This paper was supported by CAS Key Laboratory of Mental Health, Institute of Psychology (KLMH2016K01) and Evaluation and Intervention Technology Research for Post-traumatic Stress Patients Population (JCYJ20170413170301569)

This study is approved by the International Review Board (IRB) of the Institute of Psychology, Chinese Academy of Sciences, and all experiments are conducted in accordance with the National Institutes of Health (U.S.A) Guide for Care and Use of Laboratory Animals (2011).
1. Materials and Setup for the Conflict Model
<ol>
	<li>House four rats per cage (50 cm long x 22.5 cm wide x 30 cm high) in colony rooms with controlled temperature (22&#8211;25 &#176;C) and a reversed 12 h/12 h light/dark cycle (lights on at 21:00) for at least 10 days. 
	NOTE: Male and female Sprague-Dawley rats weighing 330&#8211;400 and 230&#8211;250 g at the beginning of the experiments, respectively were used.</li>
	<li>Perform all the tests under dim lighting during the dark phase.</li>
	<li>Use the opaque cartons (37 cm long x 26 cm wide x 18 cm high) with pine wood shaving bedding to transport rats from home cages to testing rooms.</li>
	<li>Handle rats 3 min each per day for 5 days prior to beginning the experiments.</li>
	<li>Prepare 120 mL of morphine for the whole experiment. Dissolve morphine hydrochloride in 0.9% NaCl (physiological saline) at a final concentration of 20 mg/mL. Store at room temperature (RT).</li>
	<li>Prepare 10 mL of the pentobarbital sodium solution for the surgery. Dissolve pentobarbital sodium in 0.9% NaCl at a concentration of 1 g/mL. Store at 4 &#176;C.</li>
	<li>Prepare 10 mL of estradiol benzoate (EB) and 10 mL of progesterone for artificial induction of estrus. Dissolve estradiol benzoate and progesterone in sesame oil at a concentration of 0.125 mg/mL and 5 mg/mL, respectively. 
	NOTE: Incubate the oil suspension of EB or progesterone in a water bath (55&#8211;60 &#176;C) for at least 1 h and then thoroughly shake it. Ensure that EB or progesterone is dissolved completely. Store at RT.</li>
	<li>For the risky reward-seeking behavior testing, use the open-field reward-proximity chambers made of black acrylic glass. At one end of the open-field arena (85 cm long x 35 cm wide x 50 cm high) mount a wire-screen stimulus cage (15 cm long x 25 cm wide x 25 cm high). Install the board with pins on the floor about 20 cm in front of the stimulus-cage. 
	NOTE: Pins are fixed on the 34.5 cm long x 13 cm wide boards (Figure 1). The boards with three types of pins are used in turn during testing (Table 2).</li>
</ol>
2. Estrus induction in Females and Mating Screening in Males
<ol>
	<li>Bilateral ovariectomy in female rats 
	NOTE: The surgical instruments and other items used in surgery are sterile. And surgery is performed using aseptic technique.
	<ol>
		<li>Prepare surgical instruments and materials such as scalpels, surgical blades, hemostatic forceps, tweezers, ophthalmic scissors, gauze, cotton swabs, suture needles and silk sutures in advance, as well as 75% alcohol, iodine, sodium pentobarbital, 0.9% NaCl, penicillin sodium.</li>
		<li>Intraperitoneally inject pentobarbital sodium (55 mg/kg), waiting for complete anesthesia of female rats. Rats respire smoothly and do not react to tail pinch. 
		NOTE: The ovariectomy is performed only when the weight of female rat reaches at least 240 g.</li>
		<li>Place the female rat in prone position, shave off the fur on the back, and disinfect the exposed skin with iodophor. Then use a scalpel to make a middle vertical incision (about 2 cm long) on the back (1 cm below the edge of the ribcage).</li>
		<li>Pull the skin to the left side, and blunt dissect subcutaneous tissue with hemostatic forceps to expose the lumbar muscles. Cut the muscle layer (incision of 1 cm) into the abdominal cavity, till the adipose tissue is reached.</li>
		<li>Pull out the adipose tissue with bent tweezers, and locate the ovary which is a flesh-pink tissue (about 0.5 cm x 0.4 cm x 0.3 cm) surrounded by adipose tissue with the twining fallopian tube on the surface.</li>
		<li>Clamp the fallopian tube with forceps and ligate it, then cut off the ovary together with the surrounding adipose tissue.</li>
		<li>Put the residual fallopian tube and adipose tissue back into the abdominal cavity after ensuring no bleeding, and suture the muscle layer.</li>
		<li>Remove the ovary on the other side by the same procedure and then suture the skin incision.</li>
		<li>Place the rat on a heated blanket till awakening and then put it back into the home cage.</li>
		<li>After at least two weeks of recovery, use the female rats as a tool for mating screening and behavioral testing. 
		NOTE: Pay attention to the state of the female rat after surgery, and supply adequate water and food. Keep the ovariectomized rats single housed for one week and then house four per cage.</li>
	</ol>
	</li>
	<li>Induction of estrus in ovariectomized female rats
	<ol>
		<li>Handle all ovariectomized female rats three times (3 min/day) before using them for mating screening.
		<ol>
			<li>Gently pick up a rat from the transport box with the left hand and hold it in the arms for a few seconds. Then put the rat back into the box and repeat these operations for 3 min.</li>
		</ol>
		</li>
		<li>Subcutaneously inject estradiol benzoate (25 &#956;g/rat) about 48&#8211;52 h before mating screening or the conflict test.</li>
		<li>Subcutaneously inject progesterone (1 mg/rat) about 4&#8211;6 h before mating screening or the conflict test. 
		NOTE: The estrogenic hormones are injected subcutaneously on the back of the neck. Since an estrus cycle lasts for ~4&#8210;5 days, the female rats are used once a week.</li>
	</ol>
	</li>
	<li>Screening male rats for mating performance 
	NOTE: Screening is conducted under dim light during the dark phase of the light/dark cycle in the housing room.
	<ol>
		<li>Place a male rat individually into the carton (60 cm long x 50 cm wide x 40 cm high) with pine wood shaving bedding and leave it to habituate for 5 min.</li>
		<li>Introduce an estrous female rat into the carton and monitor male copulatory behaviors (by experienced observers).</li>
		<li>Put the rats back to home cages after male rats complete their first ejaculation within 30 min or do not display intromission within 15 min or ejaculation within 30 min.</li>
		<li>Assign the male rats that pass the screening (successful ejaculation for three consecutive days) randomly into different groups (such as the saline- and morphine-treated groups).</li>
	</ol>
	</li>
</ol>
3. Pretreatment in Male Rats Prior to the Conflict Test
<ol>
	<li>Binge-like morphine treatment 
	NOTE: Male rats are intraperitoneally injected with saline or morphine delivered in a binge-like regimen14 (Table 1).
	<ol>
		<li>Weigh the male rats and calculate the injection volume for each rat based on the body weight (see Table 1).</li>
		<li>Prepare syringes with morphine or saline solutions.</li>
		<li>Inject one rat at a time intraperitoneally and immediately place it gently into the home cage (4 rats/cage).</li>
		<li>After at least 6 h, give the male rats the second injection in the same way.</li>
	</ol>
	</li>
	<li>Acute stress 
	NOTE: Foot shocks are delivered prior to each conflict test in four identical chambers assembled with four shock generators and controlled by professional software installed on a computer.
	<ol>
		<li>On the day of the conflict test, take the male rats to another room different from the conflict testing room.</li>
		<li>Put the male rats into the chambers (30.5 cm long x 25.4 cm wide x 30.5 cm high) to habituate for 1 min.</li>
		<li>Set up the software program in advance. The program includes the intermittent foot shocks delivered within 10 min (0.5 mA x 0.5 s x 10 min; mean inter-shock interval 40 s, range 10&#8211;70 s).</li>
		<li>Enter animal IDs and choose whether to turn on the shock generators according to the grouping (the shock group and the control group). Then press the start button.</li>
		<li>When the stress procedure is finished, bring the rats immediately to the conflict test room in transport boxes; one rat per box.</li>
	</ol>
	</li>
</ol>
4. The Conflict Test
NOTE: The test is conducted under dim light during the dark phase of the light/dark cycle in the conflict test room.
<ol>
	<li>On the day before testing, bring all rats to the conflict test room and allow them to habituate to the open-field reward-proximity chamber (without any obstacle, Figure 1) for 15 min.</li>
	<li>On the testing day, place the male rat in the chamber allowing free exploration for 10 min (under the same condition as the day before testing).</li>
	<li>Place an estrous female rat in the stimulus-cage as an incentive and allow the male subject to freely approach and investigate the incentive rat for 5 min. 
	NOTE: The male rat is randomly exposed to an estrous female, and this female is not a familiar rat.</li>
	<li>After 5 min-free approach, move the male subject from the stimulus-cage to the other end of the arena, place an obstacle (a 14 cm-wide board thick with pins), and then start the first trial of the test. 
	NOTE: The difficulty level of obstacles is varied among trials based on the types of pins and the height of the board. The grading system is shown in Table 2.</li>
	<li>Move the male rat away from the stimulus-cage about 15&#8211;20 s after each time it surmounts the obstacle.</li>
	<li>End a trial if the male subject climbs or jumps over the obstacle 3 times within 4 min and immediately start the next trial with increasing difficulty of the obstacle.</li>
	<li>If a male subject surmounts the obstacle less than three times within 4 min, end the test and record the times it surmounts the obstacle.</li>
	<li>Bring the male rat back to the home cage and scrub the open-field chamber with 0.05% glacial acetic acid.</li>
	<li>Score each surmounting (or approaching) as per Table 2. Use the sum of the scores for all of the surmountings as a total score for a male subject in this conflict test.</li>
</ol>
5. Statistical Analysis
<ol>
	<li>Present the data as mean &#177; SEM or single data points. In case that homogeneity of variance or normal distribution of the datasets is challenged, log-transform the data sets.</li>
	<li>Analyze the effects of morphine pretreatment on reward-seeking behavior displayed on day 7 and day 17 of withdrawal (Wd7 and Wd17) using t-tests with &#8220;pretreatment&#8221; as a between-subjects factor (morphine versus saline, Figure 2).</li>
	<li>Analyze the effects of morphine pretreatment on reward-seeking behavior after repeated testing using repeated-measure&#8217;s analysis of variance (ANOVA) with &#8220;withdrawal time&#8221; (Wd7 versus Wd14) as the within-subjects factor and &#8220;pretreatment&#8221; (morphine versus saline) as a between-subjects factor.</li>
	<li>In addition, use Pearson&#8217;s correlation to analyze the correlation between the scores that male subjects acquired on days 7 and 14 of abstinence (Wd7 and Wd14, Figure 3). 
	NOTE: After the logarithmic conversion of the original data, statistical analyses are performed.</li>
	<li>Analyze the effect of acute stress on reward-seeking behavior in drug-na&#239;ve rats using t-test with stress as a between-subjects factor (shock versus control, Figure 4)</li>
</ol>

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The authors have nothing to disclose.

The inhibitory control deficits caused by drug abuse18 play a key role in promoting compulsive drug seeking/taking behaviors and relapse19,20. The conflict model presented here provides a new approach to explore the changes in inhibitory control of the individuals exposed to addictive drugs.
There are several critical steps in the protocol. First of all, the subjects (male rats) must acquire sexual experience be...

Drug addiction is a chronic brain disease which is characterized by impulsive and compulsive drug seeking and taking1. These key features of addiction have both been hypothesized to result from the impaired ability of inhibitory control2,3, i.e., failing in inhibiting the immediate pursuit of rewarding stimuli and thus developing maladaptive patterns of behavior4.
The go/no-go task and stop-signal task are the prototypical tasks used to measure the ability of response inhibition2,5. These two experimental paradigms assess one&#39;s ability to suppress actions that are inappropriate, by contrasting infrequent inhibitory responses against an implicit go baseline6,7. The response inhibition displayed in these tasks has been shown to be impaired in cocaine users8,9, opiate addicts10, and nicotine users11. Another two tasks&#8212;reversal learning and multiple choice serial reaction time tasks&#8212;also provide measurements of response inhibition/inhibitory control12,13. However, most of these paradigms performed in rodents not only require long-term training so that subjects can distinguish the response requirements represented by different signals, but also the individual differences in learning speed and learning effects may interfere with the results of subsequent inhibitory test11.
In this paper, we present a novel conflict task which can be used to measure the impairment of inhibitory control after exposure to addictive drugs. In this task, a natural rewarding stimulus (sexual stimulus) that represents a high-value reward14, and the aversive stimuli (pins) that male rats have to conquer, are concurrently presented. The male rats have to climb or jump over the obstacle full of pins to approach and investigate the sexual partner. If the animal persists in its approaching behavior regardless of the aversive stimuli, it is considered as a maladaptive or risky reward-seeking behavior. One of the rationales for establishing this task is that it is conceptually simple and does not place heavy demands on executive processes as other tasks do. Compared to other tasks which measure response inhibition, this conflict task is based on natural behavior, and rats with normal sexual function and sexual experience can be tested directly without a learning process. Another rationale is that a conflict presented in this task between approaching the reward and avoiding the aversive stimuli (or the risk of being pricked) may have a better validity, as it mimics what occurs in addicts who often place themselves in the similar conflict but persistently pursue drug reward regardless of the risk of negative consequences in real life15.
Therefore, application of this conflict model is a quick and sensitive way to discover the deficit in inhibitory control after exposure to addictive drugs, or other factors that may influence ability of inhibitory control, such as stress. It also provides a novel behavioral strategy for investigation of neural mechanisms underlying deficits in inhibitory control. Furthermore, alternative modifications can be added onto this task. For example, altering the cost/benefit ratio by replacing the sexual stimulus with the social stimulus can reveal more behavioral significances.

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a conflict model of reward-seeking behavior in male rats

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual stimulus) that represents a high-value reward, and the aversive stimuli (pins), are concurrently presented. The male rats have to climb or jump over the obstacle full of pins to approach and investigate the sexual partner. If the animal persists in their approaching behavior regardless of the aversive stimuli, it is considered as a maladaptive or risky reward-seeking behavior. The conflict task permits the evaluation of deficit in inhibitory control resulting from exposure to abused drug, such as morphine, or a stressful event. 
The main advantage of this model is that it provides a simple and quick way to discover the deficit in inhibitory control after exposure to opiate drugs or other stressful events. In addition to opiates, this behavioral model would also be useful for quickly discovering the inhibitory control deficits induced by other addictive drugs. However, the limitation is that the male rats' performance may be subject to exercising effects with repeated testing under this conflict task. In the future, one can hope that the individuals with the compulsive phenotype of reward-seeking behavior after exposure to opiates will be identified based on modifying this conflict model.

To explore whether this conflict model can reveal maladaptive/risky reward-seeking behavior induced by opiates, the reward-seeking behaviors displayed by the saline- and morphine-pretreated groups were compared by t-tests after short-term (Wd7) and long-term (Wd17) withdrawal from morphine respectively (Figure 2). The results show that on both day 7 and day 17 of withdrawal, the morphine-pretreated rats showed significantly more approaching behaviors than the...

Watch this Scientific Journal Video about A Conflict Model of Reward-seeking Behavior in Male Rats at JoVE.com

A Conflict Model of Reward-seeking Behavior in Male Rats

This conflict model is used to measure the impairment of inhibitory control after exposure to addictive drugs, or other factors that may influence inhibitory control. A sexual stimulus and an aversive obstacle are concurrently presented, thus male rats have to conquer the obstacle to approach the sexual reward.

The present protocol describes a novel conflict task as a model of inhibitory control in rats. In this model, a natural rewarding stimulus (sexual ...

a-conflict-model-of-reward-seeking-behavior-in-male-rats

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JoVE Journal

Behavior

7.3K Views.  Chinese Academy of Sciences. This conflict model is used to measure the impairment of inhibitory control after exposure to addictive drugs, or other factors that may influence inhibitory control. A sexual stimulus and an aversive obstacle are concurrently presented, thus male rats have to conquer the obstacle to approach the sexual reward. 

Video: A Conflict Model of Reward-seeking Behavior in Male Rats

A Procedure to Study the Effect of Prolonged Food Restriction on Heroin Seeking in Abstinent Rats

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during abstinence. Moreover, various environmental challenges can exacerbate this effect, as well as increase ongoing drug intake.
The procedure we describe here highlights the impact of a common environmental challenge, food restriction, on drug craving that is expressed as an augmentation of drug seeking in abstinent rats.
Rats are implanted with chronic intravenous i.v. catheters, and then trained to press a lever for i.v. heroin over a period of 10-12 days. Following the heroin self-administration phase the rats are removed from the operant conditioning chambers and housed in the animal care facility for a period of at least 14 days. While one group is maintained under unrestricted access to food (sated group), a second group (FDR group) is exposed to a mild food restriction regimen that results in their body weights maintained at 90% of their nonrestricted body weight. On day 14 of food restriction the rats are transferred back to the drug-training environment, and a drug-seeking test is run under extinction conditions (i.e.&#160;lever presses do not result in heroin delivery).
The procedure presented here results in a highly robust augmentation of heroin seeking on test day in the food restricted rats. In addition, compared to the acute food deprivation manipulations we have used before, the current procedure is a more clinically relevant model for the impact of caloric restriction on drug seeking. Moreover, it might be closer to the human condition as the rats are not required to go through an extinction-training phase before the drug-seeking test, which is an integral component of the popular reinstatement procedure.

A procedure that allows a demonstration of robust augmentation of drug seeking in food-restricted rats is described. Following heroin self-administration training, rats go through an abstinence period, in a drug-free environment, during which they are mildly food restricted. Drug seeking is then tested in the drug-associated environment.

In human drug addicts, exposure to drug-associated cues or environments that were previously associated with drug taking can trigger relapse during ...

Fat Preference: A Novel Model of Eating Behavior in Rats

Obesity is a growing problem in the United States of America, with more than a third of the population classified as obese. One factor contributing to this multifactorial disorder is the consumption of a high fat diet, a behavior that has been shown to increase both caloric intake and body fat content. However, the elements regulating preference for high fat food over other foods remain understudied.
To overcome this deficit, a model to quickly and easily test changes in the preference for dietary fat was developed. The Fat Preference model presents rats with a series of choices between foods with differing fat content. Like humans, rats have a natural bias toward consuming high fat food, making the rat model ideal for translational studies. Changes in preference can be ascribed to the effect of either genetic differences or pharmacological interventions. This model allows for the exploration of determinates of fat preference and screening pharmacotherapeutic agents that influence acquisition of obesity.

Dietary fat content influences both energy intake and body fat composition in mammals. By examining rats&#8217; preference for high fat food in a series of choice experiments, it is possible to test genetic differences and pharmacological interventions on their preference for high fat food.

Obesity is a growing problem in the United States of America, with more than a third of the population classified as obese. One factor contributing to ...

A Procedure to Observe Context-induced Renewal of Pavlovian-conditioned Alcohol-seeking Behavior in Rats

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate drug-seeking behavior and prompt relapse in abstinent drug users. We have developed a procedure to study the behavioral and neural processes that mediate the impact of context on alcohol-seeking behavior in rats. Following acclimation to the taste and pharmacological effects of 15% ethanol in the home cage, male Long-Evans rats receive Pavlovian discrimination training (PDT) in conditioning chambers. In each daily (Mon-Fri) PDT session, 16 trials each of two different 10 sec auditory conditioned stimuli occur. During one stimulus, the CS+, 0.2 ml of 15% ethanol is delivered into a fluid port for oral consumption. The second stimulus, the CS-, is not paired with ethanol. Across sessions, entries into the fluid port during the CS+ increase, whereas entries during the CS- stabilize at a lower level, indicating that a predictive association between the CS+ and ethanol is acquired. During PDT each chamber is equipped with a specific configuration of visual, olfactory and tactile contextual stimuli. Following PDT, extinction training is conducted in the same chamber that is now equipped with a different configuration of contextual stimuli. The CS+ and CS- are presented as before, but ethanol is withheld, which causes a gradual decline in port entries during the CS+. At test, rats are placed back into the PDT context and presented with the CS+ and CS- as before, but without ethanol. This manipulation triggers a robust and selective increase in the number of port entries made during the alcohol predictive CS+, with no change in responding during the CS-. This effect, referred to as context-induced renewal, illustrates the powerful capacity of contexts associated with alcohol consumption to stimulate alcohol-seeking behavior in response to Pavlovian alcohol cues.

A procedure to study the capacity of an alcohol associated environmental context to trigger the renewal of alcohol-seeking behavior in rats is described.

Environmental contexts in which drugs of abuse are consumed can trigger craving, a subjective Pavlovian-conditioned response that can facilitate ...

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure multiple dimensions of unconditioned behavior in small laboratory mammals (e.g., mice, rats, tree shrews and small primates). This paradigm is a valuable alternative for the use of a behavioral test battery, since a broad behavioral spectrum of an animal&#8217;s behavioral profile can be investigated in one single test.
The apparatus consists of a box, representing the &#8216;protected&#8217; area, separated from a group compartment. A board, on which small cylinders are staggered in three lines, is placed in the center of the box, representing the &#8216;unprotected&#8217; area of the set-up. The cognitive abilities of the animals can be measured by baiting some cylinders on the board and measuring the working and reference memory. Other unconditioned behavior, such as activity-related-, anxiety-related- and social behavior, can be observed using this paradigm. Behavioral flexibility and the ability to habituate to a novel environment can additionally be observed by subjecting the animals to multiple trials in the mHB, revealing insight into the animals&#8217; adaptive capacities.
Due to testing order effects in a behavioral test battery, na&#239;ve animals should be used for each individual experiment. By testing multiple behavioral dimensions in a single paradigm and thereby circumventing this issue, the number of experimental animals used is reduced. Furthermore, by avoiding social isolation during testing and without the need to food deprive the animals, the mHB represents a behavioral test system, inducing if any, very low amount of stress.

This protocol describes the modified hole board, which is a behavioral test set-up that comprises the characteristics of an open field and a traditional hole board. This set-up enables the differential analysis of unconditioned behavior of small laboratory mammals as well as the analysis of cognitive abilities.

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

Acquisition of a High-precision Skilled Forelimb Reaching Task in Rats

Movements are the main measurable output of central nervous system function. Developing behavioral paradigms that allow detailed analysis of motor learning and execution is of critical importance in order to understand the principles and processes that underlie motor function. Here we present a paradigm to study movement acquisition within a daily session of training (within-session) representing the fast learning component and primary acquisition as well as skilled motor learning over several training sessions (between-session) representing the slow learning component and consolidation of the learned task. This behavioral paradigm increases the degree of difficulty and complexity of the motor skill task due to two features: First, the animal realigns its body prior to each pellet retrieval forcing renewed orientation and preventing movement execution from the same angle. Second, pellets are grasped from a vertical post that matches the diameter of the pellet and is placed in front of the cage. This requires a precise grasp for successful pellet retrieval and thus prevents simple pulling of the pellet towards the animal. In combination with novel genetics, imaging and electrophysiological technologies, this behavioral method will aid to understand the morphological, anatomical and molecular underpinnings of motor learning and memory.

A paradigm is presented to analyze the acquisition of a high-precision skilled forelimb reaching task in rats.

Movements are the main measurable output of central nervous system function. Developing behavioral paradigms that allow detailed analysis of motor ...

The Rodent Psychomotor Vigilance Test (rPVT): A Method for Assessing Neurobehavioral Performance in Rats and Mice

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article describes a rodent version of the PVT&#8212;termed the &#34;rPVT&#34;&#8212;that measures similar aspects of attention (i.e., performance accuracy, motor speed, premature responding, and lapses in attention). Data are presented that demonstrate both the short- and long-term usefulness of the rPVT when employed with laboratory rats. Rats easily learn the rPVT, and learning to perform the basic procedure takes less than two weeks of training. Once acquired, rat performances in the rPVT show a high degree of similarity to these same performance measures in the human PVT, including similarities in, lapses in attention, reaction times, vigilance decrements across session time (i.e., the human &#34;time-on-task&#34; effects), and the response-stimulus interval (RSI) effect described for humans. Thus the rPVT can be an extremely valuable tool for assessing the effects of a wide range of variables on sustained attention quite similar to human PVT performances, and thus can be useful for developing novel treatments for neurobehavioral dysfunctions.

The Rodent Psychomotor Vigilance Test rPVT: A Method for Assessing Neurobehavioral Performance in Rats and Mice

A rat version of the human Psychomotor Vigilance Test (PVT) is described that measures aspects of attention similar to those measured with the human PVT, including aspects of human vigilance such as performance accuracy, motor speed, premature responding, and lapses in attention.

The human Psychomotor Vigilance Test (PVT) is a widely used procedure for measuring changes in fatigue and sustained attention. The present article ...

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, few studies have investigated the long-term effects of injury in rodent models of TBI. Shown here are behavioral measures that are well-established in TBI research for times early after injury, such as two weeks, until two months. Some of these methods have previously been used at later times after injury, up to one year, but by very few laboratories. The methods demonstrated here are a short neurological assessment to test reflexes, a Beam-Balance to test balance, a Beam-Walk to test balance and motor coordination, and a working memory version of the Morris water maze that can be sensitive to deficits in reference memory. Male rats were handled and pre-trained to neurological, balance, and motor coordination tests prior to receiving parasagittal fluid percussion injury (FPI) or sham injury. Rats can be tested on the short neurological assessment (neuroscore), the beam-balance, and the Beam-Walk multiple times, while testing on the water maze can only be done once. This difference is because rats can remember the task, thus confounding the results if repeated testing is attempted in the same animal. When testing from one to three days after injury, significant differences are detected in all three non-cognitive tasks. However, differences in the Beam-Walk task were not detectable at later time points (after 3 months). Deficits were detected at 3 months in the Beam-Balance and at 6 months in the neuroscore. Deficits in working memory were detected out to 12 months after injury, and a deficit in a reference memory first appeared at 12 months. Thus, standard behavioral tests can be useful measures of persistent behavioral deficits after FPI.

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after injury.

With the increasing incidence of traumatic brain injury (TBI) in both civilian and military populations, TBI is now considered a chronic disease; however, ...

A New Method for Inducing a Depression-Like Behavior in Rats

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is no existing format for studying the mechanism of action, prevention, containment, and treatment of contagious depression. The purpose of this study, therefore, was to establish the first animal model of contagious depression.
Healthy rats can contract depressive behaviors if exposed to depressed rats. Depression is induced in rats by subjecting them to several manipulations of chronic unpredictable stress (CUS) over 5 weeks, as described in the protocol. A successful sucrose preference test confirmed the development of depression in the rats. The CUS-exposed rats were then caged with na&#239;ve rats from the contagion group (1 na&#239;ve rat/2 depressed rats in a cage) for an additional 5 weeks. 30 social groups were created from the combination of CUS-exposed rats and na&#239;ve rats.
This proposed depression-contagion protocol in animals consists mainly of cohabiting CUS-exposed and healthy rats for 5 weeks. To ensure that this method works, a series of tests are carried out - first, the sucrose preference test upon inducing depression to rats, then, the sucrose preference test, alongside the open field and forced-swim tests at the end of the cohabitation period. Throughout the experiment, rats are given tags and are always returned to their cages after each test.
A few limitations to this method are the weak differences recorded between the experimental and control groups in the sucrose preference test and the irreversible traumatic outcome of the forced swim test. These may be worth considering for suitability before any future application of the protocol. Nonetheless, following the experiment, na&#239;ve rats developed contagion depression after 5 weeks of sharing the same cage with the CUS-exposed rats.

This protocol describes a new model by which healthy rats could contract depression over a given time periodthrough contagion by exposure to chronic unpredictable stressed (CUS) rats.

Contagious depression is a phenomenon that is yet to be fully recognized and this stems from insufficient material on the subject. At the moment, there is ...

A Conditioned Place Preference Protocol for Measuring Incubation of Craving in Rats

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed incubation of drug craving. Here, we describe a morphine conditioned place preference (CPP) protocol for measuring the incubation of craving in rats. In this protocol, a CPP paradigm mainly employing somatosensory cues is used to establish a long-term reward memory of morphine. A three-chamber CPP box that differs in the texture of the chamber floor is constructed. First, the animals are tested for their baseline preference to the two side chambers for three consecutive days. Then, they are injected intraperitoneally with morphine/saline and put into their non-preferred/preferred chamber for 45 min. After 6 days of conditioning, their preference to the side chambers is tested for 15 min at different time points after the last conditioning session. With this paradigm, the reward memory of morphine could last for at least 18 days. To test whether the above-mentioned protocol can model increased craving, the number of entrances into the two side chambers are counted during the abstinence period. The results show that the entrances increased, suggesting that the CPP paradigm could mimic the incubation of craving. Future studies can employ this model to study neural mechanisms underlying long-term memory and incubation of craving.

Here, a morphine conditioned place preference (CPP) protocol is described to measure incubation of craving in rats.

A major cause of repeated relapses is a craving for the drug. Drug craving increases progressively during the abstinence period, a phenomenon termed ...

A Laboratory Method to Measure Contagious Yawning in Rats

Communication is an essential aspect of animal social life. Animals may influence one another and come together in schools, flocks, and herds. Communication is also the way sexes interact during courtship and how rivals settle disputes without fighting. However, there are some behavioral patterns for which it is difficult to test the existence of a communicatory function, because several types of sensory modalities are likely involved. For example, contagious yawning is a communicatory act in mammals that potentially occurs through sight, hearing, smell, or a combination of these senses depending on whether the animals are familiar to one another. Therefore, to test hypotheses about the possible communicatory role of such behaviors, a suitable method is necessary to identify the participating sensory modalities.
The method proposed here aims to obtain yawn contagion curves for familiar and unfamiliar rats and evaluate the relative participation of visual and olfactory sensory modalities. The method uses inexpensive materials, and with some minor changes, it can also be used with other rodent species such as mice. Overall, the method involves the substitution of clear dividers (with or without holes) with opaque dividers (with or without holes) that either allow or prevent communication between rats placed in adjacent cages with holes in adjoining sides. Accordingly, four conditions can be tested: olfactory communication, visual communication, both visual and olfactory communication, and neither visual nor olfactory communication. As social interaction occurs between the rats, these test conditions simulate what may occur in a natural environment. In this respect, the method proposed here is more effective than traditional methods that rely on video presentations whose biological validity can raise concerns. Nonetheless, it does not discriminate between the potential role of hearing and roles of smell and vision in yawn contagion.

The method described here aims to obtain yawn contagion curves in pairs of familiar or unfamiliar male rats. Cages with holes separated by either clear or opaque partitions with (or without) holes are used to detect whether visual, olfactory, or both types of sensory cues can stimulate yawn contagion.