Mice and other rodents are typically utilized for chemical warfare nerve agent research. Rodents have large amounts of carboxylesterase in their blood, while humans do not. Carboxylesterase nonspecifically binds to and detoxifies nerve agent. The presence of this natural bioscavenger makes mice and other rodents poor models for studies identifying therapeutics to treat humans exposed to nerve agents. To obviate this problem, a serum carboxylesterase knockout (Es1 KO) mouse was created. In this study, Es1 KO and wild type (WT) mice were assessed for differences in gene expression, nerve agent (soman; GD) median lethal dose (MLD) values, and behavior prior to and following nerve agent exposure. No expression differences were detected between Es1 KO and WT mice in more than 34000 mouse genes tested. There was a significant difference between Es1 KO and WT mice in MLD values, as the MLD for GD-exposed WT mice was significantly higher than the MLD for GD-exposed Es1 KO mice. Behavioral assessments of Es1 KO and WT mice included an open field test, a zero maze, a Barnes maze, and a sucrose preference test (SPT). While sex differences were observed in various measures of these tests, overall, Es1 KO mice behaved similarly to WT mice. The two genotypes also showed virtually identical neuropathological changes following GD exposure. Es1 KO mice appear to have an enhanced susceptibility to GD toxicity while retaining all other behavioral and physiological responses to this nerve agent, making the Es1 KO mouse a more human-like model for nerve agent research.

Bilateral electrolytic lesions were made in the fimbria-fornix of 3-month-old male Fischer-344 rats. One week after surgery, acquisition in a shock-motivated 14-unit T-maze was assessed in these experimental animals and compared with that of sham-operated and unoperated controls. All animals were pretrained to criterion in one-way active avoidance in a straight runway before receiving two 10-trial daily sessions in the complex maze task in which the response requirement was to negotiate each of five maze segments within 10 sec to avoid footshock. All groups showed high levels of performance in pretraining. In the complex maze, rats sustaining damage to the fimbria-fornix exhibited learning impairments compared with controls in all analyzed performance measures including errors, run time, number of shocks, duration of shock, and alternation errors. The latter measure, which reflected the rat’s tendency to maintain a strategy of alternating responses at the choice-points, showed no evidence of change with training among lesioned animals, whereas control animals demonstrated a dropout of this response strategy. The impairment in maze performance associated with the fimbria-fornix lesions appeared to parallel those previously observed in scopolamine-treated young rats and in aged rats.

Calorie restriction (CR) is a reliable anti-aging intervention that attenuates the onset of a number of age-related diseases, reduces oxidative damage, and maintains function during aging. In the current study, we assessed the effects of CR and other feeding regimens on wound healing in 7-month-old Fischer-344 rats from a larger cohort of rats that had been fed either ad libitum (AL) or 40% calorie restricted based on AL consumption. Rats were assigned to one of three diet groups that received three skin punch wounds along the dorsal interscapular region (12-mm diameter near the front limbs) of the back as follows: (1) CR (n = 8) were wounded and maintained on CR until they healed, (2) AL (n = 5) were wounded and maintained on AL until wound closure was completed, and (3) CR rats were refed (RF, n = 9) AL for 48 h prior to wounding and maintained on AL until they healed. We observed that young rats on CR healed more slowly while CR rats refed for 48 h prior to wounding healed as fast as AL fed rats, similar to a study reported in aged CR and RF mice (Reed et al. 1996). Our data suggest that CR subjects, regardless of age, fail to heal well and that provision of increased nutrition to CR subjects prior to wounding enhances the healing process.

Caloric restriction (CR) mitigates neurological damage arising from aging and a variety of other sources, including neuropathology in young adult mice that express single and double transgenic (tg) mutations associated with Alzheimer disease (AD). To evaluate the potential of CR to protect against relatively heavy AD-type pathology, middle-aged (13-14-month-old) mice that co-express two mutations related to familial AD, amyloid precursor protein (APP) and presenilin 1 (PS1), were fed balanced diets with 40% fewer calories than ad libitum-fed controls. Following 18 weeks of treatment, mice were killed and brains were processed for quantification of total volume of amyloid-beta (Abeta) in the hippocampal formation and the overlying neocortex. Computerized stereology confirmed that CR reduced the total Abeta volume by about one-third compared to that in age-matched controls. Thus, CR appears to attenuate the accumulation of AD-type neuropathology in two cortical brain regions of middle-aged dtg APP/PS1 mice. These findings support the view that CR could be a potentially effective, non-pharmacology strategy for reducing relatively heavy Abeta deposition in older adult dtg APP/PS1 mice, and possibly afford similar protection against the onset and progression of AD in older adult humans.

Recent studies from our laboratories and others suggest that calorie restriction (CR) may benefit Alzheimer's disease (AD) by preventing amyloid-beta (Abeta) neuropathology in the mouse models of AD. Moreover, we found that promotion of the NAD+-dependent SIRT1 mediated deacetylase activity, a key regulator in CR extension of life span, may be a mechanism by which CR influences AD-type neuropathology. In this study we continued to explore the role of CR in AD-type brain amyloidosis in Squirrel monkeys (Saimiri sciureus). Monkeys were maintained on the normal and CR diets throughout the entire lifespan until they died of natural causes. We found that 30% CR resulted in reduced contents of Abeta1-40 and Abeta1-42 peptides in the temporal cortex of Squirrel monkeys, relative to control (CON) fed monkeys. The decreased contents of cortical Abeta peptide inversely correlated with SIRT1 protein concentrations in the same brain region; no detectable change in total full-length amyloid-beta protein precursor (AbetaPP) level was found. Most interestingly, we found that 30% CR resulted in a select elevation of alpha- but not beta- or gamma- secretase activity which coincided with decreased ROCK1 protein content in the same brain region, relative to CON group. Collectively, the study suggests that investigation of the role of CR in non-human primates may provide a valuable approach for further clarifying the role of CR in AD.

This study was designed to determine whether inhibition of formation of AGE and advanced lipoxidation end-products (ALE) is a mechanism of action common to a diverse group of therapeutic agents that limit the progress of diabetic nephropathy. We compared the effects of the ACE inhibitor enalapril, the antioxidant vitamin E, the thiol compound lipoic acid, and the AGE/ALE inhibitor pyridoxamine on the formation of AGE/ALE and protection against nephropathy in streptozotocin diabetic rats.

We examined the ability of pyridoxamine (PM), an inhibitor of formation of advanced glycation end products (AGEs) and lipoxidation end products (ALEs), to protect against diabetes-induced retinal vascular lesions. The effects of PM were compared with the antioxidants vitamin E (VE) and R-alpha-lipoic acid (LA) in streptozotocin-induced diabetic rats. Animals were given either PM (1 g/l drinking water), VE (2,000 IU/kg diet), or LA (0.05%/kg diet). After 29 weeks of diabetes, retinas were examined for pathogenic changes, alterations in extracellular matrix (ECM) gene expression, and accumulation of the immunoreactive AGE/ALE N-epsilon-(carboxymethyl)lysine (CML). Acellular capillaries were increased more than threefold, accompanied by significant upregulation of laminin immunoreactivity in the retinal microvasculature. Diabetes also increased mRNA expression for fibronectin (2-fold), collagen IV (1.6-fold), and laminin beta chain (2.6-fold) in untreated diabetic rats compared with nondiabetic rats. PM treatment protected against capillary drop-out and limited laminin protein upregulation and ECM mRNA expression and the increase in CML in the retinal vasculature. VE and LA failed to protect against retinal capillary closure and had inconsistent effects on diabetes-related upregulation of ECM mRNAs. These results indicate that the AGE/ALE inhibitor PM protected against a range of pathological changes in the diabetic retina and may be useful for treating diabetic retinopathy.

. Conditioned reflex facilitation occurs when the amplitude of the eyeblink (EB) unconditioned reflex (UR) is increased as a result of prior pavlovian conditioning. Prepulse modification of the EB reflex is produced by preceding the unconditioned stimulus (US) by a brief low-level neutral stimulus. This study examined both conditioned EB facilitation and prepulse modification in rabbits with either medial prefrontal (mPFC) lesions or sham lesions. Conditioned reflex facilitation was assessed by comparing EB UR amplitude prior to and after pavlovian EB conditioning. Animals that received CS/US paired presentations showed evidence of conditioned reflex facilitation, but animals with unpaired training did not. However, this increase in EB UR magnitude in the paired groups was smaller in animals with mPFC lesions, compared to those with sham lesions. In two subsequent experiments different groups of sham and lesion animals received an intense burst of white noise preceded by tones with different interstimulus interval durations to assess prepulse modification. Unlike conditioned facilitation, prepulse modification was unaffected by mPFC lesions.

Rabbits with lesions of either medial prefrontal cortex (mPFC) or amygdala central nucleus (ACN) were compared with sham-lesioned animals during differential and reversal classical conditioning of the eyeblink (EB) and heart rate (HR) response. Lesions of the mPFC, but not ACN, produced a severe impairment in EB reversal conditioning, but neither lesion affected original discrimination. However, both mPFC and ACN lesions produced a severe attenuation of accompanying HR decelerations during both initial differentiation and reversal. These results suggest that mPFC processing of Pavlovian conditioning contingencies affects not only the autonomic component of learning but preservative somatomotor conditioning as well, whereas ACN processing affects only the autonomic component.

Damage to the amygdala or medial prefrontal cortex (mPFC) produces deficits in learned autonomic adjustments. Whether connections between these 2 structures are necessary for acquiring classically conditioned heart rate (HR) decelerations was studied. Connections between them were interrupted by unilateral lesions of the mPFC and amygdala, but on opposite sides of the brain. One experiment focused on the mPFC and amygdala central nucleus and a second on the mPFC and amygdala basolateral complex. Bilateral lesions of either structure disrupted the HR conditioned response. The response of the 2, crossed lesioned groups also was smaller than that of the sham-operated and unilateral lesioned groups, but significant conditioned stimulus-evoked bradycardia occurred in both, suggesting that although interactions between the amygdala and mPFC may normally occur during associative emotional responding, these connections are not necessary for the acquisition of conditioned bradycardia.

Much recent evidence has shown that the thalamic-prefrontal axis is involved in Pavlovian conditioning in rabbits. However, while single cell activity in the prefrontal cortex has been previously studied during classical conditioning in rabbits, that of its thalamic projection nucleus, the mediodorsal (MD) nucleus, has not. Consequently, in the present research we recorded neuronal activity from individual cells in MD during expression of conditioned bradycardia in rabbits that received differential Pavlovian conditioning in which tones served as conditioned stimuli and periorbital shock served as unconditioned stimuli. The pattern of firing in MD was similar to that evoked in the medial prefrontal cortex (mPFC). Of 84 cells sampled, approximately 35% showed CS-evoked activity. Ninety percent of these cells showed increases in activity, while the remainder were biphasic, showing an initial increase followed by a decrease. Also, like the mPFC, some cells showed initial increases, which declined during CS presentation, while others showed gradual increases which reached their maximum at CS offset. Also some cells were responsive to the CS+ and others to the CS-. Thus, MD cells, like mPFC cells, are somewhat heterogenous with regard to responding to conditional stimuli, although, unlike the mPFC, no strictly inhibitory cells were found.

Rabbits and rats received horseradish peroxidase injections in the medial prefrontal cortex, and retrograde labeling was examined in the hippocampus (HC) and subicular complex (SC). Labeled cells were observed in HC and SC in the rat, but only in the SC of the rabbit. In a second experiment, separate groups of rabbits with sham, SC, or cortical control lesions were subjected to differential classical heart rate conditioning, in which 4-s, 75-db tones served as conditioned stimuli and a 3-mA paraorbital shock was the unconditioned stimulus. Although conditioned bradycardia was obtained in animals with SC lesions, it was slower to develop and was much shorter in duration than in the cortical and sham control groups. In the animals with SC lesions, the bradycardiac response was quickly replaced with tachycardia, suggesting a sympathetic bias in these animals.