Strategies for Assessing Autistic-Like Behaviors in Mice

Our research focuses on the pathophysiology of autism spectrum disorder, ASD. And specifically, we look at the molecular mechanisms involved in ASD and what causes the behavioral impairments. These together will involve specifical treatments related to ASD and will improve the life of individuals suffering from ASD.

One of the challenges in our research is to improve research with ASD-related behavior and communicate that between different laboratories. There's a lot of variation between the different laboratories, how they do the behavioral assessments, and that is a big problem to advance treatments into the field. So we hope with our protocols, we will help these advancement and the communication between laboratories around the world.

By optimizing and detailing the grooming and the three-chamber test protocols, we establish a consistent framework that researchers can rely on to obtain reliable and reproducible results from ASD mouse models. This standardization will facilitate better cooperation and interpretation of findings from different laboratories, leading to a better understanding of ASD and the development of potential therapies. Our laboratory is interested in explaining cell-type-specific translation process in autism-like behavior.

Additionally, we also studied the potential therapeutic applications in ASD mouse models, such as metformin. To begin, switch on dimmed lights. Clean the table and set up the room.

Prepare each test cage with a thin layer of fresh bedding. Place one to two cages on the table, separated from each other and the room environment by white plastic blinders. Then, place a camera before each cage to capture the test mouse.

Cover the cages with a fabric sheet, and transfer all mice into the room to avoid stress during the transfer process. Once the mice are placed in the testing room, remove the sheet. Before the experiment starts, leave them in the room with dimmed lights for at least 30 minutes.

At the start of the test, write down the mouse identification number and testing information on a whiteboard. Place the test mouse into the cage with fresh bedding. Start recording and place the test mouse into the test cage.

Leave the room during the 20-minute recording session. After 20 minutes, return to the room. Present the whiteboard to the camera and stop recording.

Place the test mouse back into the home cage and repeat the experiment. After the experiments, return the mice to their housing room. The self-grooming test revealed increased grooming time in Eif4ebp2 knockout mice compared to control mice.

Eif4ebp2 knockout mice exhibited significantly more grooming bouts than control mice. To begin, clean the table, and turn on the dimmed lights. Place the three-chamber plexiglass instrument on the table surrounded by privacy blinds.

Place one camera overhead, ensuring that all three chambers are in the recording frame of the camera. Then, transfer the test and stranger mice into the room, covering the cages with a fabric sheet during the transfer process. Remove the sheet once they are in the room.

Leave the mice in the room for at least 30 minutes with dimmed lighting before the start of the experiment. Keep the three chambers empty during habituation. After selecting a test mouse, write the identification number on the whiteboard.

Place the test mouse in the center compartment while the sliding doors remain closed. Open the doors and start recording to allow the test mouse to explore the three empty chambers for 10 minutes. After 10 minutes, return to the room.

Gently guide the test mouse to the center compartment and close the sliding doors. Display the whiteboard and stop the camera. Position two wire cages in the left and right chambers diagonally opposite each other in the corners of each chamber.

Measure the distance between the cages and the chamber walls. Ensure the cages do not directly face the chamber doors. Choose one stranger mouse and introduce it into one of the wire cages, leaving the other cage empty.

To start the test, press record and remove both sliding doors before leaving the room. Allow the test mouse to explore an empty wire cage in one chamber and a cage containing a stranger mouse in the other chamber for 10 minutes. After 10 minutes, return to the room.

Display the whiteboard to the camera and end the recording. Reintroduce the mouse to the center compartment and close the interconnecting doors. Place a new, never-encountered mouse in the previously empty wire cage.

Start recording and open the interconnecting doors to allow the test mouse to explore the apparatus for 10 minutes before leaving the room. The test mouse will explore two wire cages, one with the previously encountered S1 mouse and the other with the newly introduced S2 mouse, for 10 minutes. After 10 minutes, return to the room.

Display the whiteboard to the camera and conclude the recording. Return all the mice to their home cages. Thoroughly clean the chambers and wire cages using an odor-free disinfectant, and dry the apparatus before the next experiment.

After testing, return the mice to the housing facility. Mice preferred interacting with S1 over a novel inanimate object. Similarly, C57BL/6J mice displayed more interest in a newly introduced S2 than a familiar S1.