seen in muscle tissues, and this may be responsible for parallel increases in PA excretion. Liver PA responses show a varying tendency confirming the increased protein synthetic activity due to trauma. Intestine has the highest intracellular PA levels, and there is a general smaller (statistically insignificant) increase in all the individual PA contents due to trauma. OKG supplementation augments tissue and urine PA responses in control rats; however, in trauma rats muscle PA levels show very little change, although nitrogen retention is significantly better (88% to 77%). Mechanistic studies are needed to evaluate the significances of the time-dependent, injury-induced, individual intracellular PA levels.

Nitrogen retention in rats fed on diets enriched with arginine and glycine. 1. Improved N retention after trauma.

1. Nitrogen retention was measured in adult rats (250-350 g) subjected to the trauma of hind-leg fracture and given diets with or without arginine plus glycine supplementation. Observations were also recorded on excretion of creatine, creatinine, allantoin, and orotic acid. Liver and skeletal muscle transaminase activities were also determined. 2. When traumatized rats weighing approximately 250 g were given a diet with 200 g casein/kg, supplemented with 20 g arginine and 10 g glycine/kg (EC diet) or a casein diet made isonitrogenous with the EC diet by addition of aspartic acid (C diet), a 60-70% increase in N retention was observed for the first 5 d post-injury for animals consuming the EC diet. A soya-bean (S) diet, isonitrogenous to the diet containing 20% casein, supplemented with arginine and glycine was as effective as the EC diet in promoting significantly better N retention of traumatized rats (350 g) in comparison to rats given the C diet. 3. When the dietary casein content was reduced to 100 g/kg, supplements of 10 g arginine and 5 g glycine or 20 g arginine and 10 g glycine/kg did not improve N retention. It is suggested that both protein quality and protein quantity are important following injury. 4. An increased excretion of creatine was observed in traumatized rats given the high-protein diets supplemented with arginine and glycine. No consistent changes were noted for urine creatinine. 5. 5. Urine allantoin levels remained stable after leg-fracture in rats consuming either the C or EC diets. Differences in the levels of urine orotic acid were found during both the pre- and post-injury periods in rats given the C, EC or S diets. 6. The mechanisms responsible for the improved N retention of traumatized rats consuming the high-protein diets with supplements of arginine and glycine may be related to the role of arginine both as a constituent of muscle tissue and as an intermediate in the urea cycle. 7. In traumatized rats fed the C or EC diets, liver transaminase activity increased whereas the transaminase activity in skeletal muscle decreased. These results support the recent concept that the increased excretion of N following injury arises from diminished reutilization of amino acids by muscle tissue without an acute increase in the rate of muscle catabolism.

Effect of high lipid diets on normal and traumatized rats.

Rats were maintained on 40% butter, 40% corn oil and control diets for 2, 4 and 6 weeks. Rats from each diet were traumatized in the Noble-Collip Drum so as to compare plasminogen activation. Serum proteolytic activity and cholesterol levels were determined. Differences in weights of the comparably dieted animals were negligible. Serum cholesterol levels were markedly higher in those fed high fat diets with 5% cholesterol than in rats on control diets. Trauma caused significantly increased serum proteolytic activity and serum cholesterol levels in all groups. The diet administered produced moderate hypercholesterolemia which did not significantly alter the serum proteolytic activator or plasmin activity of normal or traumatized rats.

Metabolism following a turpentine abscess in old age

Changes in the level of free fatty acids in serum, aorta, and adipose tissue were found to be more pronounced in young than in old female rats traumatized with a turpentine abscess on their hind paw. A similar difference was found for the level of urea in the blood. The blood level of pyruvate was increased in young rats only. The degree of inflammation was the same in both age groups. It is concluded that young animals mobilize more fat, proteins, and carbohydrates after trauma than old ones. The elevation of serum mucoproteins was the same in old and young traumatized rats.

FACTORS DETERMINING CESSATION OF CORPUS LUTEUM FUNCTION; THE POSSIBLE ROLE OF OESTRADIOL AND PROGESTERONE.

ABSTRACT The role of oestrogen and progesterone utilization by the uterus was investigated with regard to the mechanism of cessation of corpus luteum function. After the administration of oestradiol benzoate with or without progesterone in various combinations to spayed, spayed-hysterectomized, or spayed traumatized rats it was found that: there were no percentage differences in the oestrogenic effects in the vaginal smears of spayed or spayed-hysterectomized rats after the administration of oestradiol benzoate alone. After treatment with combinations of oestradiol and progesterone, the oestrogenic effects were inhibited in spayed-hysterectomized animals as compared with similarly treated spayed controls. A still more marked inhibition was obtained after traumatizing the uterine endometrium. Increasing doses of progesterone in combination with a fixed dose of oestradiol benzoate progressively delayed vaginal opening. It is concluded that the uterus does not utilize any measurable amounts of oestrogen, but that, on the other hand, it does utilize considerable quantities of progesterone. Traumatization of the uterus may have a similar progesterone-sparing effect. The findings are discussed against the background of factors which determine cessation of corpus luteum function, while it is suggested that progesterone may be an important factor accounting for the effects of hysterectomy.