Mouse motor performance was evaluated weekly using a rotarod apparatus (Muromachi Kikai, Tokyo, Japan), as described previously [17 (link)]. After the training period of 14 days, mice were able to stay on the rotarod rotating at a speed of 24 revolutions per minute (rpm). The maximum allowable score was 300 s, and the average time of three trials for each mouse was recorded twice a week. The observers were blinded with regard to treatment by MBP but performed their assessment concurrently. The end-point was defined as the inability of the mouse to right itself within 30 s after being placed on its side [2 (link)]. At that point, mice were euthanatised with CO2.
Rotarod apparatus
Manufactured by Muromachi Kikai
Sourced in Japan
The Rotarod apparatus is a device used to assess motor coordination and balance in laboratory animals. It consists of a rotating rod that rotates at a controlled speed, and the animal is placed on the rod to measure its ability to remain on the rotating surface. The core function of the Rotarod apparatus is to provide a standardized method for evaluating motor function in animal models.
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5 protocols using rotarod apparatus
1
Evaluating Motor Performance in Mice
2
Comprehensive Motor Function Evaluation
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Motor function was evaluated using the BMS, Rotarod apparatus (Muromachi Kikai), and DigiGait system (Mouse Specifics). For histological analyses, the animals were anesthetized and transcardially perfused with 4% paraformaldehyde (pH 7.4). Spinal cords were removed and sectioned in the sagittal/axial plane on a cryostat. All motor function and histological analyses were conducted by observers blinded to the treatment conditions. All animal experiments (approval number 09169) were performed under the control of the Keio University Institutional Animal Care and Use Committee in accordance with the Institutional Guidelines on Animal Experimentation at Keio University, the Japanese Government Law Concerning the Protection and Control of Animals, and the Japanese Government Notification of Feeding and Safekeeping of Animals, and approved by the ethics committee of Keio University (IRB approval number 09091-8).
3
Rotarod Test for Motor Coordination
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Example 4
Rotarod Test
This test is performed to evaluate the effect of the compound on the motor coordination.
The example compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 21, 22, 23, 24, 25, 26, 28, 30, 31, 37, 38, 40, 41, 42, 44, 45, 47, 48, 50, 52, 53, 54 and 56 were used as test compounds in this test.
Male ICR mice (Japan SLC, Inc., 5-6 weeks, 8 per group) were trained to remain on a fixed speed (15 rpm) rotating rod of rotarod apparatus (Muromachi Kikai Co., Ltd.) for 2 minutes. The test compound was suspended in 5% gum arabic/distilled water (w/v), and administered by forced oral administration at a dose of 30 mg/kg. After 1 hour of oral administration, the mice were again placed on the rod accelerated from 4 rpm to 40 rpm over 5 minutes and the latency to fall off the rod was recorded for 200 seconds. The falling latency of the compound administration group was calculated as a relative value relative to the average value of the falling latency in the solvent administration group.
In this test, the rate of motor dysfunction with the example compounds 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 21, 22, 23, 24, 25, 26, 28, 30, 31, 37, 38, 40, 41, 42, 44, 47, 48, 50, 52, 53, 54 and 56 was 25% or less.
4
Evaluating Motor Coordination in Mice
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Motor incoordination is one of the most common AEs associated with the use of ASMs.9 Therefore, motor coordination was evaluated in mice receiving E2730 to assess if it is a new ASM candidate with a wide margin between therapeutic effects and AEs. E2730 (50–400 mg/kg) or vehicle was orally administered in ddY mice (male, 5 weeks old) 1 hour before the rotarod test, and the latency for mice to fall off the rod was measured. The rod rotation of the rotarod apparatus (Muromachi Kikai) was continuously accelerated from 0 to 40 rpm in a cut‐off time of 180 seconds.
5
Rotarod Test for Motor Function
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The tests using an accelerating-rotarod (72) were conducted for consecutive 3 days. On the first day, animals were habituated to the rotarod apparatus (Muromachi Kikai Co., Ltd., Tokyo, Japan) at constant 4 rpm for 30 s. Four trials on the rotarod linearly accelerating from 4 to 40 rpm in 300 s were performed in a day with a 600 s interval. The latency when the mouse fell off was recorded.
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