Immunohistochemical validation of mouse genetic strategies used to specifically activate and silence enterochromaffin cells in the intestine
Study Purpose: The purpose of this study was to determine if the transgenic mouse lines Pet1Flp;Tac1Cre;RC::PFTox, Pet1Flp;Vil1Cre;RC::PFTox, and Pet1Flp;Tac1Cre;RC::FL-hM3Dq and effectively restrict expression of the effector alleles to enterochromaffin cells.
Data Collection: Confocal imaging was performed on Nikon Ti2 microscope with Crest LFOV spinning disk and Nikon Ti microscope with Yokagawa CSU22 spinning disk. Images were assembled in ImageJ.
Primary Conclusion: The conclusion of this study was that effector allele expression is restricted to enterochromaffin cells.
Experimental Design: Mice were euthanized, the small and large intestines collected, and fixed frozen sections of 5 μm and 20μm were prepared. The microscope mounted tissue sections were stained using immunohistochemistry protocols. Antibodies against serotonin (RRID:AB_572263 and RRID:AB_1142794), GFP (RRID:AB_2307313), and mCherry (RRID:AB_10013483) were used. Confocal imaging was performed on a Nikon Ti2 microscope with Crest LFOV spinning disk and Nikon Ti microscope with Yokagawa CSU22 spinning disk. Images were assembled in ImageJ.
Completeness: This dataset is a part of the 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 is organized by the subject ID and then sample ID folders. Each folder contains raw confocal images as .nd2 files. Derivative image data (JPEG2000 and OME-TIFF) was derived from primary images (.ims). Primary images were converted with 20:1 compression to JPEG2000 (.jpx) by MBF Bioscience for web streaming and visualization on the SPARC Data Portal.