A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

Summary

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

Abstract

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed as a water tank with narrow platforms on the bottom. The model rats were put into the tank and stood on the platforms for 14 h (18:00 - 8:00) per day for a consecutive 21 days, with a blank control group set for contrast. At the end of modeling, rats in the model group showed an obvious fatigued appearance. To assess the model, we performed several behavioral tests, including the open field test (OFT), the elevated plus maze (EPM) test, and the exhaustive swimming (ES) test. The results showed that anxiety, spatial cognition impairment, poor muscle performance, and declined voluntary activity presented in model rats confirm the diagnosis of central fatigue. Changes of the central neurotransmitters also verified the result. In conclusion, the model successfully simulated central fatigue, and future study with the model may help reveal the pathological mechanism of the disease.

Introduction

Fatigue is one of the main factors that threaten human health¹. In the past decades, various researches have proved that fatigue is peripherally-triggered but centrally-driven, and always accompanied with emotional and cognitive disorders. Italian physiologist A. Mosso first proposed the word Central Fatigue². It is generally defined as limited voluntary activity and cognition impairment due to dysfunction of impulse transmission in the central nervous system (CNS)³. Compared with peripheral muscle fatigue, central fatigue emphasizes changes in the CNS, as well as the consequent emotional/behavior....

Protocol

All the animals were maintained in accordance with the guidelines by the Chinese legislation on the ethical use and care of laboratory animals.

1. Pre-modeling Preparation

1. Laboratory preparation
   1. Run the UV lamp for at least 30 min before the experiment.
   2. Control the lab temperature at 25 ± 3 °C, and relative humidity around 30%.
   3. Turn on the lab light at 6:00 and turn it off at 18:00 to establish a 12 h/12 h light/dark cycle.
2. Multiple Platform box construction
   1. Construct an opaque plastic tank without a cover of 110 ....

Results

We describe a rat model of central fatigue using MMPM. 24 Wistar rats are randomly divided into the control group and the model group, with 12 rats in each group. The model apparatus is designed as a water tank with narrow platforms on the bottom (Figure 1). Model Rats stand on the platforms for 14 h per day, including partial sleep time, for 21 days (Figure 2).

Behavio.......

Discussion

The MMPM is originally designed for sleep deprivation⁹. Rats are launched into a water tank with platforms fixed on the bottom. Driven by the instinctive fear of water, rats remain standing on the platforms and no sleep occurs. The study shows that different hours of sleep deprivation lead to various changes in rat behavior and mood, including recognition impairment¹⁰, negative emotions¹¹, and central fatigue. Some researchers prove that chronic slee.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
multiple platform sleep deprivation water tank	Customization,it is provided by the neuroimmunological laboratory of Beijing University of Chinese Medicine		110cm x 60cm x 40cm. There are 15 plastic small platforms at the bottom. The small platform is 6.5cm in diameter and 8cm high
Wistar rats	Beijing Weitong Lihua Experimental Animal Technology Company	license number SYXK (Beijing) 2016-0011	Use 32 Wistar healthy male rats ,8 week old (200-210 g)
Agilent 1100LC high performance liquid chromatograph	Agilent	G1379A, G1311A, G1313A , G1316A	G1379A, G1311A type chromatographic pump, G1313A automatic sampler, G1316A column temperature box
DECADE II SDC electrochemical detector	Dutch ANTEC company		glassy carbon electrode, Ag/AgCl reference electrode, workstations (Clarity CHS)
Biofuge Stratos high-speed refrigeration centrifuge	HERAEUS
VCX130 ultrasonic fracturing instrument	SONICS
ACS-ZEAS electronic scale	Phos technology development, Beijing.		The weight of the weighing rats can be accurate to 0.1g.
Open Field Box	Customization,it is provided by the neuroimmunological laboratory of Beijing University of Chinese Medicine		wooden box of open field  100 cm by 100 cm x 40 cm, inside wall and bottom as the gray.The bottom is divided into 25 equal area squares, each of which is 20cm x 20cm, and the 16 grids along the outer wall are the external ones, and the other 9 grids are central.The camera is mounted above the median.
Elevated Plus-maze	Beijing zhongshi dechuang technology development co. LTD.		The open arms and close  arms of the cross are composed of 30cm x 5cm x 15cm, and the central area is 5cm x 5cm, with a camera mounted above the center and 45cm high.
rat swimming bucket.	Zhenhua biological instrument equipment co., LTD. Anhui,China.		The volume of plastic drum is 70cm x 30cm x 110cm, which is used for swimming in rats.
Thermometer	Shiya instrument co., LTD., changzhou,China.		Control water temperature
Small water pump	Xincheng technology co., LTD., chengdu,China.		Used for water tank and swimming behavior.
Ethovition3.0 behavioral software.	Nuldus,Netherlands		Measurement analysis of rat behavior videos.