Service Center for Neurobehavior

The Rodent Behavioral Center in the Department of Neuroscience at UConn Health has recently upgraded many apparatuses with the latest version of ANYMAZE software to acquire animal behavioral data. The center is a collaborative partner with PIs interested in determining the functional phenotype of their experimental animal models. The facility will provide a unique opportunity for all research laboratories at UConn Health for training their lab personnel to perform animal behavioral tests. Our goal is to provide a convenient resource for in-depth characterization and analysis of their animal models, making them more effective research tools for future studies and grant applications.

In order to use the center, scientists should have the behavioral procedures in their approved IACUC protocol. Following professional training, scientists will be allowed to book and access the behavioral rooms which are located in CCM Tower on the 4th floor.

Behavioral Assays

Morris water maze

This is the most widely used test for measuring spatial navigation and reference memory. The test animal is placed in an open, circular pool in which there is a submerged platform. A series of tests can measure a variety of memory and navigation tasks, as the test animal swims to the platform from various locations, or tries to find the platform after it has been removed. Tests are video-tracked using AnyMaze, and analysis can provide information on the test animal’s hippocampus-dependent reference memory and prefrontal cortex working memory.

Purpose/Goal:

  • This test is used to assess spatial memory. Paradigms that evaluate reference and working memory are available.
  • Primary dependent variables of interest include latency, distance, and proximity to goal platform.
  • Total swim distance and average speed can be calculated to determine if motor deficits interfere with cognitive function.
  • The visible platform test ensures visual impairment does not interfere with cognitive performance.

Timeline:

Paradigm Number of consecutive days
Handling 1
Visible platform 3
Reference memory/Probe trial 5
Working memory 3

 

Fear conditioning

This test uses cued conditioning to evaluate amygdala function and contextual conditioning to evaluate hippocampal function. Animal is placed in a conditioning chamber (Med Associates) for 3 minutes before onset of a 30 seconds conditioning sound stimulus (CS, 2800Hz&85dB). An unconditioned stimulus (US) of continuous foot shock (0.5mA) occurs during the last 2 seconds of the CS. After resting an additional 30 seconds in the chamber, the animal is repeated the CS and US again, and the animal remains in the chamber for an additional 30 seconds before being returned to its home cage. This environment provides reinforcement for contextual and cued conditioning. To evaluate contextual conditioning, the animal is placed in the same chamber without the CS and US for 3 minutes to assess its freezing behavior. To evaluate cued conditioning, the animal is placed in a novel chamber with a smooth flat floor, different colored walls, and a novel odorant. The animal remains in the chamber for 3 minutes before onset of only a 30 seconds CS. No US will be delivered. After resting an additional 30 seconds in the chamber, the animal is repeated the CS again, and return to its home cage after 30 seconds resting in the chamber.

Purpose/Goal:

  • This test is used to measure learning and memory.
  • Freezing behavior during testing is the primary dependent variable of interest.
  • This test can be run multiple times over several weeks/months to evaluate effectiveness of conditioning.

Timeline:

Paradigm Number of days Protocol
Handling 1  
Conditioning 1  
Contextual recognition 1  
Cured recognition 1  

 

Y-maze

This test is commonly used to evaluate animal spatial working memory. The maze is in the shape of a “Y” with three equally sized arms. An animal is placed in the center of the Y maze and given 5 minutes to freely explore. There is no pre-training involved in this task and animals experience very little distress. The animals’ movement is recorded with a video camera and AnyMaze software.

Purpose/Goal:

  • This test specifically targets the hippocampus, the portion of the brain important in spatial orientation and navigation.
  • Primary dependent variables of interest include total number of arms entered, spontaneous alteration, and number of repeated arms entered.
  • Higher percentages of spontaneous alteration reflect higher functioning spatial working memory

Open field

This test is used to measure locomotor activity in rodents. It is a test based on natural exploratory instincts in a novel dark open field. Animals are placed in an open field arena for 10 minutes and their free movement is recorded via AnyMaze software. Exploratory activity, anxiety, and locomotion can be evaluated by analyzing total moving distance, average speed, center time & distance, corner time & distance, etc.

Purpose/Goal:

  • The open field test is used to evaluate locomotor activity.
  • This test can be useful when studying central and peripheral pathways involved in locomotion, anxiety, and exploration.
  • This test can be run multiple times over several weeks/months to document chronic changes in activity.

Novel object recognition

This test uses the animal’s reaction to a novel object as an avenue to evaluate dorsal hippocampal function. Following habituation, the animal is placed in an open field with two of the same objects and given a specific period of time to freely explore before being removed from the testing environment. After a specific time delay, the animal is then placed in an open field with two different objects, one of which is the previous familiar object, and one of which is an unrecognized novel object. For each trial, the time the animal spends actively exploring (sniffing, touching, climbing, etc.) each object is recorded using AnyMaze software.

Purpose/Goal:

  • Novel object recognition is used to evaluate recognition memory.
  • This test can be useful in determining whether mice prefer to spend more time with novel object.

 

Timeline:

Paradigm Number of consecutive days
Handling 1
Habituation 1
Exposure to 2 identical objects 1
Exposure to 1 familiar, 1 novel object 1

 

Social Interaction

This test is used to assess animal cognition in the form of general sociability and interest in social novelty. Rodents normally prefer to spend more time with another rodent (sociability) and will investigate a novel intruder more so than a familiar one (social novelty). Testing occurs in three sessions within a three-chambered box with openings between the chambers. After habituation to the empty box, the subject encounters a never-before-met intruder under one pencil cup and an empty pencil cup in the “sociability” session. The subject then encounters the first intruder as well as a second never-before-met intruder under another pencil cup in the “social novelty” session. The time spent sniffing each pencil cup, the time spent in each chamber, and the number of entries into each chamber are presented. This test is useful for quantifying deficits in social behavior in transgenic animals exhibiting autistic traits as well as evaluating novel chemical entities for their effect on social behavior.

Purpose/Goal:

  • This test can help identify rodents with deficits in sociability and/or social novelty.
  • This test can be useful in determining whether animals have neuropsychiatric disorders, such as depression, autism, schizophrenia, et al.

Timeline:

Paradigm Number of consecutive days
Handling 1
Habituation 10 min in Day2
Sociability 10 min in Day2
Social Novelty 10 min in Day 2

 

Barnes Maze

Barnes maze training and testing is performed using a 20-hole white Barnes table (San Diego Instruments, San Diego, CA, USA). The paradigm is a composite of several published methods, all emphasizing not to overtrain mice and to test long-term memory after a several day interval (hiatus) between the final training trial and the probe or test day. Mice are habituated on Day 0 to the platform for 2 min and then gently guided to the escape compartment with the handler’s hand. After 1 min in the escape compartment, the mouse is returned to the home cage in the escape compartment. The computer has no password, just hit “Enter”. On Days 1, 2 and 3, each mouse has 2 training trials of 3 min spaced by 15-25 min; trials are initiated when an opaque beaker is raised from over the mouse. If the mouse does not reach the escape hole in 180 sec in any training trial, it is gently guided to the hole. Once in the escape compartment, the mouse is kept there for a min and then is carried back to the home cage in the escape compartment. After a 72h respite, the mouse is tested for 1 additional probe trial. Nose pokes are recorded for all holes by the computer; data reported include the percentage of nose pokes in the proximal quadrant (“A”; i.e. the quadrant with the escape hole), the primary latency (time to first poke into the escape hole, regardless of whether the mouse enters the escape compartment or not), and the number of “mistakes” (nose pokes in quadrants B+C+D).

Rotarod

Rotarod testing is performed three times a day for 3 days, using a Five Lane Rota-Rod for Mouse (Med Associates, Georgia, VT). Each trial lasts no longer than 5 min, with the speed increasing from 4 rpm to 40 rpm, or until the mouse falls off. The longest time recorded for each day is used in the analysis. Care must be exercised to ensure the mice do not cross into other lanes after falling off, to preserve mouse identity and to avoid incorrect registration of a beam break (fall).

Scheduling the rooms

Card access, along with initial training, and signup are required to enter these suites. The equipment and calendars for each room are designated below. Please review room availability first. If available, please email servicecenterforneurobehavior@uchc.edu to make your reservation(s).

 

 

Need badge access? To request badge access to these rooms, please email Jody Gridley, gridley@uchc.edu.

Contact Us

Xiangyou Hu, Ph.D.

Assistant Professor

Department of Neuroscience

UConn Health

Phone: 860-679-6249

Email: xhu@uchc.edu