The overall goal of this battery of behavioral testing is to observe the general locomotion, muscular strength, and coordination in mice. These methods can help answer key questions about the implementation of different treatments in multiple preclinical trials, such as the benefit of treatment outcomes on motility in mice. The main advantage of this technique is that multiple facets and dimensions of movement are measured and assessed by this battery of behavioral testing.
Demonstrating the procedure will be Meera, a master student in our lab. Begin by turning on the open field test program, and connecting the computer to the camera or open field box. Start the selective SmartWare software in the Open Field icon.
Select okay. If applicable, choose the Digital Analog Converter source from the camera options. Follow the instructions to make the appropriate dimensions to cover the open field box.
Next, make sure that the mouse is being detected within the confines of the open field by calibrating the appropriate part of the software. Click time to preset an amount of time to test the mice. In this case, set five minutes.
Enter the subjects, by clicking and writing the appropriate name for the files. For example, phase one, session one. Next, place a mouse in the center of the open fields, and click the start icon to prompt the program to begin recording.
After five minutes, observe the program automatically move to the next subject. In this case, remove the mouse that has completed the test and place the next subject into the open field of the program. Then click start.
Finally, to analyze the data click on the analyze icon at the top right of the screen. Click analyze again, and then summary report. Begin by placing a mouse in the center of a mesh.
And invert the mesh at lease 20 centimeters above a clear container. Once inverted, start the timer. Record the time of latency for the mouse to fall off the mesh.
If the mouse does not fall within 60 seconds, give it a score of 60 seconds, and conduct no further trials. Then, allow the mouse to rest for at least five minutes. Next turn on the rotarod and the computer.
And ensure that they are properly connected for data recording. In order to run the accelerated protocol, use codes on the keypad to set the desired length of time on the rotarod to 300 seconds, the number of lanes to five, the starting and end speed to four and forty rotations per minute, respectively, and the ramp time to 300 seconds. Then set the unit to mouse forward mode.
Ensure that all magnetized lanes are balanced before beginning the trial, and place one mouse in each lane of the rotarod. Next press start to initiate the program. As the mice fall, remove them from their lanes promptly to ensure that they do not trip the other lanes.
Finally, allow the mice to rest for a 10 minute intertrial interval. During this interval, begin the trial for the next set of mice. This data demonstrates the use of the rotarod as a means to see how the treated mice are behaving compared to the normal heterozygous mice.
The divergence of the two groups at 40 weeks denotes the beginning of the treatment group's decreased coordination in comparison to normal controls. Over monthly behavioral testing the treated mice performed similarly to the normal heterozygous controls, the open field test, rotarod, and mesh test. At 12 weeks, treated animals outperform untreated animals on time moving in the open field test and on the latency to fall in the mesh test.
Once mastered, this technique can be completed in two and half hours to complete 20 mice through the protocol, if performed properly. While attempting this procedure, it's important to remember to clean the apparatus between each mouse's trial, in order to avoid possible contaminates across mice and cages. Following this procedure, other methods such as further behavioral and biochemical assays can be performed in order to answer additional questions concerning treatment efficiency, such as biodistribution assays, enzyme activity assays, histology assays, or memory and learning tests.
After it's development, this technique paved the way for researchers in the fields of psychology and later in neurosciences to explore the affect of a variety of treatments and conditions have on model organisms. After watching the video, you should have a good understanding of how to perform behavior testing for general locomotion muscular strength and coordination. Don't forget that working with mice can be hazardous, and precautions such as wearing proper personal protective equipment should always be taken while performing this procedure.
