Behavioral and Histological Evaluation of Chemotherapy-Induced Gastrointestinal Disorders

Three to four months old BALB/c mice were procured from the Biogen Laboratory, Bangalore, India, and maintained in the animal house facility of Bharathidasan University. The mice were kept in an air-conditioned room at 20–22 °C with a 12-h light/dark cycle. The experimental mice were freely allowed to access standard animal feed and water. A total of 24 experimental mice were randomly divided into four groups, namely, (1) control group (N = 6), (2) cysteamine HCl group (N = 6), (3) ranitidine group (N = 6), and (4) cysteamine HCl + ranitidine group (N = 6). While mice in group 1 received normal tap water, mice in group 2 and group 4 received an intraperitoneal injection of cysteamine HCl (Sigma Aldrich, St. Louis, Missouri, USA) (60 milligrams (mg)/kilogram (Kg) body weight (BW)) for three alternate days to induce GI disorder. While a minimal dose of ranitidine can be safe and effective, in this candidate approach, the dose of ranitidine has been determined based on previous studies [36 (link)]. Thus, animals in groups 3 and 4 were orally administered ranitidine (Cadila Pharmaceuticals Limited, India) (30 mg/ Kg BW) in drinking water for 14 consecutive days. After the treatment, the experimental mice underwent behavioral experiments such as the open field test (OFT), light and dark box test (LDBT), and elevated plus maze (EPM) test. The behavioral room was equipped with a proper light setting. A video camera was placed above the center of the behavioral apparatuses. The camera was connected to a semi-automated computer system equipped with the SMART 3.0 video tracking system (Panlab, Harvard apparatus, Spain) through which digital tracking of all the animal behavioral experiments was captured (Figure 2). After the behavioral experiments, the animals were perfused, and brain tissues were processed for histology and immunostainings as earlier described by Kandasamy and colleagues [19 (link),37 (link)].
The experimental protocol involving animals was approved (Ref No. BDU/IAEC/P10/2019 dated 30.11.2019) by the Institutional Animal Ethics Committee (IAEC), Bharathidasan University under the regulation of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India.