This work was supported in part by the Ministerio de Econom&#237;a y Competitividad, Spain (DEP2012-39262 to EI-G and DEP2015-69980-P to BF-G). Thanks to Frank Mcleod Henderson Higgins from McLeod&#180;s English Centre in Asturias, Spain, for language assistance.

The methods presented in this protocol have been evaluated and approved by the animal research technical committee (reference PROAE 04/2018, Principado de Asturias, Spain).
1. Planning
<ol>
	<li>Carefully select animals for the study based on the characteristics of interest (genetically modified, pathology models, age, etc.) and apply specific adaptations to the protocol (climbing without weights, reducing the number of rungs to climb, and inclination).</li>
	<li>Identify th...

<ol>
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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volume+for+muscle+hypertrophy+and+health+outcomes:+The+most+effective+variable+in+resistance+training.">Volume for muscle hypertrophy and health outcomes: The most effective variable in resistance training.</a> Sports Medicine. 48 (3), 499-505 (2018).</li><li>Gentil, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+velocity+loss+for+monitoring+resistance+training+effort+in+a+real-world+setting.">Using velocity loss for monitoring resistance training effort in a real-world setting.</a> Applied Physiology, Nutrition, and Metabolism. 43 (8), 833-837 (2018).</li><li>Fern&#225;ndez-Sanjurjo, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Is+physical+performance+(in+mice)+increased+by+Veillonella+atypica+or+decreased+by+Lactobacillus+bulgaricus.">Is physical performance (in mice) increased by Veillonella atypica or decreased by Lactobacillus bulgaricus.</a> Journal of Sport and Health Science. 9 (3), 197-200 (2020).</li><li>Shiguemoto, G. 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V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+resistance+training+on+matrix+metalloproteinase+activity+in+skeletal+muscles+and+blood+circulation+during+aging.">Effects of resistance training on matrix metalloproteinase activity in skeletal muscles and blood circulation during aging.</a> Frontiers in Physiology. 9, 190 (2018).</li><li>Ghosh, S., Golbidi, S., Werner, I., Verchere, B. C., Laher, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selecting+exercise+regimens+and+strains+to+modify+obesity+and+diabetes+in+rodents:+an+overview.">Selecting exercise regimens and strains to modify obesity and diabetes in rodents: an overview.</a> Clinical Science. 119 (2), 57-74 (2010).</li><li>M&#244;nico-Neto, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+training+minimizes+catabolic+effects+induced+by+sleep+deprivation+in+rats.">Resistance training minimizes catabolic effects induced by sleep deprivation in rats.</a> Applied Physiology, Nutrition, and Metabolism. 40 (11), 1143-1150 (2015).</li><li>Hawley, J. A., Hargreaves, M., Joyner, M. J., Zierath, J. 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W., Krusnauskas, R., Hodson-Tole, E., Venckunas, T., Degens, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endurance+exercise+plus+overload+induces+fatigue+resistance+and+similar+hypertrophy+in+mice+irrespective+of+muscle+mass.">Endurance exercise plus overload induces fatigue resistance and similar hypertrophy in mice irrespective of muscle mass.</a> Experimental Physiology. 105 (12), 2110-2122 (2020).</li><li>Knab, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeatability+of+exercise+behaviors+in+mice.">Repeatability of exercise behaviors in mice.</a> Physiology &amp; Behavior. 98 (4), 433-440 (2009).</li><li>Konhilas, J. 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Training is an intervention with multiple applications in research, apart from the study of exercise itself. Thus, the analysis of its effect on ageing20&#160;or certain pathological conditions and physical therapy21&#160;has received much attention in recent years. In addition, numerous authors have analyzed the effect of pharmacological22&#160;or dietary21&#160;interventions on physical fitness. In this context, interest has...

Physical exercise is a determinant lifestyle factor for promoting health and decreasing the incidence of the most prevalent chronic diseases as well as some types of cancer in humans1.
Resistance exercise has raised interest because of its overwhelming relevance for health throughout life2, especially due to its benefits in counteracting age-related diseases that affect the locomotor system, such as sarcopenia, osteoporosis, etc3. Moreover, resistance exercise also affects tissues and organs not directly involved in the execution of movement, such as the brain4. This relevance in recent years has encouraged the development of resistance exercise models in animals to study the underlying tissular and molecular mechanisms, when it is not possible in humans or when the animals provide better insight and are a more controlled model.
Unlike resistance exercise in humans, for animal models&#160;researchers usually rely on forced procedures. However, aversive stimuli must be avoided in this context, mainly to preserve animal welfare, reduce stress, and decrease the severity of the experimental procedures5. It should be noted that animals enjoy exercise even in the wild6. For these reasons, it is necessary to improve adaptation to the experiment through prolonged stepwise acclimatization.
The devices, materials, and protocols used for resistance training and assessment in experimental animals must allow the precise control and modulation of numerous variables: load, volume, speed, and frequency7. They should also allow different types of muscle contractions to be performed: concentric, eccentric, or isometric. Considering the above, the protocols used should be able to specifically evaluate or train for different applications of strength: maximal strength, hypertrophy, speed, and endurance.
There are several methods of strength training, such as jumping in water8,9, weighted swimming in water10, or muscle electrostimulation11. However, static and dynamic ladders are versatile devices that are widely used12,13,14.
Resistance assessment in experimental animal models provides valuable information for many research settings, such as describing the phenotypic characteristics of genetically modified animals, evaluating the effect of different intervention protocols (dietary components supplementation, drug treatments, microbiota transplantation, etc.), or assessing the effect of training protocols. Training models provide insight into the physiology of adaptation to strength exercise, which helps to better understand the effect of exercise on health status and pathophysiology.
Consequently, there is no universal protocol for resistance training or the functional assessment of strength in animal models, so versatile protocols are needed.
The aim of this study is to identify the most relevant factors to be considered when designing and applying a protocol for resistance training and evaluation using static and dynamic ladders in animal models, as well as provide specific examples.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Dynamic ladder</td>
			<td>in-house production</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Elastic adhesive bandage 6 cm x 2.5 m</td>
			<td>BSN medical</td>
			<td>4005556</td>
			<td></td>
		</tr>
		<tr>
			<td>Gator Clip Steel NON-INSUL 10A</td>
			<td>Digikey electronics</td>
			<td>BC60ANP</td>
			<td></td>
		</tr>
		<tr>
			<td>Static ladder</td>
			<td>in-house production</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Weights</td>
			<td>in-house production</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Wire for holding weigths</td>
			<td>in-house production</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

versatility of protocols for resistance training and assessment using static and dynamic ladders in animal models

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way to gain insight into the underlying molecular mechanisms that orchestrate these adaptations. The aim of this article is to describe exercise models and training protocols designed for strength training and evaluation of resistance in animal models and provide examples. In this article, strength training and resistance evaluation are based on ladder climbing activity, using static and dynamic ladders. These devices allow a variety of training models as well as provide precise control of the main variables which determine resistance exercise: volume, load, velocity, and frequency. Furthermore, unlike resistance exercise in humans, this is a forced exercise. Thus, aversive stimuli must be avoided in this intervention to preserve animal welfare. Prior to implementation, a detailed design is necessary, along with an acclimatization and learning period. Acclimatization to training devices, such as ladders, weights, and clinical tape, as well as to the manipulations required, is necessary to avoid exercise rejection and to minimize stress. At the same time, the animals are taught to climb up the ladder, not down, to the resting area on the top of the ladder. Resistance evaluation can characterize physical strength and permit adjusting and quantifying the training load and the response to training. Furthermore, different types of strength can be evaluated. Regarding training programs, with appropriate design and device use, they can be sufficiently versatile to modulate different types of strength. Furthermore, they should be flexible enough to be modified depending on the adaptive and behavioral response of the animals or the presence of injuries. In conclusion, resistance training and assessment using ladders and weights are versatile methods in animal research.

Results with static ladder 
The progressive resistance training protocol used and described by Codina-Martinez et al.4 (Table 4) was tested in a preliminary study consisting of 7 weeks of training on a static ladder with 6-months-old wild-type C57BL6J mice (n = 4). In this preliminary study, incremental tests to assess maximal strength were performed before and after the training period. We observed a 46.4% increase in maximal strength, meaning that at the en...

Watch this Scientific Journal Video about Versatility of Protocols for Resistance Training and Assessment Using Static and Dynamic Ladders in Animal Models at JoVE.com

Versatility of Protocols for Resistance Training and Assessment Using Static and Dynamic Ladders in Animal Models

The present protocol describes resistance training and testing using static and dynamic ladders in animal models.

Resistance training is a physical exercise model with profound benefits for health throughout life. The use of resistance exercise animal models is a way ...

This resistance training protocol is significant because it can be applied in animal research to elucidate the effects of strength training on health. The main advantage of this method is that it can be applied for various strength training models and strength assessments. This technique provides an insight into physical fitness, specifically strength, and can be applied to study systemic diseases and the effect of treatments on muscle performance.It allows for training and evaluating the strength of mice, but has a mandatory requirement which is the realization of sufficient acclimatization. Visual demonstration of this method is critical because the visualization of handling, training, and assessing the resistance of the mice is more meaningful than text. Demonstrating the procedure will be Doctor Cristina Tomas-Zapico and Doctor Manuel Fernandez Sanjurjo from Doctor Benjamin Fernandez-Garcia's laboratory.To begin, carefully select animals for the study based on the characteristics of interest. Plan all issues related to training, particularly, the timetable, duration of the training period, and frequency of sessions, and draw a training table. Next, accustom the mice to stay in the resting area at the top of the ladder by leaving the animals in this area in groups of four with bedding from their cage for 15 minutes every day.Teach the mice to climb up and knock down the ladder. For this, use a static ladder and place the mouse on a rung close to the top of the ladder from where they can see the resting area and instinctively go to it. Teach them progressively to climb up from 5 rungs the first day, to 10 rungs the following day, up to 15 rungs.Next, use the same procedure with a dynamic ladder, first without movement, then with the ladder moving at 5.4 and 6.6 centimeters per second, and the mice climbing up for 2 minutes, completing 5 series. Next, prepare the following materials for the resistance exercise, weights, wire for holding weights, steel gator clip, and clinical adhesive tape. On the third day of acclimatization, cut a piece of the clinical adhesive tape and attach it around the mouse's tail to adapt it to carrying weights.On the seventh day of acclimatization, insert small weights in the wire and hook the gator clip. Clamp the gator to the clinical tape attached to the mouse's tail and place the mouse on the ladder, Immediately after the mouse has climbed the required rungs, remove the clamp and allow the mouse to rest with the clinical tape on the tail but without the weight. Before performing incremental tests to assess maximal strength, perform a warmup session consisting of three series of 10 repetitions, 10 steps per repetition without external load.Set the slope at 90 degrees for the first series, and after that at 85 degrees. Allow a rest period of 60 seconds between series. After attaching an external load of 10 grams on the mouse's tail, as demonstrated earlier, start the test with the slope set at 85 degrees and perform one series of 10 steps.Remove the weight and allow a rest period of 120 seconds in the resting area. Then perform a successive series of 10 steps increasing the external load by five grams until exhaustion. Allow the 120 second resting period between series.If a mouse fails to climb 10 steps with a particular weight load, allow for another attempt with the same load after 120 seconds of rest. If it fails again, record the weight load of the last completed series as the maximal weight load. If it succeeds in climbing with the load, continue the test with the next load.To perform an incremental test with the dynamic ladder for assessing maximal strength, clip the appropriate weight on the clinical tape and start the test at 4.2 centimeters per second. Increase the speed by 1.2 centimeters per second every 60 seconds until exhaustion and record the time as result. To perform the maximal endurance resistance test with the static ladder after a warmup session with no weights, as demonstrated previously, clip the appropriate weight on the clinical tape placed around the tail of the mouse.Then, keeping the ladder at an 85 degree slope, perform consecutive series of 10 steps until exhaustion with no resting time between each series and record the number of rungs climbed. To perform the maximal endurance resistance test with the dynamic ladder, set the slope at 85 degrees and speed at 4.2 centimeters per second. For warming up, perform three series of 100 steps without external load and allow rest period of 60 seconds between series.After the warmup, allow a 120 second rest period before initiating the test. Then clip the gator with the external load, place the mouse on the running ladder, and start the stopwatch. The test ends when the mouse is no longer able to maintain the rate of ascent.For resistance training with the static ladder, start the training session in the resting area and perform the warmup session without external load. Then clip the gator with the weight on the clinical tape and gently place the mouse 10 to 20 rungs below the resting place. Allow the mouse to grip the rung and climb to the resting area.Repeat this process until the number of rungs in the series is completed. Remove the weight from the mouse tail and wait for 120 seconds until the next series. Increase the number of steps and the maximum weight load of the series throughout the training period following the training plan.For resistance training with the dynamic ladder, after setting the slope to 85 degrees, close the door to the resting area and start the ladder at the desired speed. After a warmup procedure, when the mouse is in the resting area, clip the gator with the weight on the clinical tape. Alternatively, the weight can be attached when the mouse is on the ladder.Then gently place the mouse at the top of the moving ladder with the weight on the tail and allow the mice to grip the rung and climb. When the number of rungs in this series is reached, the weights are removed and the door is opened so that the animal can go to the resting area. The rest time is 120 seconds before the next series.Repeat this procedure until the training session is completed. The effect on resistance after six weeks of strength training on a dynamic ladder is shown here. The variation in maximal strength shows a significant increase after training in the strength and endurance resistance groups, while this parameter did not change in the control group.Endurance resistance measured at the end of the training period was significantly higher in the endurance resistance group as compared to that in the strength and control groups. Proper acclimatization, including learning to climb and adapting to the clinical tape and weights is important. This procedure allows researchers to design various types of resistance training interventions.Strength is relevant to human health and aging. Researchers can use these methods to explore the effects of exercise and pharmacological interventions on resistance.

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Research

JoVE Journal

Biology

2.3K vistas.  University of Oviedo. Este protocolo de entrenamiento de resistencia es importante porque se puede aplicar en la investigación con animales para dilucidar los efectos del entrenamiento de fuerza en la salud. La principal ventaja de este método es que se puede aplicar para varios modelos de entrenamiento de fuerza y evaluaciones de fuerza. Esta técnica proporciona una visión de la aptitud física, específicamente la fuerza, y se puede aplicar para estudiar enfermedades sistémicas y el efecto de los tratamient...