Supplementary data and analysis code for: Enteric serotonin and the 5-HT3 gut-to-brain axis in chronic environmental heat stress — a loss-of-function and gain-of-function study in rats

Supplementary data and analysis code for the manuscript "Enteric serotonin and the 5-HT3 gut-to-brain axis in chronic environmental heat stress: a loss-of-function and gain-of-function study in rats" by Mohamed E. Elbeeh (Department of Biology, Jamoum University College, Umm Al-Qura University, 21955 Makkah, Saudi Arabia).

STUDY. Wistar rats underwent a 14-day chronic environmental heat-stress protocol under thermoneutral or heat conditions. The loss-of-function arm received the 5-HT3 receptor antagonist ondansetron (1 mg/kg/day orally); the gain-of-function arm received intracolonic serotonin (0.5 mL of a 1e-5 M solution per day) under thermoneutral conditions. Vehicle, drug-only, glutamine positive-control arms and a confirmatory female cohort were included (101 animals in total). The primary endpoint was in-vivo intestinal permeability measured as plasma 4-kDa FITC-dextran. Secondary endpoints covered barrier and translocation markers, colonic and hippocampal cytokines, redox and oxidative-stress measures, regional monoamines by HPLC, qPCR gene expression, ex-vivo colonic contractility, and a behavioural battery.

CONTENTS.

DataTemplate_heat_5HT3_gutbrain_FILLED.xlsx: the complete structured source workbook (animal log, exposure and welfare records, every endpoint sheet, and the reagent, ELISA-kit and qPCR-primer metadata sheets).

analysis_heat_5HT3_gutbrain.py: the analysis script that generates every value reported in the manuscript.

stats_full.json: the full machine-readable statistical output (group summaries, effect sizes with bootstrap confidence intervals, family-wise Sidak and Holm corrected p-values, and Benjamini-Hochberg FDR values within endpoint families).

analysis_log.txt: the execution log of the analysis run.

Supplementary_S8_Individual_animal_data.xlsx: individual animal-level values for all endpoints (101 animals, 68 endpoints).

REPRODUCIBILITY. Running the script on the workbook regenerates the complete statistical output; no value in the manuscript was entered by hand. The study was not prospectively registered; the full analysis plan is described in the Methods section of the manuscript.

ETHICS. Approved by the Biomedical Research Ethics Committee, Umm Al-Qura University (approval no. HAPO-02-K-012-2023-03-1709, March 2023). Reported in accordance with ARRIVE 2.0.

ACCESS. The files in this record are restricted. Access is granted on reasonable request to the depositor for non-commercial academic use, consistent with the data-availability statement of the manuscript.