Activity Testing Protocol

(Final)

Apparatus and Room Setup

1. Activity monitors are enclosed in a sound attenuating chamber with a ventilation fan on. Fan provides some masking noise. Fan is a common 3" or 4" muffin fan (110 VAC). We can try to define the actual sound level, but matching may not be easy since the Portland and Albany enclosures are already in place. The Albany boxes require sliding the monitors in and out so that the top of the test chamber can be accessed. Thus the chamber is moved into place with the mouse inside it and the door then closed before the switch is thrown for session initiation.

2. Omnitech monitors should have acrylic tops on during testing since some motivated mice can actually emerge from the top. With these tops in place, additional sound dampening is available, diminishing the impact of the fans.

3. Testing is done in the dark. Room light normal if possible.

4. Computer control should be set up so that each squad's information is set up immediately following the completion of the prior squad. This permits direct movement of the succeeding squad from the colony room into the test room and into the monitors without delay for computer setup.

General Procedures

First test session begins at 8:30 am.

Second test session begins at 12:30pm.

1. Mice from the am or pm session are transported from home colony room to the holding room/hallway, adjacent to the testing room, at the beginning of the session (16 cages at a time for half the mice). Mice will stay in their home cages in this area with food/H2O available at all times. Lighting is normal room lighting.

2. Mice are tailmarked before testing begins. Another experimenter can tailmark while one starts the activity testing. Black mice will be tailmarked with a red sharpie pen while all others are tailmarked with a black sharpie pen.

3. Transport into the testing room is done two cages at a time (four mice, one squad), in the home cage (bottle removed, except for Portland - see common procedures protocol for explanation).

4. Monitors are cleaned with dilute isopropyl alcohol wipes at the end of each squad's test. A first wipe with a paper towel to remove feces/urine is helpful prior to the wipedown with isopropyl. Doing this at the completion of each squad give some time for odor to dissipate prior the acquisition and test initiation for the next squad.

5. Handling is done with gloved hands at all times (for this procedure and all others).

6. A 15 min test with three five min blocks will be conducted. The 15 min length permits roughly three squads per hour (12 mice).

Specific Protocol for Testing

1. Mice are tested in squads of four. This involves two cages of littermate pairs each. Each squad is transported by hand to the testing room where activity monitors are all set to receive them.

2. Put one cage on the towel on the table. Remove the lid from the other cage and place it gently on the towel. Then carry the open cage to the monitors.

3. Grasp mouse by the tail. Place it in the center facing away from you. Replace the lid, turn on the monitor switch, and close the sound attentuation chamber. Repeat for all four mice.

4. Tests are 15 min tests, set up with three five min blocks.

5. As each mouse's session completes, the mouse can be removed. At the completion of each mouse's test, open the chamber, switch off the control for the box, remove the mouse and replace it in the appropriate waiting home cage. After all four, the control program is tended so that the files are saved, and the next squad set up.

6. Count boli in each monitor and record these data, plus any other noteworthy observations. All monitors are then cleaned with 10% isopropyl alcohol and Kimwipes. A first wipe with a paper towel to remove feces and excess urine is helpful prior to wipedown with isopropyl. Leave the plastic lids off to allow any fumes to dissipate prior to the next squad.

7. Mice are transported back to the holding room. Carry the two cages back tot he holding room and bring in the next squad. At the end of the AM or PM session, the set of 16 cages are moved back to their colony room position.