Mouse genetic models to manipulate enterochromaffin cell activity - Murine Organoid ELISA

Study Purpose: The purpose of this study was to determine if the transgenic mouse lines Pet1Flp;Tac1Cre;RC::PFTox and Pet1Flp;Tac1Cre;RC::FL-hM3Dq effectively inhibit or stimulate serotonin release.

Data Collection: 5-hydroxytryptamine (5-HT) measurements released from murine intestinal organoids using ELISA (enzyme-linked immunosorbent assay).

Primary Conclusion: The conclusion of this study was that PFTox reduced the stimulated serotonin release and DCZ stimulated serotonin release in hM3Dq organoids.

Curator's Notes

Experimental Design: Murine intestinal organoids isolated from wildtype, Pet1Flp;Tac1Cre;RC::PFTox, and Pet1Flp;Tac1Cre;RC::FL-hM3Dq mice were washed twice in ice-cold PBS + 0.1% BSA. Organoid pellets were reconstituted and incubated for 15 minutes in 100 µL Ringer's (140 mM NaCl, 5 mM KCl, 2 mM CaCl2, 2 mM MgCl2, 10 mM HEPES-Na, 10 mM D-glucose, pH 7.4) + 20 μM allyl isothiocyanate (AITC) for PFTox organoids and Ringer's + 1.7 μM deschloroclozapine (DCZ) for hM3Dq organoids. After 15 minutes, supernatant was collected. Collected supernatant was diluted into half with Ringer's, and serotonin concentration was measured using a serotonin ELISA kit (Rocky Mountain Diagnotics, cat#: BA E-5900R).

Completeness: This dataset is a part of a larger study: "Mouse genetic models to manipulate enterochromaffin cell activity."

Subjects & Samples: Male (n=3) adult transgenic mice were used in this study.

Primary vs derivative data: Primary data contains a summative spreadsheet with 5-hydroxytryptamine (5-HT) measurements released from murine intestinal organoids using ELISA. Sample IDs' are displayed internally within each spreadsheet. There is no derivative data folder.