Visualization of Estrogen Receptors in Colons of Mice with TNBS-Induced Crohn's Disease using Immunofluorescence

Damian Jacenik; Marta Zieli&#324;ska; Sylwia Michlewska; Jakub Fichna; Wanda  M. Krajewska

doi:10.3791/60813

Visualization of Estrogen Receptors in Colons of Mice with TNBS-Induced Crohn’s Disease usi...

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Biochemistry

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JoVE Journal Biochemistry

Visualization of Estrogen Receptors in Colons of Mice with TNBS-Induced Crohn’s Disease using Immunofluorescence

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08:02 min

March 12, 2020

DOI: 10.3791/60813-v

Damian Jacenik¹, Marta Zielińska², Sylwia Michlewska³, Jakub Fichna², Wanda M. Krajewska¹

¹Department of Cytobiochemistry, Faculty of Biology and Environmental Protection,University of Lodz, ²Department of Biochemistry, Faculty of Medicine,Medical University of Lodz, ³Laboratory of Microscopic Imaging and Specialized Biological Techniques, Faculty of Biology and Environmental Protection,University of Lodz

Overview

This protocol details a validated TNBS-induced murine model of Crohn's disease and describes methods for visualizing estrogen receptors in the colon using immunohistochemistry. The technique employs immunofluorescence on formalin-fixed colon sections embedded in paraffin.

Key Study Components

Area of Science

Neuroscience
Immunology
Pathology

Background

Estrogen signaling plays a significant role in colonic physiology.
Immunohistochemistry is effective for identifying estrogen receptors.
Understanding estrogen's impact can provide insights into colitis development.
The study utilizes a murine model to explore these mechanisms.

Purpose of Study

To establish a validated model for Crohn's disease.
To demonstrate the visualization of estrogen receptors in colon tissues.
To enhance understanding of estrogen's role in colonic health and disease.

Methods Used

TNBS-induced murine model for Crohn's disease.
Immunohistochemical visualization of estrogen receptors.
Immunofluorescence technique on formalin-fixed colon sections.
Preparation of colon samples for histological analysis.

Main Results

Successful establishment of a TNBS-induced model for Crohn's disease.
Effective visualization of estrogen receptors in colon tissues.
Demonstration of the immunofluorescence technique's reliability.
Insights into the role of estrogen in colonic inflammation.

Conclusions

The protocol provides a reliable method for studying estrogen receptors in the colon.
Findings may contribute to understanding the pathophysiology of Crohn's disease.
Further research could explore therapeutic implications of estrogen signaling.

Frequently Asked Questions

What is the significance of estrogen receptors in the colon?

Estrogen receptors play a crucial role in regulating colonic physiology and may influence the development of conditions like colitis.

How is the TNBS-induced model created?

The model is established by administering TNBS to induce inflammation in the murine colon, mimicking Crohn's disease.

What techniques are used for visualization?

Immunohistochemistry and immunofluorescence techniques are employed to visualize estrogen receptors in colon tissues.

Why is immunofluorescence preferred in this study?

Immunofluorescence allows for precise localization and visualization of specific proteins, such as estrogen receptors, in tissue samples.

What are the potential implications of this research?

Understanding estrogen's role in colonic health could lead to new therapeutic strategies for treating inflammatory bowel diseases.

The protocol presents a complete validated TNBS-induced murine model of Crohn's disease and methods for visualization of estrogen receptors by immunohistochemistry using immunofluorescence of formalin-fixed colon sections embedded in paraffin.

Increasing evidence suggests the impact of estrogen signaling on the colonic part of physiology, immunohistochemistry is a promising technique for identifying estrogen receptors in the colon and they evolve into colitis. We present a complete and validated protocol for immunohistochemical visualization of estrogen receptors in the colon, using immunofluorescence. Together with Dr.Sylwia Michlewska from Laboratory of Microscopic Imaging of the University of Lodz, we will demonstrate the procedure.

Place the colon in a petri dish. Cut the colon into one to two centimeter fragments. Place each fragment on a sponge in an appropriately labeled histological cassette.

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