We would like to dedicate this work to the memory of late Dr. William T. Greenough, a great mentor, a colleague and a friend. This work was supported by NIH/NIAAA grant number AA009838 and NIH/NIGMS COBRE: The Delaware Center for Neuroscience research grant 1P20GM103653 to AYK. We are grateful to the former and current members of Klintsova lab.

Ethics Statement: The following protocol was approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Delaware.
1. Developmental Exposure (or Model of Binge-like Ethanol Exposure)
<ol>
	<li>On PD3, determine the sex of each animal and cross-foster any animals if necessary to keep litter size (8 animals) and sex distribution (4 males : 4 females) consistent within each litter. 
	NOTE: It is important to keep litter size and sex distribution as consistent as possible to avoid experimental confounds. Although this protocol uses 8 pups (4 males and 4 females) per litter, alternative litter sizes or sex distributions may be tailored to the needs of the experimental design.</li>
	<li>Subcutaneously inject a small amount of black India ink into the paws to identify animals within each litter.</li>
	<li>Pseudo-randomly assign litters as experimental (containing 50% alcohol-exposed (AE) and 50% sham-intubated control (SI) pups) or suckle control (SC) (animals that do not undergo any intubation, tail clipping, or separation protocols from PD 4 - 9 except for daily weighing and ear-punching).
	<ol>
		<li>To retain consistent group size, assign twice as many experimental litters as SC litters.</li>
	</ol>
	</li>
	<li>Weigh each animal then return it to its home cage. Animal weighing should occur daily during the intubation period (PD 4 - 9).
	<ol>
		<li>Remove the whole litter from the dam.</li>
		<li>Place pups on heated pad.</li>
		<li>Record the weight of each individual pup.</li>
	</ol>
	</li>
	<li>On PD4, after weighing each animal calculate the necessary alcohol amount for a total of 5.25 g/kg/day per each animal (based on pup weight from step 1.4)8.
	<ol>
		<li>Administer alcohol as 11.9% ethanol-in-milk substitute (vol/vol).</li>
	</ol>
	</li>
	<li>Starting at 9 AM, remove one litter&#39;s pups from the mother at a time.</li>
	<li>Administer the ethanol-in-milk to each AE pup34.</li>
	<li>Sham-intubate each SI pup8.</li>
	<li>Repeat steps 1.5. through 1.8. for each experimental litter.</li>
	<li>Two hours following the first dose, repeat the dosing procedure (steps 1.5 through 1.8) for a second alcohol dose.</li>
	<li>One and a half hours after the second alcohol dose (the point at which peak daily blood alcohol content is achieved), collect and centrifuge blood from the AE and SI pups via tail clipping for future blood alcohol content analysis35.
	<ol>
		<li>Collect 60 &#181;L of blood.</li>
		<li>Place blood in a 1 mL microcentrifuge tube. Centrifuge blood at 1.5 x g for 25 min.</li>
		<li>Carefully collect the supernatant serum from the centrifuge tube and save for future blood alcohol content analysis.</li>
	</ol>
	</li>
	<li>Repeat the dosing procedure (steps 1.5 through 1.8) using milk instead of ethanol-in-milk to prevent nutritional deficits from nursing inability in AE pups.
	<ol>
		<li>Perform a total of 2 supplemental milk doses 2 h apart on PD 4.</li>
	</ol>
	</li>
	<li>Repeat steps 1.4 through 1.12 (except for step 1.11) on PD 5 &#8211; 9.</li>
	<li>Following the final supplemental milk dose on PD9, ear punch all pups for identification in the EC cage.
	<ol>
		<li>Coordinate punched ear with some measure of litter number or identifier (e.g., odd numbered litters within a cohort would get their left ear punched while animals from even numbered litters would get their right ears punched). This will make it easier to identify animals in the EC cage should multiple animals from different litters have the same pawmark pattern.</li>
	</ol>
	</li>
</ol>
2. Weaning
<ol>
	<li>On PD 23, house all animals in cages of 2 - 3.
	<ol>
		<li>Ensure that all animals housed in the same cage are the same sex.</li>
		<li>Include one SC, one SI, and one AE animal per cage when possible.</li>
		<li>Minimize the number of cage mates that are from the same litter.</li>
		<li>Make sure all animals are capable of accessing food and water.</li>
	</ol>
	</li>
</ol>
3. Wheel Running
<ol>
	<li>On PD30, allocate half of the cages with animals to WR. House these animals in cages with a free access to attached stainless steel running wheel.
	<ol>
		<li>Ensure that wheels have a counter to assess the total number of revolutions.</li>
	</ol>
	</li>
	<li>Weigh all animals on PD 30 and PD 36.</li>
	<li>Check the number of revolutions of each wheel at 9 AM every day.</li>
	<li>Leave animals in their respective housing condition for 12 days.</li>
</ol>
4. Environmental Complexity
<ol>
	<li>Prepare the EC cage before 9 AM on the day that corresponds to PD 42 for experimental animals.
	<ol>
		<li>Get a 30 &#34; x 18 &#34; x 36 &#34; galvanized steel cage. 
		NOTE: The cage should have multiple levels, be capable of supporting the weight of multiple rats, be filled with standard bedding, and have multiple locations to attach water bottles and food dispensers.</li>
		<li>Place novel, colorful objects of variable sizes and shapes in the cage.
		<ol>
			<li>Place 6 large toys in the EC cage. Ensure that each toy is big enough for 3 or more rats to interact with concurrently.</li>
			<li>Place 6 medium toys in the EC cage. Ensure that each toy is big enough for 3 - 4 rats to interact with concurrently.</li>
			<li>Place a lot (at least 20) of smaller toys in the EC cage.</li>
			<li>Use toys of varying colors, shapes, size, etc. Novelty is critical to this intervention (see discussion).</li>
		</ol>
		</li>
		<li>Place two dishes of food at opposite ends of the cage.</li>
		<li>Place two bottles of water at opposite ends of the cage.</li>
	</ol>
	</li>
	<li>At 9 AM on PD 42, weigh all animals and relocate the WR animals to the EC cage. Each EC cage should contain 9 - 12 animals.
	<ol>
		<li>Make sure that no animals have both the same pawmark and ear-punch patterns.</li>
	</ol>
	</li>
	<li>Check all food and water daily.</li>
	<li>Every two days, remove the toys from the EC cage and replace them (according to step 4.1.2.).</li>
	<li>Every three days, clean the EC cage.
	<ol>
		<li>Remove the animals from the EC cage and put them in temporary holding cages of 2 - 3 animals.</li>
		<li>Remove all of the bedding from the bottom of the cage.</li>
		<li>Return the same toys to the cage unless this day coincides with the toy replacement schedule (according to step 4.4.).</li>
		<li>Replace all of the food and water.</li>
		<li>Replace the rats into the EC cage.</li>
	</ol>
	</li>
</ol>
5. Collect Tissue
NOTE: Tissue collection (e.g., perfusion with paraformaldehyde) and storage (e.g., freezing, paraffin embedding) can be performed with a variety of methods. The following will explain the process of perfusion with 4% paraformaldehyde in 0.1 M phosphate buffered saline (4% paraformaldehyde in PBS) solution8.
Caution: Paraformaldehyde is carcinogenic and may also cause skin irritation, allergic skin reaction, or eye damage. Use appropriate eye/skin protection.
<ol>
	<li>Expose one rat at a time to isoflurane to lightly anesthetize the animal.</li>
	<li>Intraperitoneally inject the rat with 2 mL/kg of Ketamine/xylazine mixture (1.5 mL xylazine mixed with 10 mL of ketamine). 
	NOTE: Ketamine and xylazine are both at stock concentrations of 100 mg/mL before combining for injection mixture.</li>
	<li>Once rat is no longer responsive, perfuse the animal with 0.1 M phosphate buffered saline (PBS; pH = 7.2) followed by 4% paraformaldehyde in PBS (pH = 7.2).</li>
	<li>Remove brain and store in 4% paraformaldehyde in PBS at 4 &#176;C for 48 h.</li>
	<li>After 2 days, transfer to solution of 30% sucrose added to 4% paraformaldehyde in PBS at 4 &#176;C.</li>
</ol>

The authors have nothing to disclose.

In the above protocol, we demonstrated an expedient intervention to rescue neuroanatomical deficits following neonatal alcohol exposure. This intervention can be used as a therapeutic in other animal models due to the robustness of each of the components of the intervention. Voluntary cardiovascular activity in the form of WR has been shown to benefit several behavioral outcomes38,39 and induce functional plastic alterations in brain regions such as the hippocamp...

Rearing in different environments has long been known to cause changes in various measures of neurological wellness. Many studies look at the beneficial effects of rearing in a complex environment (EC) starting with groundbreaking research by Diamond and Rosenzweig (e.g., 1,2) and Greenough (e.g.,3,4). EC has been demonstrated to have an undeniable positive effects on synaptic and cellular changes in the brain5,6,7. EC can affect a multiplicity of brain regions including the hippocampus8,9 and visual cortex10,11, ventral striatum12,13, as well as brain-wide neuroimmune function (reviewed in14). Particular interest has developed from the studies on hippocampus when it was demonstrated that EC can increase the survival rate of adult-born granule cells of the dentate gyrus through dendritic plasticity 9,13. This last point has gathered much interest due to the growing body of literature indicating that cardiovascular exercise promotes adult neurogenesis in both the healthy and damaged brain15,16,17,18. Wheel running (WR) is an easy to implement form of voluntary cardiovascular activity that has been shown to be beneficial in rodent models of neurological disorders or aging 17,19,20. WR affects the expression of growth factors in both the central and peripheral nervous system 21,22,23.
Combining (subsequently) WR and EC into a &#34;superintervention&#34; (WR-EC) (i.e., 12 days of WR followed by 30 days in EC) provides a robust increase in hippocampal adult neurogenesis and increased survival of the newly proliferated cells8, the effect that in the animal model of FASD is not achieved by individual components (see below). Since both components of WR-EC affect a diverse array of structures within the brain13 (WR reviewed in22, EC reviewed in24), implementation of this intervention can easily be applied to rodent models of both developmental and later life onset models of neurological impairment (e.g., neonatal alcohol exposure, aging, early life stress).
Integration of WR-EC in the adolescent and early adult periods (i.e., postnatal days 30 - 72) can ameliorate some of the negative effects of a rat model of fetal alcohol spectrum disorders (FASDs)8. A collection of studies have demonstrated that rodents exposed to alcohol from postnatal day (PD) 4 through 9 display significant deficits in neuroanatomical measures such as dendritic complexity25, cerebellar development26,27 and neuroimmune responsiveness28 as well as manifestations of impaired learning and memory29,30,31. Even a reduced amount of alcohol exposure within this time window (i.e., PD 7 through 9) can lead to deficits in learning and memory in adolescent and adult rats32 while some structures no longer see significant neuroanatomical impairment27. Many of these deficits - in addition to behavioral impairments in hippocampus-dependent tasks - have been mitigated following exposure to this WR-EC paradigm8,33 or WR alone25,31. Although WR alone has been a widely used intervention, the combination of WR-EC has not yet been utilized in the literature despite its ability to sustain the relatively shorter-term benefits of WR8. This article will discuss the implementation of the WR-EC intervention during adolescence. Although this paradigm is used in the context of early postnatal alcohol exposure, it can be introduced to various rodent models to assess brain potential for neuroplasticity in the models of brain disorders.

<table><tbody><tr><td>Female Time-pregnant Long Evans Rats</td><td>Envigo (Formerly: Harlan, Inc.)</td><td></td><td>Average litter size is 8 - 10 pups</td></tr><tr><td>Black India Ink</td><td>Higgins (Chartpak, Inc.)</td><td>44201</td><td></td></tr><tr><td>Syringes and Injection Needles</td><td>Becton, Dickinson and Company (BD)</td><td>Assorted</td><td>For injection of pawmarking ink, administration of milk-alcohol solution</td></tr><tr><td>Ear Punch</td><td>Kent Scientific Corporation</td><td>INS750076</td><td></td></tr><tr><td>Running Wheels</td><td>Wahmann Labs</td><td></td><td>Wahmann Running Wheel is discontinued. One per cage.</td></tr><tr><td>EC Cage</td><td>Martin&#039;s Cages, Inc.</td><td>R-695</td><td></td></tr><tr><td>Small EC Toys</td><td>Assorted</td><td></td><td></td></tr><tr><td>Medium EC Toys</td><td>Assorted</td><td></td><td>Should be able to fit 1 - 2 rats inside of/on top of object</td></tr><tr><td>Large EC Toys</td><td>Assorted</td><td></td><td>Should be able to fit 3 or more rats inside of/on top of object</td></tr></tbody></table>

wheel running and environmental complexity as a therapeutic intervention in an animal model of fasd

Aerobic exercise (e.g., wheel running (WR) extensively used in animal research) positively impacts many measures of neuroplastic potential in the brain, such as rates of adult neurogenesis, angiogenesis, and expression of neurotrophic factors in rodents. This intervention has also been shown to mitigate behavioral and neuroanatomical aspects of the negative impacts of teratogens (i.e., developmental exposure to alcohol) and age-related neurodegeneration in rodents. Environmental complexity (EC) has been shown to produce numerous neuroplastic benefits in cortical and subcortical structures and can be coupled with wheel running to increase the proliferation and survival of new cells in the adult hippocampus. The combination of these two interventions provides a robust &#34;superintervention&#34; (WR-EC) that can be implemented in a range of rodent models of neurological disorders. We will discuss the implementation of WR/EC and its constituent interventions for use as a more powerful therapeutic intervention in rats using the animal model of prenatal exposure to alcohol in humans. We will also discuss which elements of the procedures are absolutely necessary for the interventions and which ones may be altered depending on the experimenter&#39;s question or facilities.

In order to assess the effect of the super intervention, we must look at the effects of each of its constituent elements &#8212; WR and EC &#8212; on our measures of interest. Figures 1 through 3 (below) appeared in a previous publication utilizing this paradigm8. Figure 4 appeared in a doctoral dissertation36. These data illustrate the impact of WR-EC on hippocampal adult neurogenesis in th...

Watch this Scientific Journal Video about Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD at JoVE.com

Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

Cardiovascular exercise and stimulating experiences in a complex environment have positive benefits on multiple measures of neuroplasticity within the rodent brain. This article will discuss the implementation of these interventions as a &#34;superintervention&#34; which combines wheel running and environmental complexity and will address the limitations of these interventions.

Aerobic exercise (e.g., wheel running (WR) extensively used in animal research) positively impacts many measures of neuroplastic potential in the brain, ...

wheel-running-environmental-complexity-as-therapeutic-intervention-an

Research

JoVE Journal

Behavior

6.7K Views.  University of Delaware. Cardiovascular exercise and stimulating experiences in a complex environment have positive benefits on multiple measures of neuroplasticity within the rodent brain. This article will discuss the implementation of these interventions as a "superintervention" which combines wheel running and environmental complexity and will address the limitations of these interventions.

Video: Wheel Running and Environmental Complexity as a Therapeutic Intervention in an Animal Model of FASD

Using the Activity-based Anorexia Rodent Model to Study the Neurobiological Basis of Anorexia Nervosa

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A challenging illness to treat, due to the lack of understanding of the underlying neurobiology, AN has the highest mortality rate among psychiatric illnesses. To address this need, neuroscientists are using an animal model to study how neural circuits may contribute toward vulnerability to AN and may be affected by AN. Activity-based anorexia (ABA) is a bio-behavioral phenomenon described in rodents that models the key symptoms of anorexia nervosa. When rodents with free access to voluntary exercise on a running wheel experience food restriction, they become hyperactive &#8211; running more than animals with free access to food. Here, we describe the procedures by which ABA is induced in adolescent female C57BL/6 mice. On postnatal day 36 (P36), the animal is housed with access to voluntary exercise on a running wheel. After 4 days of acclimation to the running wheel, on P40, all food is removed from the cage. For the next 3 days, food is returned to the cage (allowing animals free food access) for 2 hr daily. After the fourth day of food restriction, free access to food is returned and the running wheel is removed from the cage to allow the animals to recover. Continuous multi-day analysis of running wheel activity shows that mice become hyperactive within 24 hr following the onset of food restriction. The mice run even during the limited time during which they have access to food. Additionally, the circadian pattern of wheel running becomes disrupted by the experience of food restriction. We have been able to correlate neurobiological changes with various aspects of the animals&#8217; wheel running behavior to implicate particular brain regions and neurochemical changes with resilience and vulnerability to food-restriction induced hyperactivity.

Here we present a protocol to induce activity-based anorexia (ABA) in female adolescent mice. ABA is a condition of hyperactivity evoked by imposing food restriction on rodents with access to a running wheel. This phenomenon is being used as a model to study the underlying neurobiology of anorexia nervosa.

Anorexia nervosa (AN) is a psychiatric illness characterized by excessively restricted caloric intake and abnormally high levels of physical activity. A ...

An Innovative Running Wheel-based Mechanism for Improved Rat Training Performance

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity for reducing the severity of the effects of the stroke&#160;in rats. This system provides more effective animal exercise training than commercially available systems such as treadmills and motorized running wheels (MRWs). In contrast to an MRW that can only achieve speeds below 20 m/min, rats are permitted to run at a stable speed of 30 m/min on a more spacious and high-density rubber running track supported by a 15 cm wide acrylic wheel with a diameter of 55 cm in this work. Using a predefined adaptive acceleration curve, the system not only reduces the operator error but also trains the rats to run persistently until a specified intensity is reached. As a way to evaluate the exercise effectiveness, real-time position of a rat is detected by four pairs of infrared sensors deployed on the running wheel. Once an adaptive acceleration curve is initiated using a microcontroller, the data obtained by the infrared sensors are automatically recorded and analyzed in a computer. For comparison purposes, 3 week training is conducted on rats using a treadmill, an MRW and a PRW. After surgically inducing middle cerebral artery occlusion (MCAo), modified neurological severity scores (mNSS) and an inclined plane test were conducted to assess the neurological damages to the rats. PRW is experimentally validated as the most effective among such animal mobility systems. Furthermore, an exercise effectiveness measure, based on rat position analysis, showed that there is a high negative correlation between the effective exercise and the infarct volume, and can be employed to quantify a rat training in any type of brain damage reduction&#160;experiments.

This study presents an innovative running wheel-based animal mobility system to quantify an effective exercise activity in rats. A rat-friendly testbed is built, using a predefined adaptive acceleration curve, and a high correlation between the effective exercise rate and the infarct volume suggests the protocol&#39;s potential for stroke prevention experiments.

This study presents an animal mobility system, equipped with a positioning running wheel (PRW), as a way to quantify the efficacy of an exercise activity ...

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and validated for rat use. The Radial Water Tread (RWT) maze test of spatial memory (designed specifically for mice and requiring no swimming) has been shown previously to successfully distinguish between controlled cortical impact-induced TBI mice and sham controls. Here, a detailed protocol for this task is presented. The RWT maze capitalizes on the natural tendency of mice to avoid open areas in favor of hugging the sides of an apparatus (thigmotaxis). The walls of the maze are lined with nine escape holes placed above the floor of the apparatus, and mice are trained to use visual cues to locate the escape hole that leads out of the maze. The maze is filled with an inch of cold water, sufficient to motivate escape but not deep enough to require that the mouse swim. The acquisition period takes only four training days, with a test of memory retention on day five and a long-term memory test on day 12. The results reported here suggest that the RWT maze is a feasible alternative to rat-validated, swimming-based cognitive tests in the assessment of spatial memory deficits in mouse models of TBI.

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham controls.

Despite the recent increase in use of mouse models in scientific research, researchers continue to use cognitive tasks that were originally designed and ...

Assessing Activity-based Anorexia in Mice

Rodents develop activity-based anorexia (ABA) when exposed to a restricted feeding schedule and allowed free access to a running wheel. These conditions lead to a life-threatening reduction in body weight. However, rodents exposed to only one of these conditions ultimately adapt to re-establish normal body weight. Although increased running coupled with reduction in voluntary food intake appear paradoxical under ABA conditions, ABA behavior is observed across numerous mammalian species. 
The ABA paradigm provides an animal model for anorexia nervosa (AN), an eating disorder with severe dysregulation of appetite-behavior. Subjects are singly housed with free access to a running wheel. Each day, the subject is offered food for a limited amount of time. During the course of the experiment, a subject's body weight decreases from high activity and low caloric intake. The duration of the study varies based on how long food is offered daily, the type of food offered, the strain of mouse, if drugs are being tested, and environmental factors.
A lack of effective pharmacological treatments for AN patients, their low quality of life, high cost of treatment, and their high mortality rate indicate the urgency to further research AN. We provide a basic outline for performing ABA experiments with mice, offering a method to investigate AN-like behavior in order to develop novel therapies. This protocol is optimized for use in Balb/cJ mice, but can easily be manipulated for other strains, providing great flexibility in working with different questions, especially related to genetic factors of ABA.

Mice individually housed with a running wheel while given limited access to food develop reductions in food consumption and increase activity on the running wheel. This experimental phenomenon is called activity-based anorexia. This paradigm provides an experimental tool for studying the neurobiology and behaviors underlying aspects of anorexia nervosa.

Rodents develop activity-based anorexia (ABA) when exposed to a restricted feeding schedule and allowed free access to a running wheel. These conditions ...

Tracking Individual Running Metrics in Mice Using a Voluntary Wheel Running Protocol that Minimizes Social Isolation

Voluntary wheel running (VWR) in rodents is used to evaluate performance and endurance and to simulate exercise. In contrast to other exercise paradigms that require aversive stimulus to induce active movement, wheel running is voluntary. VWR generally consists of placing a running wheel inside the mouse home cage. However, analyzing exercise parameters can typically only be conducted when mice are individually housed, while overall cage effects can be reported when mice are group-housed. Therefore, the current models force experimenters to choose between measuring individual exercise parameters in single-housed mice or measuring group effects in group-housed mice; depending on the research question and model, this could be limiting. 
Therefore, the goal of this method is to demonstrate a protocol that assesses VWR in a mouse model by quantifying running parameters in individual mice, with minimal isolation. We report on device design, preoperative preparations, and experiment setup. Results of a 6-week training protocol using this VWR model suggest it provides sufficient stimulus to increase spatial object recognition memory, comparable to the clinical effect of exercise. This simple and inexpensive model facilitates investigation into the effect of VWR on multiple measurements of wheel activity, minimizing factors that could limit or affect experimental outcomes.

This protocol describes an effective voluntary wheel-running model that can be implemented in various research contexts. Unlike other voluntary running models, this method avoids isolation housing while tracking running metrics in individual mice.

Voluntary wheel running (VWR) in rodents is used to evaluate performance and endurance and to simulate exercise. In contrast to other exercise paradigms ...

Motor and Hippocampal Dependent Spatial Learning and Reference Memory Assessment in a Transgenic Rat Model of Alzheimer's Disease with Stroke

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk factor for AD, stroke has also been implicated as a risk factor and an exacerbating factor. The co-morbidity of stroke and AD results in worsened stroke-related motor control and AD-related cognitive deficits when compared to each condition alone. To model the combined condition of stroke and AD, a novel transgenic rat model of AD, with a mutated form of amyloid precursor protein (a key protein involved in the development of AD) incorporated into its DNA, is given a small unilateral striatal stroke.
For a model with the combination of both stroke and AD, behavioral tests that assess stroke-related motor control, locomotion and AD-related cognitive function must be implemented. The cylinder task involves a cost-efficient, multipurpose apparatus that assesses spontaneous forelimb motor use. In this task, a rat is placed in a cylindrical apparatus, where the rat will spontaneously rear and contact the wall of the cylinder with its forelimbs. These contacts are considered forelimb motor use and quantified during video analysis after testing. Another cost-efficient motor task implemented is the beam-walk task, which assesses forelimb control, hindlimb control and locomotion. This task involves a rat walking across a wooden beam allowing for the assessment of limb motor control through analysis of forelimb slips, hindlimb slips and falls. Assessment of learning and memory is completed with Morris water maze for this behavioral paradigm. The protocol starts with spatial learning, whereby the rat locates a stationary hidden platform. After spatial learning, the platform is removed and both short-term and long-term spatial reference memory is assessed. All three of these tasks are sensitive to behavioral differences and completed within 28 days for this model, making this paradigm time-efficient and cost-efficient.

To investigate the co-morbid Alzheimer&#39;s disease (AD) and stroke condition in a novel model, three behavior tasks are described that assess both motor control and cognitive behaviors. These tasks include the beam-walk task, cylinder task and Morris water maze.

Alzheimer&#39;s disease (AD) is a debilitating neurodegenerative disease that results in neurodegeneration and memory loss. While age is a major risk ...

Medicine

Activity-based Training on a Treadmill with Spinal Cord Injured Wistar Rats

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) on a treadmill is widely used as a rehabilitation tool in the SCI population with many benefits and improvements to daily life. We utilize this method of activity-based task-specific training (ABT) in rodents after SCI to both elucidate the mechanisms behind such improvements and to enhance and improve upon existing clinical rehabilitation protocols. Our current goal is to determine the mechanisms underlying ABT-induced improvements in urinary, bowel, and sexual function in SCI rats after a moderate to severe level of contusion. After securing each individual animal in a custom-made adjustable vest, they are secured to a versatile body weight support mechanism, lowered to a modified three-lane treadmill and assisted in step-training for 58 minutes, once a day for 10 weeks. This setup allows for the training of both quadrupedal and forelimb-only animals, alongside two different non-trained groups. Quadrupedal-trained animals with body weight support are aided by a technician present to assist in stepping with proper hind limb placement as necessary, while forelimb-only trained animals are raised at the caudal end to ensure no hind limb contact with the treadmill and no weight-bearing. One non-trained SCI group of animals is placed in a harness and rests next to the treadmill, while the other control SCI group remains in its home cage in the training room nearby. This paradigm allows for the training of multiple SCI animals at once, thus making it more time-efficient in addition to ensuring that our pre-clinical animal model mimics the clinical representation as close as possible, particularly with respect to the body weight support with manual assistance.

This protocol demonstrates our model of activity-based locomotor treadmill training for rats with spinal cord injury (SCI). Included is both quadrupedal and forelimb-only groups, in addition to two distinct types of non-trained control groups. Investigators are able to assess training effects on SCI rats using this protocol.

Spinal cord injury (SCI) results in lasting deficits that include both mobility and a multitude of autonomic-related dysfunctions. Locomotor training (LT) ...

Neuroscience

WheelCon: A Wheel Control-Based Gaming Platform for Studying Human Sensorimotor Control

Feedback control theory has been extensively implemented to theoretically model human sensorimotor control. However, experimental platforms capable of manipulating important components of multiple feedback loops lack development. This paper describes WheelCon, an open-source platform aimed at resolving such insufficiencies. Using only a computer, a standard display, and inexpensive gaming steering wheel equipped with a force feedback motor, WheelCon safely simulates the canonical sensorimotor task of riding a mountain bike down a steep, twisting, bumpy trail. The platform provides flexibility, as will be demonstrated in the demos provided, so that researchers may manipulate the disturbances, delay, and quantization (data rate) in the layered feedback loops, including a high-level advanced plan layer and a low-level delayed reflex layer. In this paper, we illustrate WheelCon&#39;s graphical user interface (GUI), the input and output of existing demos, and how to design new games. In addition, we present the basic feedback model and the experimental results from the demo games, which align well with the model&#39;s prediction. The WheelCon platform can be downloaded at https://github.com/Doyle-Lab/WheelCon. In short, the platform is featured to be cheap, simple to use, and flexible to program for effective sensorimotor neuroscience research and control engineering education.

WheelCon is a novel, free and open-source platform to design video games that noninvasively simulates mountain biking down a steep, twisting, bumpy trail. It contains components presenting in human sensorimotor control (delay, quantization, noise, disturbance, and multiple feedback loops) and allows researchers to study the layered architecture in sensorimotor control.

Feedback control theory has been extensively implemented to theoretically model human sensorimotor control. However, experimental platforms capable of ...

Whole-Brain 3D Activation and Functional Connectivity Mapping in Mice using Transcranial Functional Ultrasound Imaging

Functional ultrasound (fUS) imaging is a novel brain imaging modality that relies on the high-sensitivity measure of the cerebral blood volume achieved by ultrafast doppler angiography. As brain perfusion is strongly linked to local neuronal activity, this technique allows the whole-brain 3D mapping of task-induced regional activation as well as resting-state functional connectivity, non-invasively, with unmatched spatio-temporal resolution and operational simplicity. In comparison with fMRI (functional magnetic resonance imaging), a main advantage of fUS imaging consists in enabling a complete compatibility with awake and behaving animal experiments. Moreover, fMRI brain mapping in mice, the most used preclinical model in Neuroscience, remains technically challenging due to the small size of the brain and the difficulty to maintain stable physiological conditions. Here we present a simple, reliable and robust protocol for whole-brain fUS imaging in anesthetized and awake mice using an off-the-shelf commercial fUS system with a motorized linear transducer, yielding significant cortical activation following sensory stimulation as well as reproducible 3D functional connectivity pattern for network identification.

This protocol describes the quantification of volumetric cerebral hemodynamic variations in the mouse brain using functional ultrasound (fUS). Procedures for 3D functional activation map following sensory stimulation as well as resting-state functional connectivity are provided as illustrative examples, in anesthetized and awake mice.

Functional ultrasound (fUS) imaging is a novel brain imaging modality that relies on the high-sensitivity measure of the cerebral blood volume achieved by ...

Multifactorial Assessment of Motor Behavior in Rats after Unilateral Sciatic Nerve Crush Injury

The induction of a peripheral nerve injury is a widely used method in neuroscience for the assessment of repair and pain mechanisms among others. In addition, in the research field of movement disorders, sciatic crush injury has been employed to trigger a dystonia-like phenotype in genetically predisposed DYT-TOR1A rodent models of dystonia. To achieve consistent, reproducible and comparable results after a sciatic nerve crush injury, a standardized method for inducing the nerve crush is essential, in addition to a standardized phenotypical characterization. Attention must be paid not only to the specific assortment of behavioral tests, but also to the technical requirements, the correct execution and consecutive data analysis. This protocol describes in detail how to perform a sciatic nerve crush injury and provides a behavioral test battery for the assessment of motor deficits in rats that includes the open field test, the CatWalk XT gait analysis, the beam walking task, and the ladder rung walking task.

We provide a protocol for the assessment of motor behavior via a behavioral test battery in rats after sciatic nerve crush injury.

The induction of a peripheral nerve injury is a widely used method in neuroscience for the assessment of repair and pain mechanisms among others. In ...