Conditioning In Rabbits Research Articles

Influence of baroreflex on volume elasticity of heart and aorta in the rabbit.

Optimal ventriculoaortic coupling includes tuning of elastic properties. The ratio of effective arterial elastance and left ventricular endsystolic elastance is often taken as a measure for mechanical and energetical efficiency. The present study determined the time course of ventricular and aortic volume elasticity (VE = dp/dV) throughout a complete heartbeat. This was achieved by using changes of eigenfrequency of two catheter-transducer systems under closed chest conditions in rabbits. Short-term VE modulation was studied by a baroreflex response, as induced by pressure changes applied to the carotid sinus. Long-term changes were studied in atherosclerotic rabbits (12 wk of high-cholesterol feeding). The time course and mean values of ventricular and aortic VE were changed by the baroreflex stimulus. Cholesterol feeding diminished the response. The degree of ventriculoaortic coupling, as quantified by VE(Aorta)/VE(Ventricle) ratio, varied during a single ejection period. The large span allows either maximal energetical efficiency or maximal stroke work. Although normal rabbits adjusted their ventriculoaortic coupling during baroreflex input, the cholesterol-fed rabbits failed to do so.

Timing at the Start of Associative Learning

Some theories of associative learning imply that time plays a fundamental role in the acquisition process. Consistent with these theories, this paper presents evidence that the time from the onset of a conditioned stimulus (CS) until presentation of the unconditioned stimulus (US) is learned very rapidly at the start of training. We report two autoshaping studies and a study on aversive conditioning in goldfish in which we examine timing at the start of conditioning. We also review data from a number of other conditioning preparations, including fear-potentiated startle, appetitive conditioning in rats, and eyeblink conditioning in rabbits, that report conditioned response (CR) timing early in training. Acquisition speed and the very first expressions of conditioned responding often show sensitivity to the time of US presentation. In instances where temporal control is slowly expressed, it is likely due to performance factors, not to slow learning about time. In fact, the learning about time may be a necessary condition for associative learning.

Intra-cerebellar infusion of NMDA receptor antagonist AP5 disrupts classical eyeblink conditioning in rabbits

Rabbits were infused with AP5, an NMDA receptor antagonist, into the region of the cerebellar interpositus nucleus during classical eyeblink conditioning with a tone conditioned stimulus and an air puff unconditioned stimulus. Acquisition of the conditioned eyeblink response was delayed in rabbits infused with AP5 but the NMDA receptor antagonist had little effect on conditioned responses when these same rabbits were infused a second time after reaching asymptotic responding levels. Some rabbits that received AP5 infusions for the first time after the conditioned response was well learned showed temporary alterations in response timing. These data indicate that NMDA receptor activity is involved in the acquisition of classically conditioned eyeblink response and may also be involved in regulating cellular processes involved in response timing and other aspects of conditioned response execution.

Effects of estrogen on cerebral blood flow and pial microvasculature in rabbits.

We tested the hypothesis that intracarotid estrogen infusion increases cerebral blood flow (CBF) in a concentration-dependent manner and direct application of estrogen on pial arterioles yields estrogen receptor-mediated vasodilation. Rabbits of both genders were infused with estrogen via a branch of the carotid artery. Estrogen doses of 20 or 0.05 microg. ml(-1). min(-1) were used to achieve supraphysiological or physiological plasma estrogen levels, respectively. CBF and cerebral vascular resistance were determined at baseline, during the infusion, and 60-min postinfusion, and effects on pial diameter were assessed via a cranial window. Pial arteriolar response to estrogen alone and to estrogen after administration of tamoxifen (10(-7)), an antiestrogen drug that binds to both known estrogen receptor subtypes, was tested. No gender differences were observed; therefore, data were combined for both males and females. Systemic estrogen infusion did not increase regional CBF. Estradiol dilated pial arteries only at concentrations ranging from 10(-4)-10(-7) M (P < or = 0.05). Pretreatment with tamoxifen alone had no effect on arteriolar diameter but inhibited estrogen-induced vasodilation (P < 0.001). Our data suggest that estrogen does not increase CBF under steady-state conditions in rabbits. In the pial circulation, topically applied estradiol at micromolar concentrations dilates vessels. The onset is rapid and dependent on estrogen receptor activation.

Microinfusion of protein kinase inhibitor H7 into the cerebellum impairs the acquisition but not the retention of classical eyeblink conditioning in rabbits

Rabbits were infused with H7, a general protein kinase inhibitor, into the region of the cerebellar interpositus nucleus during classical eyeblink conditioning. Acquisition of the conditioned eyeblink response was delayed by the H7 infusion, but the protein kinase inhibitor had no effect on performance of the learned response when infused after asymptotic learning had been reached. These data indicate that protein kinases in the cerebellum are involved in plasticity processes that underlie the learning of this simple conditioned behavior.

Neurotoxic lesions of the dorsal hippocampus disrupt auditory-cued trace heart rate (fear) conditioning in rabbits.

The first experiment in this study used the classical heart rate (HR) conditioning paradigm to determine if rabbits could associate an auditory conditioned stimulus (CS) and a fear-producing shock-unconditioned stimulus (US) separated by an empty 10-s trace interval. Trace conditioned rabbits (n = 7) acquired significant bradycardiac conditioned HR responses on CS-alone test trials during a single 35-trial conditioning session. Control animals (n = 7) which received unpaired CSs and USs did not show HR conditioning. During a retention session of CS-alone trials 24 h after the conditioning session, some trace-conditioned animals showed conditioned HR responses immediately following CS onset (n = 3), while others showed responses appropriately timed to the US onset (n = 4) used in trace conditioning 24 h earlier. Thus, rabbits remember the duration of the long 10-s trace interval 24 h after a single day of training. The second part of this study sought to determine if cells in the dorsal hippocampus play a role in trace HR conditioning. Rabbits were given bilateral ibotenic acid lesions in the neocortex (n = 7) or dorsal hippocampus (n = 8). During trace conditioning and retention, neocortical animals showed conditioned HR responses to the CS, whereas the hippocampal group showed no significant HR conditioning. One week after trace conditioning, the same animals received a delay HR conditioning session where no trace interval separated the CS and US. During delay conditioning, hippocampal animals showed significant conditioned HR responses to the CS that were similar to the neocortical group. Thus, the dorsal hippocampus plays a critical role in rabbit HR conditioning when the CS and US are separated by a 10-s trace interval. This paradigm may be ideal for in vivo electrophysiological recording studies because rabbits are easily immobilized during the testing procedure, and learning occurs during a single day of training.

Dilution is effective in reducing infusion phlebitis in peripheral parenteral nutrition: An experimental study in rabbits

To clarify conflicting clinical results that had been reported as to whether dilution is effective or not in reducing infusion phlebitis, this study was undertaken. We undertook two experiments with the different infusion conditions in rabbits to confirm the generality and the reproducibility of the results. To test the effect of dilution, 120 mL/kg of solution A (784 mOsm/kg) was infused into rabbit ear veins at 10 mL.kg-1.h-1 for 12 h, and 144 mL/kg of 1.2-fold-diluted solution A (648 mOsm/kg) was infused at 12 mL.kg-1.h-1 for 12 h. Similarly, 120 mL/kg of solution B (718 mOsm/kg) was infused at 5 mL.kg-1.h-1 for 24 h, and 168 mL/kg of 1.4-fold-diluted solution B (514 mOsm/kg) was infused at 7 mL.kg-1.h-1 for 24 h. The infused veins were sampled 24 h after the end of the infusion and examined histopathologically. After the 12-h infusion, phlebitic changes were observed in six of eight rabbits given solution A but in only one of eight rabbits given diluted solution A, although the same quantities of the same nutrients were infused. Also, after the 24-h infusion, phlebitic changes were observed in six of eight rabbits given solution B but in no animals given diluted solution B. The same result that dilution reduced or eliminated phlebitic changes was confirmed in the different conditions. These results suggest that osmolality of the infusion solution is an important factor in the development of phlebitis regardless of infusion volume or infusion rate and that dilution is effective in reducing the phlebitic potential of infusion solutions.

Characterization of Single-Unit Activity in the Mediodorsal Nucleus of the Thalamus during Expression of Differential Heart Rate Conditioning in the Rabbit

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.

Hippocampus in delay eyeblink classical conditioning: essential for nefiracetam amelioration of learning in older rabbits

Rabbits acquire conditioned responses (CRs) normally with bilateral removal of the hippocampus, but alterations of the intact hippocampus can affect the rate of acquisition. The cognition-enhancing drug, nefiracetam ameliorated the acquisition of CRs in older rabbits, protected membrane dysfunction in hippocampal CA1 neurons following oxygen and glucose deprivation, and promoted the release of diverse neurotransmitters, including acetylcholine. Because the septo-hippocampal cholinergic system is demonstrated to be involved in eyeblink conditioning, this experiment was undertaken to explore whether nefiracetam ameliorates conditioning via the hippocampus. Data from 53 rabbits of a mean age of 28 months were tested under two drug conditions (10 or 0 mg/kg nefiracetam) and 4 lesion conditions (bilateral hippocampectomy, bilateral neocortical removal, sham surgery, no surgery). The three groups of nefiracetam-treated rabbits with intact hippocampus acquired CRs more rapidly than the vehicle-treated groups, but rabbits with bilateral hippocampectomy treated with nefiracetam learned like vehicle-treated rabbits. Results suggest that nefiracetam ameliorates learning via the hippocampus. Because of the parallels between conditioning in rabbits with disrupted hippocampal cholinergic systems and conditioning in Alzheimer's disease (AD), these results suggest that nefiracetam may ameliorate conditioning in AD as it ameliorates conditioning in older rabbits.

Hippocampal activity during classical discrimination--reversal eyeblink conditioning in rabbits.

Multiple-unit neuronal recordings were taken from the hippocampi of 10 male, New Zealand white rabbits during classical discrimination and reversal eyeblink conditioning using 2 tones as the conditioned stimuli (CS+ and CS-) and an air-puff unconditioned stimulus. During discrimination training, characteristic learning-related activity was seen in the hippocampus on trials when a conditioned response (CR) was executed. During early phases of reversal training, however, when high numbers of CRs were evident to both the new CS+ (the former CS-) and the new CS- (the former CS+), no learning-related activity was observed. Characteristic CR-related hippocampal activity to the CS+ was observed only after the rabbits began to learn the reversal response. These results suggest that the hippocampus may encode different features of eyeblink conditioning during discrimination and reversal learning.

Hippocampal activity during classical discrimination—Reversal eyeblink conditioning in rabbits.

Hippocampal activity during classical discrimination—Reversal eyeblink conditioning in rabbits.

The effect of Dotarizine (Ca2+ channel blocker) on cerebral vessel reactivity in animals subjected to hyperventilation and anoxia.

Dotarizine--a novel piperazine derivative--belongs to wide spectrum Ca+ channel antagonists. It was reported to have strong vasodilatory and antiserotoninergic activities. Unlike other Ca+ channel blockers Dotarizine was found to have lower oral toxicity. In the presented study the influence of the oral administration of the novel compound on the blood flow velocity changes in different cerebral arteries--in basilar artery (BA) and middle cerebral artery (MCA)--was investigated under hyperventilation and hypoxic conditions of rabbits. In the first experimental group 25 mg/kg of Dotarizine dissolved in 0.25% agar was administered orally three times at the 10 hours' intervals. The sham group of animals was fed with agar of the same concentration. The results revealed that oral administration of Dotarizine diminished the vasoconstrictive effect of hyperventilation and this was more pronounced in MCA than in BA. During anoxic conditions stronger vasodilatory effects were observed in both groups of vessels and the low value of pulsatility index (PI) reflected pronounced decrease of the peripheral resistance, in comparison to the control group. Thus, the oral administration of Dotarizine decreases the peripheral resistance of cerebral vessels and therefore seems to have influence on the minute arteries of cerebrovascular system of the brain.

Neural substrates of discriminative avoidance learning and classical eyeblink conditioning in rabbits: a double dissociation

In a previous study, lesions of the deep cerebellar nuclei blocked classical eyeblink conditioning, but did not impair discriminative avoidance learning in rabbits. Here, as also found previously, lesions of the anterior and medial dorsal thalamic nuclei severely impaired discriminative avoidance learning. However, these lesions had no impact on discriminative eyeblink conditioning or reversal learning. These results complete the demonstration of a double dissociation, indicating distinct neural substrates for the acquisition of these learned behaviors. It is proposed that the two learning circuits identified by these studies mediate, respectively, acquisition of specific adaptive reflexes and whole-body, voluntary goal-directed movements.

Trace Eyeblink Conditioning in Rabbits Demonstrates Heterogeneity of Learning Ability Both Between and Within Age Groups

Rabbits 2 to 41 months of age were conditioned in the 500 ms trace eyeblink paradigm to cross-sectionally define the age of onset and the severity of age-associated impairments in acquisition of this relatively difficult hippocampally dependent task. Using a strict behavioral criterion of 80% conditioned responses (CRs), age-associated learning impairments were significant by 24 months of age. Among rabbits that successfully reached this criterion, impairments in acquisition plateaued at 30 months of age. However, the proportion of severely impaired rabbits (that failed to reach the 80% criterion) continued to increase age dependently. Using an easier criterion of 8 out of 10 CRs, behavioral impairments were not detected until 30 months of age, and cases of severe impairment (failure to reach criterion) were rare. Additional controls demonstrated that the deficits observed were not attributable to nonassociative changes that might have artifactually skewed the data. Even severely impaired 36-month-old rabbits were able to reach a criterion of 80% CRs when switched from a trace to a delay conditioning task that is not hippocampally dependent. The results are discussed in terms of operationally defining and predicting behavioral effects of aging, hypothetical neural mechanisms, and efficient experimental design.

The interaction between nitrous oxide and isoflurane on suppression of learning: a study using classical conditioning in rabbits.

Nitrous oxide (N2O) is commonly combined with a volatile agent for administration of general anesthesia. We studied the effects of N2O and isoflurane on learning of the rabbit nictitating membrane responses (NMRs). Classical conditioning of the NMR was accomplished by presenting a 400 ms tone conditioned stimulus before the presentation of a 100 ms shock unconditioned stimulus over 6 daily training sessions. The percentages of conditioned responses (CRs) were calculated for animals treated with 0% (n = 10), 33% (n = 11), 67% (n = 11), and 75% (n = 7) N2O and for those treated with 0% (n = 8), 0.2% (n = 7), 0.4%, (n = 13) and 0.8% (n = 9) isoflurane separately. ED-50 for suppression of learning for each drug were calculated. Percentages of CRs were calculated for treatments with combinations of 0.2% isoflurane with either 32 or 48% N2O (n = 14, for each). Isobolographic analysis demonstrated that the combination of the two drugs exerted no greater effect than that seen with either agent administered alone; for well-established CRs (mean of days 5 and 6), the estimated concentrations corresponding to a rate of 70% CRs were 0.31% isoflurane with no N2O, 65.3% N2O with no isoflurane, and 0.2% isoflurane combined with 32.4% N2O. N2O and isoflurane interact additively on suppression of learning.

Trace eyeblink conditioning in rabbits demonstrates heterogeneity of learning ability both between and within age groups

Rabbits 2 to 41 months of age were conditioned in the 500 ms trace eyeblink paradigm to cross-sectionally define the age of onset and the severity of age-associated impairments in acquisition of this relatively difficult hippocampally dependent task. Using a strict behavioral criterion of 80% conditioned responses (CRs), age-associated learning impairments were significant by 24 months of age. Among rabbits that successfully reached this criterion, impairments in acquisition plateaued at 30 months of age. However, the proportion of severely impaired rabbits (that failed to reach the 80% criterion) continued to increase age dependently. Using an easier criterion of 8 out of 10 CRs, behavioral impairments were not detected until 30 months of age, and cases of severe impairment (failure to reach criterion) were rare. Additional controls demonstrated that the deficits observed were not attributable to nonassociative changes that might have artifactually skewed the data. Even severely impaired 36-month-old rabbits were able to reach a criterion of 80% CRs when switched from a trace to a delay conditioning task that is not hippocampally dependent. The results are discussed in terms of operationally defining and predicting behavioral effects of aging, hypothetical neural mechanisms, and efficient experimental design.

Alterations in Condition of Cottontail Rabbits (Sylvilagus floridanus) on Rangelands following Brush Management

Although the use of herbicides and prescribed fire have been shown to increase density of cottontail rabbit (Sylvilagus floridanus) populations, the impact of such brush management practices on their condition has not been explored. We used discriminant analysis to investigate responses of overall physical condition of cottontail rabbits (n = 422 adults) to brush management and succession on replicated disturbed and undisturbed upland hardwood forest-tallgrass prairie over a 6-year period. Five different disturbed habitat types were experimentally created using herbicides (tebuthiuron or triclopyr), fire, or a combination of both. Parameters that were important discriminators of rabbit physical condition among habitat types and post-disturbance successional changes included indices of kidney fat and parasitism, and relative masses of spleen, liver, and dried stomach digesta. Brush management practices using herbicides influenced overall condition of rabbits, but the type of habitat disturbance was not important. Effects on overall body condition of cottontail rabbits from burning disturbed habitats were not apparent until later seral stages when production of herbaceous dicots declined and vegetative composition more closely resembled that of undisturbed areas.

Multiple-unit activity from rabbit cerebellar cortex and interpositus nucleus during classical discrimination/reversal eyelid conditioning

Multiple-unit activity was monitored in the hemispheric lobule VI (HVI) and the interpositus nucleus of the cerebellum during classical discrimination and reversal eyelid conditioning in rabbits. During both phases of conditioning similar patterns of activity were observed in lobule HVI: conditioned response (CR)-related activity was present on conditioned stimulus (CS)+ trials but not on CS — trials. Activity in the interpositus nucleus differed during the two phases of conditioning. After discrimination training, CR-related activity was present on CS + but not CS— trials. After reversal training, however, only a weak activation of the interpositus nucleus was seen on CS + trials with no activation present on CS — trials. These data suggest that lobule HVI and the interpositus nucleus may play different roles in classical eyelid conditioning.

Multiple- and single-unit activity in area 32 (prelimbic region) of the medial prefrontal cortex during Pavlovian heart rate conditioning in rabbits.

Data from both brain lesion and brain stimulation experiments suggest that area 32 of the medial prefrontal cortex (prelimbic area) participates in the acquisition and/or expression of conditioned bradycardia. This report describes experiments designed to determine whether cells in this area of the brain exhibit changes in neural activity during classical conditioning that can be related to these learned heart rate changes. Thus, multiple- and single-unit activity was recorded from area 32 in rabbits during Pavlovian heart rate (HR) conditioning. In the first experiment, neuronal discharge recorded from chronically implanted multiple-unit electrodes in the superficial and deep layers of area 32 increased systematically in response to the presentation of tone conditioned stimuli (CS) paired with paraorbital electric shock as the unconditioned stimulus (US). This tone-evoked increase in multiple-unit activity (MUA) closely paralleled the acquisition of the decelerative HR conditioned response in animals that received paired CS/US presentations, but was of smaller amplitude in animals given unpaired CS/US presentations. Tone-evoked increases in MUA also occurred during tone-alone presentations prior to training, but this activity declined over trials. During extinction, CS-evoked MUA increases also declined over trials. These findings suggest that the CS-evoked increase in neuronal activity in area 32 was associatively produced. A second experiment examined the CS-evoked response of single units (n = 98) in area 32 during differential Pavlovian conditioning, in which one tone (CS+) was consistently followed by the paraorbital shock US and a second tone (CS-) was not. Four types of cells were found: (1) 32% of the cells studied showed no change in response to either CS+ or CS- presentation, (2) 48% showed CS-evoked increases to either CS+ or CS-, (3) 14% showed CS-evoked decreases to CS+ or CS-, and (4) 7% showed a biphasic response, in which an increase was followed by a decrease in activity. These cell types are similar to those previously found in the more dorsal anterior cingulate cortex (area 24) and frontal eyefields (area 8). However, a major difference between the present findings and those of our previous studies of areas 8 and 24 is that in several instances changes in CS-evoked activity in area 32 were significantly greater in response to the CS- than to the CS+, suggesting that some cells in the prelimbic region may code the absence of aversive stimulation, that is, a period of relative "safety."

Nefiracetam (DM-9384): effect on eyeblink classical conditioning in older rabbits.

The nootropic compound nefiracetam was evaluated in 88 older rabbits in a 750-ms delay paradigm of eyeblink classical conditioning (EBCC). Rabbits (mean age = 28.7 months) were assigned in groups of eight to one of six conditions in experiment 1: paired tone conditioned stimulus (CS)-corneal airpuff unconditioned stimulus (US) presentations and 1, 3, or 10 mg nefiracetam/kg or sterile saline vehicle; explicitly unpaired CS and US presentations and 3 mg nefiracetam/kg or sterile saline vehicle. Animals in the paired conditions received 10 daily sessions of 90 paired trials and animals in the unpaired conditions received 10 daily sessions of 180 unpaired CS and US presentations. The six conditions in experiment 2 were 5, 10, and 15 mg nefiracetam/kg and vehicle in 15 sessions of paired presentations; 10 mg nefiracetam/kg and vehicle in 15 sessions of unpaired conditioning. In both experiments 1 and 2, acquisition measured by trials to learning criterion was significantly faster at the 10 mg/kg dose of nefiracetam. In the repeated measures analyses comparing six doses in the paired conditions, all dependent measures [percentage conditioned responses (CRs), CR amplitude, and response latency] indicated significantly better conditioning in rabbits treated with 10 mg nefiracetam/kg, but this dose did not increase motor responding or responding in the unpaired condition. Nefiracetam facilitated acquisition of EBCC in older rabbits. EBCC is performed poorly by older humans and is seriously impaired in Alzheimer's disease. These preclinical data in an animal model with clear parallels in humans suggest that nefiracetam may prove effective as a cognition enhancer in clinical trials.