Name: comprehensive understanding of inactivity-induced gait alteration in rodents
Uploaded: 2022-07-06T13:10:00
Description: Watch this Scientific Journal Video about Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents at JoVE.com

This study was supported in part by the Japan Society for the Promotion of Science (JSPS) KAKENHI (no. 18H03129, 21K19709, 21H03302, 15K10441) and the Japan Agency for Medical Research and Development (AMED) (no. 15bk0104037h0002).

The present study was approved by the Kyoto University Animal Experimental Committee (Med Kyo 14033) and performed in compliance with National Institute of Health guidelines (Guide for the Care and Use of Laboratory Animals, 8th Edition). 7-week-old male Wistar rats were used for the present study. A schematic representing the sequence of procedures is provided in Supplementary File 1.
1.&#160;Familiarizing rats with treadmill walking
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Alteration of environments leads to fluctuating functional aspects and musculoskeletal components of locomotor systems26,27. Aberrations in contractile structures or environments may affect functional abilities, persisting even after resolving mechanical/environmental distortions19. Objective motion analysis helps to measure those functional abilities quantitatively. As shown above, video analysis is a powerful methodology for acquiring su...

Lack of physical activity or disuse leads to the deterioration of locomotor effectors, such as muscle atrophy and bone loss1 and whole-body deconditioning2. Moreover, it has recently been noticed that inactivity affects not only structural aspects of musculoskeletal components but also qualitative aspects of the movement. For example, the limb positions of rats exposed to a simulated microgravity environment were different from those of intact animals even 1 month after the intervention ended3,4. Nevertheless, little has been reported on motion deficits caused by inactivity. Also, comprehensive motion characteristics of the deteriorations have not been fully determined.
The current protocol demonstrates and discusses the application of kinematic evaluation to visualize motion alterations by referring to gait motion deficits evoked through disuse in rats subjected to hindlimb unloading.
It has been shown that hyperextensions of limbs in walking after a simulated microgravity environment are observed both in humans5 and animals4,6,7,8. Therefore, for universality, we focused on general parameters in this study: angles of the knee and ankle joints and vertical distance between the metatarsophalangeal joint and hip (roughly equivalent to the height of the hip) at the middle point of the stance phase (midstance). Further, potential applications of video kinematic evaluation are suggested in the discussion.
A series of kinematic analyses may be an effective measure to assess functional aspects of neural control. However, although motion analyses have been developed from footprint observation or simple measuring on captured video9,10 to multiple camera systems11,12, universal methods and parameters are yet to be established. The method in this study is intended to provide this joint motion analysis with comprehensive parameters.
In the previous work13, we tried illustrating gait alterations in nerve lesion model rats using comprehensive video analysis. However, in general, the potential outcomes of motion analyses are often restricted to predetermined variables provided in the analysis frameworks. For this reason, the present study further detailed how to incorporate user-defined parameters that are broadly applicable. Kinematic evaluations using video analyses may be of further use if proper parameters are implemented.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Adhesive Tape</td>
			<td>NICHIBAN CO.,LTD.</td>
			<td>SEHA25F</td>
			<td>Adhesive tape to secure thread on tails of rats for hindlimb unloading</td>
		</tr>
		<tr>
			<td>Anesthetic Apparatus for Small Animals</td>
			<td>SHINANO MFG CO.,LTD.</td>
			<td>SN-487-0T</td>
			<td></td>
		</tr>
		<tr>
			<td>Auto clicker</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>free software available to download to PC (https://www.google.com/search?client=firefox-b-1-d&#38;q=auto+clicker)</td>
		</tr>
		<tr>
			<td>CCD Camera</td>
			<td>Teledyne FLIR LLC</td>
			<td>GRAS-03K2C-C</td>
			<td>CCD (Charge-Coupled Device) cameras for video capture</td>
		</tr>
		<tr>
			<td>Cotton Thread</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>Thread to hang tails of rats from the ceiling of cage</td>
		</tr>
		<tr>
			<td>ISOFLURANE Inhalation Solution</td>
			<td>Pfizer Japan Inc.</td>
			<td>(01)14987114133400</td>
			<td></td>
		</tr>
		<tr>
			<td>Joint marker</td>
			<td>TOKYO MARUI Co., Ltd</td>
			<td>0.12g BB</td>
			<td>6 mm airsoft pellets that were used as semispherical markers with modification</td>
		</tr>
		<tr>
			<td>Kine Analyzer</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software for analysis</td>
		</tr>
		<tr>
			<td>Konishi Aron Alpha</td>
			<td>TOAGOSEI CO.,LTD.</td>
			<td>#31204</td>
			<td>Super glue to attach spherical markers on randmarks of rats</td>
		</tr>
		<tr>
			<td>Motion Recorder</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software for video recording</td>
		</tr>
		<tr>
			<td>Paint Marker</td>
			<td>MITSUBISHI PENCIL CO., LTD</td>
			<td>PX-21.13</td>
			<td>Oil based paint marker to mark toes of animals</td>
		</tr>
		<tr>
			<td>Three-dimensional motion capture apparatus (KinemaTracer for small animals)</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>3D motion analysis system that consists of four cameras (https://www.kicnet.co.jp/solutions/biosignal/animals/kinematracer-for-animal/ or https://micekc.com/en/)</td>
		</tr>
		<tr>
			<td>Three-dimensional(3D) Calculator</td>
			<td>KISSEI COMTEC CO.,LTD.</td>
			<td>N.A.</td>
			<td>Software fo marker tracking</td>
		</tr>
		<tr>
			<td>Treadmill</td>
			<td>MUROMACHI KIKAI CO.,LTD</td>
			<td>MK-685</td>
			<td>Treadmill equipped with transparent housing, electrical shocker, and speed control unit</td>
		</tr>
		<tr>
			<td>Wistar Rats (male, 7-week old)</td>
			<td>N.A.</td>
			<td>N.A.</td>
			<td>Commercially available at experimental animal sources</td>
		</tr>
	</tbody>
</table>

comprehensive understanding of inactivity-induced gait alteration in rodents

It is well known that disuse affects neural systems and that joint motions become altered; however, which outcomes properly exhibit these characteristics is still unclear. The present study describes a motion analysis approach that utilizes three-dimensional (3D) reconstruction from video captures. Using this technology, disuse-evoked alterations of walking performances were observed in rodents exposed to a simulated microgravity environment by unloading their hindlimb by their tail. After 2 weeks of unloading, the rats walked on a treadmill, and their gait motions were captured with four charge-coupled device (CCD) cameras. 3D motion profiles were reconstructed and compared to those of control subjects using the image processing software. The reconstructed outcome measures successfully portrayed distinct aspects of distorted gait motion: hyperextension of the knee and ankle joints and higher position of the hip joints during the stance phase. Motion analysis is useful for several reasons. First, it enables quantitative behavioral evaluations instead of subjective observations (e.g., pass/fail in certain tasks). Second, multiple parameters can be extracted to fit specific needs once the fundamental datasets are obtained. Despite hurdles for broader application, the disadvantages of this method, including labor intensity and cost, may be alleviated by determining comprehensive measurements and experimental procedures.

12 animals were randomly assigned to one of two groups: the unloading group (UL, n = 6) or the control group (Ctrl, n = 6). For the UL group, the hindlimbs of the animals were unloaded by the tail for 2 weeks (UL period), whereas the Ctrl group animals were left free. 2 weeks after unloading, the UL group showed a distinct gait pattern compared with the Ctrl group. Figure 1 shows normalized joint trajectories of representative subjects. During the stance phase, the UL group exhibited further...

Watch this Scientific Journal Video about Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents at JoVE.com

Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents

The present protocol describes three-dimensional motion tracking/evaluation to depict gait motion alteration of rats after exposure to a simulated disuse environment.

It is well known that disuse affects neural systems and that joint motions become altered; however, which outcomes properly exhibit these characteristics ...

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