Mouse Behavior Laboratory (MBL)
The Mouse Behavior Laboratory
Our mouse behavior laboratory includes a dedicated housing room (263 sq ft) for up to 160 cages of mice in a ventilated rack in the T-wing of the Health Sciences Building, and three large procedure rooms (762 sq ft) subdivided into seven separate testing areas. The facility is located within a centralized, specific pathogen free Vivarium (approx. 10,000 sq ft) in the T-wing of the Health Sciences Center, which provides husbandry and veterinary support, caging, supplies, and cage-wash facilities.
Steps for Initiating a New Project
For projects in the proposal development phase, as well as for approved projects, the MBL works with affiliates to provide scientific, management, and budgetary information. Funded grants also are reviewed to ensure strict compliance with IACUC standards, careful observance of the NIH Guide to the Care and Use of Laboratory Animals, and compliance with accreditation standards set by AAALAC. For funded projects, access to Core services that are required for individual research projects is discussed with the investigators. Of note, all services described above are provided at no cost (e.g., training, consultation, coordination, testing equipment). Construction of highly specialized equipment, however, may require investigator support. The Department of Comparative Medicine does charge affiliates the decentralized per diem rate for housing mice in our testing facility.
Use of facilities is scheduled on a daily basis through an online calendar. Laboratory schedules that list the day-to-day research activity in the labs are also posted in the testing rooms.
Please contact Toby B. Cole, Ph.D., email@example.com, 206-543-9300.
Mouse Behavior Laboratory Facility 206-221-7283
Behavioral Testing Capabilities
Note - behavioral testing typically requires a minimum of 10 mice per experimental group. Testing is highly sensitive to environmental influences, so steps to minimize environmental differences are critical. Tests available in the MBL include a large variety of cognitive and neuromotor behavioral assessments. Neurobehavioral domains capable of being assessed include activity/exploration, motor function, learning and memory, anxiety, sensory functions, neurological reflexes, and social behavior.
SR-Lab Startle Response System
The SR-LAB System from San Diego Instruments utilizes auditory or air puff startle to test the startle habituation and pre-pulse inhibition paradigm. SR-LAB software allows researchers to develop and execute any startle reflex testing paradigm including customized paradigms.
The rota-rod treadmills from Med Associates are designed to accurately measure motor coordination, fatigue, and cerebellar learning. Each rota-rod treadmill consists of a computer-controlled stepper motor-driven drum with constant speed or accelerating speed modes of operation. The drum has been specially machined to provide a suitable grip for the animal. For mice it is divided into five test zones so that up to five animals may be tested at the same time. When the animal falls off the rotating drum, it breaks a photobeam to automatically record the amount of time spent on the drum.
The open field arena is a 48 cm by 48 cm plexiglass square enclosure designed to measure anxiety and exploratory drive. The amount of movement and the percent time spent in the center of the open field are measured. Mice typically exhibit thigmotaxis, the tendency to remain close to the walls of the arena, but with decreased anxiety or increased exploratory drive will explore the center area more extensively.
Morris Water Maze
The custom built water maze consists of a pool, with a hidden platform submerged just below the water surface. It is used to assess behaviors including spatial learning, memory, reversal, and delayed-match-to-sample.
8-arm Radial Maze (water maze)
The custom built 8-arm radial maze is used to assess spatial learning, memory, and working memory and can be configured for water Y-maze and water T-maze assessments.
Shuttle Box with Precision Shocker and Noise Generator
The system from Coulbourn Instruments is designed to assess passive avoidance, active avoidance, contextual and cued fear conditioning, and light/dark preference. There are three bays on each side of the cage to accept a wide variety of stimulus and response modules. A shock floor is wired for eight unique shock outputs.
HABITEST Modular Test Cage Systems
The HABITEST system from Coulbourn Instruments is designed to assess contextual and cued fear conditioning and operant conditioning. The system has multiple configurations including the use of isolation boxes, shock floors, pellet dispensers and cue lights. Graphic State Notation software supports the HABITEST system. The software requires no programming skills and has pre-built protocols for ease of use. The system can test multiple subjects simultaneously using either the same protocol or up to 16 different ones. An automated protocol check procedure ensures that protocols are complete and will terminate properly. Data analysis is provided in both numeric and graphic format and includes selectable analysis structures for both single subject and group averages.
This equipment assesses footfalls and gait quantitatively. It consists of a corridor that directs and limits the freedom of movement of the mouse to a straight line across the walkway, where a special glass walking surface and high speed color camera record and analyze the gait. The mouse is placed at one end of the corridor, and walks to the other end, then is returned to its home cage. For motivation, it is useful to include a goal box at the end, consisting of a black shelter with room for a home cage underneath, or to provide an appetizing edible treat.
Activity Monitoring in Noldus PhenoTyper Cages
The MBL has 16 PhenoTyper cages, consisting of a basic cage or a home cage, with a top unit that is equipped with LED units and an IR camera. One or two mice are placed into the cage, and they are left in the cage for as long as the experiment requires (up to one week), while their activity is recorded with the Ethovision system. Because they have access to food and water, mice can be monitored during both the light and dark cycle, allowing determination of circadian patterns of activity. Optional running wheels and, lickometers to measure intake of liquid (for use in trials of drug administered via drinking water) have recently been added.
The PhenoWheels from Noldus are running wheels that combine with EthoVision® XT, a Trial & Hardware Control Module, and a USB-IO box to measure the activity of mice inside the running wheels. It can be placed inside any PhenoTyper basic or home cage. The activity in the running wheel is measured by means of a magnet and a magnetic sensor; the Counter Module counts the number of rotations and sends this information to the EthoVision XT software. IDDRC affiliates can use EthoVision XT to measure the amount of time spent in the running wheel. The data from the PhenoWheel Counter Module can also be used as conditions in test protocols that are executed by EthoVision XT's Trial & Hardware Control Module.
UltraVox is the professional system for the automatic detection of ultrasonic vocalizations such as mouse pup calls in neurotoxicological studies or rat vocalizations in drug testing. It visualizes realtime filtering and continuously updates descriptive statistics to offer quick insight into the temporal structure of vocalizations. The UltraVox system combines an ultrasound detector, audio filter, and the UltraVox software. It allows researchers to monitor vocalizations with any type of temporal structure. Up (ultra) sound detectors can be connected to the UltraVox system to observe the effects of different treatments. Researchers can also determine how and when data acquisition starts by using the automatic start options. Separate time-event plots for the unfiltered and filtered signals provide direct visual feedback as to the effectiveness of specific acquisition settings.
Mazes and Video Tracking Equipment
Multiple videotracking systems with cameras and EthoVision tracking software are available to researchers, two for use with desktop computers and one for use with a laptop computer. All three systems are capable of either live tracking or tracking animals in recorded video files. These systems provide a high degree of flexibility, and can be used with a large variety of mazes, including the T-maze, elevated-plus maze, Morris water maze, radial-arm maze, light-dark boxes, open field boxes, and social approach arenas.
EthoVision XT base software from Noldus allows researchers to track and analyze movement, activity, and behavior of one animal in one arena. A wizard guides researchers through all phases of the experiment, from setting up the project to detailed analysis of the data acquired. A large number of templates for all standard tests are included in the software, allowing researchers to start experiments quickly and easily. The software can reliably detect and track any kind of animal, even when there is low contrast. EthoVision XT also allows in-depth analysis for such data as: time spent immobile (serving as a measure of fear) and rotation (research on a variety of brain disorders). Researchers can select tracks or parts of tracks by the powerful data selection criteria the software offers. Selecting data in which animals receive a specific treatment allows researchers to only analyze relevant data. Data can be nested on the basis of time, zone, or behavioral state. The results can be visualized in a time-event plot together with the track data and the video file to provide an instant informal feel for research data.
EthoVision Multiple Body Point Module
The Multiple Body Point Module from Noldus tracks the nose-point and tail-base along with the center-point of a rat or mouse. Researchers can accurately measure the position of the rodent’s nose in relation to objects. This module can also be used to more accurately analyze entrance into, for example, the arms of the elevated plus maze, define which body points should have entered the arms before it is counted as an entry, or analyze feeding and drinking behavior of the animal based on the nose point being located at the position of the feeder or water bottle.
EthoVision Social Interaction Module
With the Social Interaction Module from Noldus researchers can track multiple animals per arena simultaneously and calculate a list of social parameters, such as the proximity of one animal to another or the time an animal spent moving away from another animal. In combination with the Multiple Body Points Module, nose-nose interactions and nose-tail interactions of rodents can also be analyzed.
Media Recorder is a software tool from Noldus that enables synchronous video recordings from up to four different sources. Media Recorder allows researchers to easily combine several recordings from one study. Video files may be stored separately or combined into one single file. Researchers may also choose to combine four video streams into one file when studying animal interactions from several different camera angles.
The Noldus lickometers accurately detect any contact made with the spout of a water bottle. They detect any change in capacitance between the spout and the metal plate attached to an electronic box. A change in capacitance only occurs when either a rat or a mouse makes contact with the spout. This technology differs from conventional technology where the animal becomes part of an electrical circuit that involves a current flow passing through the animal.
Social Interaction Arenas
These arenas are each comprised of a three-chambered plexiglass test box that can be used for tests of social approach, social novelty, and social discrimination, as well as novel object recognition. For social approach testing, the mouse is placed in the center compartment and given a choice between spending time in the side containing an unfamiliar (stranger) conspecific mouse, or remaining alone. The stranger mouse is contained within a small cylindrical wire cage, to allow exposure to visual, auditory, olfactory, and some tactile stimuli, while preventing aggressive or sexual interactions. Noldus tracking software is used to track the mouse’s movements and quantify time spent in each chamber and engaging in different behaviors. An identical wire cage in the opposite side chamber serves as a control. Mice normally demonstrate a clear preference for spending time in the proximity of another mouse. To test social novelty, a second unfamiliar mouse (stranger 2) is placed into the wire cage that was empty during the assessment of social approach. Mice generally show a preference for the newly-introduced stranger 2. These arenas can also be used to perform novel object recognition testing for assessment of learning and memory. Mice are habituated to the arena on the first day, and the next day, mice are presented with two novel objects for 6 minutes. Retention is then tested at various times by placing one familiar and one novel object in the arena and measuring percentage of time spent in proximity to each object.
Mice are placed on an elevated grid surface (30 L x 35 W x 31 H cm) and the number of foot-faults made by the ipsilateral and contralateral limbs are counted. Each test consists of three trials lasting 1 min each, with an inter-trial interval of 1 min. Foot faults are expressed as the number of errors made by the contralateral limbs as a percentage of the total errors made.
Mice are allowed to alternate spontaneously between the left and right goal arms (30 x 10 cm) of a T-shaped maze (start alley 30 x 10 cm) throughout a 15-trial session. Once a subject has entered a particular goal arm, a door is lowered quietly to block entry to the opposite arm. The door is re-opened after 30 sec when the mouse is returned to the start arm and allowed to choose between the goal arms. Each trial takes about 2 min. The spontaneous alternation rate is calculated as the ratio between the alternating choices and total number of choices. If necessary for motivation, "rewarded alternation" may be used, in which the mouse is made hungry by 12-hr food deprivation and is then rewarded with a preferred food if it alternates.
The test is performed identically to the non-elevated version, except that the maze is elevated 30-cm above floor level, and low walls (1 cm) made of clear plastic are used instead of high walls.
Radial Arm Maze
The land version of the 8-arm radial maze is identical to the water radial arm maze (#5 above), except it uses food as a motivator instead of water-escape.
The elevated-plus maze measures anxiety, exploration and activity levels in mice by taking advantage of the innate tendency of mice to avoid open and elevated areas. The maze consists of a central square (5 x 5 cm), from which radiate four arms (5 x 30 cm). Two of the arms have plexiglass walls (15 cm high) around the edge (closed arms), whereas the other two arms do not have walls (open arms), but do have a 0.25 cm-high edge to prevent the mice from falling from the maze. The maze is elevated 45 cm above the floor. A videotracking system is used to measure entries and duration in the center, open arms, and closed arms. The test can be performed in the light or dark cycle. Mice are placed in the central square of the apparatus, facing an open arm. Mice are allowed to explore the apparatus for 5 min while data are being collected. Data collected include line crosses, rears, head dips, grooming, stretch attend postures, urination puddles, fecal boli, closed-arm entries and duration, open-arm entries and duration, center entries and duration.
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