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Updated: Nov 21, 2025

Restraint to Induce Stress in Mice and Rats
Published on: December 6, 2024
Flowerpot method for rapid eye movement sleep deprivation does not induce stress as defined by elevated serum
Shafa Khan1, Manjeet Kaur1, Birendra Nath Mallick1
1School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067, India.
Abstract:
Flowerpot method of rapid eye movement sleep (REMS) deprivation (REMSD) has been most extensively used in experiments to decipher the functions of REMS. The most common but serious criticism of this method has been presumed stress experienced by the experimental animals. The lack of systematic studies with appropriate controls to resolve this issue prompted this study. We have compared serum corticosterone levels as a marker of stress in male rats under REMSD by the flowerpot method and multiple types of control conditions. Additionally, to maintain consistency and uniformity of REMSD among groups, in the same rats, we estimated brain Na-K ATPase activity, which has been consistently reported to increase upon REMSD. The most effective method was one rat in single- or multiple-platforms set-up in a pool because it significantly increased Na-K ATPase activity without elevating serum corticosterone level. More than one rat in multiple platform set-up was ineffective and must be avoided. Also, large platform- and recovery-controls must be carried out simultaneously to rule out non-specific confounding effects.

