To design a Standard Operating Procedure that describes the procedure for the motor coordination activity using beam walking in experimental mice.
2.0 SCOPE
This SOP provides details on handling, randomization, oral dosing, use of beam walking apparatus, and evaluation of motor coordination activity in mice.
3.0 RESPONSIBILITY
Chief scientists in charge, study directors, or study investigators shall be responsible for taking the above experimental procedure.
4.0 Procedure
4.1.1 Mice will be quarantined for 1 week as per the in-house SOP if required.
4.1.2 Mice will be kept in a thermally regulated animal house (24˚C±1˚C) with a relative humidity of 50% ±20% or renewed air supply or 12 hours dark/light cycle.
4.1.3 All mice will be weighed using a digital animal weighing balance and recorded in the lab notebook.
4.1.4 Mice will be randomized based on body weight into different groups or as per the study protocol.
4.1.5 Same experimenters will conduct the whole study and avoid to use of perfume or soaps with a strong odor.
4.1.6 The animals will be handled for several days before the test (picking it up gently by the torso and holding it for a minute or two) to acclimate them to the experimenter and instrument as well.
4.1.7 The middle 80cm of each beam will be defined by drawing lines 10 cm from the beginning and 10 cm from the end of the beam using a permanent marker.
4.1.8 A “start” will be mounted at end of a beam on a narrow support stand on a stable laboratory bench in the experimental room and a goal box will be attached at the other end.
4.1.9 Two 60-W desk lamps will be positioned above and to one side of the start of the beam to create an aversive stimulus (bright light).
4.1.10 This bright light will encourage mice to traverse the beam to the dark enclosed goal box.
4.1.11 A video camera will be attached to a tripod and positioned 1.5 to 2m away from the center of the beam so that the full length of the beam fills the viewfinder of the video camera.
4.1.12 The mice will be transferred, in their home cages, from the holding room to the experimental room and allow mice to habituate to the experimental room for 60 min.
4.1.13 The lamp will be turned on so that they illuminate the start area of the beam and
recording with the video camera will be started.
4.1.14 A mouse will be placed at the start of the training beam, and the latency to traverse the beam (up to 60sec) will be recorded.
4.1.15 The mouse will be allowed for four consecutive trials and returned to its home cage.
4.1.16 The apparatus will be cleaned with 70% ethanol and allow alcohol to evaporate fully before reusing the apparatus.
4.1.17 At the end of the day’s training session, all mice will be returned to home cages and clean apparatus thoroughly with 70% ethanol.
4.1.18 The trial will be repeated for three consecutive days to generate a stable baseline response.
4.1.19 On the day of the primary test each mouse will be allowed for two consecutive trials on each of the square and round beams.
4.1.20 Mice will be tested first with the widest beam and then with the narrowest beam in each case.
4.1.21 The latency to traverse each beam and the number of times the left and right hind feet slip off each beam for each trial will be measured.
4.1.22 After completing two trials on each of the six beams, the mice will be returned to their home cages.
4.1.23 Mean scores of the two trials for each beam for each measure (i.e., latency, right hind foot slip, left hind foot slip) will be analyzed.
5.0 REFERENCES
5.1.1 Fish, F.E. 1996. Measurement of swimming kinematics in small terrestrial mammals. In Measuring Movement and Locomotion: From Invertebrates to Humans (K.P. Ossenkopp, M. Kavaliers, and P.R. Sanberg, eds.) pp. 171-188.
5.1.2 R.G. Landes, Austin, Tex. Gage, F.H., Dunnett, S.B., and Björklund, A. 1984. Spatial learning and motor deficits in aged rats. Neurobiol. Aging 5:43-48.
5.1.3 Gentile, A.M., Green, S., Neiburgs, A., Schmelzer, W., and Stein, D.G. 1978. Disruption and recovery of locomotor and manipulatory behavior following cortical lesions in rats. Behav. Biol. 22:417-455.
5.1.4 Golani, I. 1976. Homeostatic motor processes in mammalian interaction: A choreography of the display. In Perspectives in Ethology, Vol. 2 (P.P.G. Bateson and P.H. Klopfer, eds.) pp. 69-134. Plenum Press, New York.
5.1.5 Golani, I., Wolgin, D.L., and Teitelbaum, P. 1979. A proposed natural geometry of recovery from akinesia in the lateral hypothalamic rat. Brain Res. 164:237-267.
5.1.6 Goldstein, L.B. and Davis, J.N. 1990a. Beam-walking in rats: Studies towards developing an animal model of functional recovery after brain injury. J. Neurosci. Methods 31:101-107
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