Isolation of Intact, Whole Mouse Mammary Glands for Analysis of Extracellular Matrix Expression and Gland Morphology

Christopher Thompson; Katherine Keck; Abigail Hielscher

doi:10.3791/56512

Isolement des glandes mammaires de souris intacte, toute analyse d’Expression de la matrice extra...

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Isolation of Intact, Whole Mouse Mammary Glands for Analysis of Extracellular Matrix Expression and Gland Morphology

Isolement des glandes mammaires de souris intacte, toute analyse d’Expression de la matrice extracellulaire et la morphologie de la glande

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12:49 min

October 30, 2017

DOI: 10.3791/56512-v

Christopher Thompson¹, Katherine Keck², Abigail Hielscher²

¹Department of Biochemistry and Molecular Biology,University of Nebraska Medical Center, ²Department of Biomedical Sciences,GA-PCOM

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Overview

This article presents a protocol for isolating whole, intact mouse mammary glands to study extracellular matrix (ECM) expression and ductal morphology. The method enables researchers to analyze histological presentations of mammary tissue ex-vivo, providing insights into tumor biology.

Key Study Components

Area of Science

Neuroscience
Biology
Histology

Background

Understanding ECM expression is crucial for studying mammary gland biology.
The protocol aids in investigating ductal morphology in both healthy and diseased states.
It addresses key questions regarding stromal remodeling in tumors.
Utilizing nulliparous mice allows for controlled studies of mammary tissue.

Purpose of Study

To facilitate the analysis of mammary tissue from intact glands.
To characterize protein expression and ductal architecture.
To provide a reliable method for studying mammary gland pathology.

Methods Used

Harvesting mammary glands from 8-10 week old female nulliparous mice.
Fixation in neutral buffered formalin.
Sectioning and staining using immunohistochemistry for ECM proteins.
Pinning carcasses and using ethanol for sterilization during the procedure.

Main Results

Successful isolation of intact mammary glands for analysis.
Characterization of ECM protein expression achieved.
Insights into ductal morphology provided.
Potential applications in tumor biology highlighted.

Conclusions

The protocol is effective for studying mammary gland histology.
It allows for quantification of protein expression in various conditions.
This method can enhance understanding of mammary gland diseases.

Frequently Asked Questions

What is the main goal of this protocol?

The main goal is to facilitate the analysis of mammary tissue from intact glands ex-vivo.

How are the mammary glands prepared for analysis?

They are fixed in neutral buffered formalin and sectioned for staining.

What age and type of mice are used in this study?

8-10 week old female nulliparous mice are used for the protocol.

What techniques are employed in this study?

Immunohistochemistry is used to stain for ECM proteins.

Why is this research important?

It helps in understanding the remodeling of stromal tissue in tumors and mammary ductal architecture.

Who demonstrates this technique?

Chris Thompson, a graduate student, demonstrates the technique.

Nous présentons ici un protocole pour l’isolement des souris entière et intacte pour étudier l’expression de la matrice extracellulaire (ECM) et de la morphologie ductal des glandes mammaires. La souris #4 glandes abdominales ont été extraits des 8-10 semaines vieux nullipares les souris femelles, fixés dans du formol tamponné neutre, sectionnés et colorés par immunohistochimie pour protéines ECM.

L’objectif global de cette technique de prélèvement de glandes mammaires est de faciliter l’analyse de la présentation histologique du tissu mammaire d’une glande intacte ex-vivo. Cette méthode peut aider à répondre à des questions clés dans le domaine de la biologie tumorale, telles que la façon dont le tissu stromal est remodelé dans les tumeurs ou comment l’expression aberrante de gènes ou de protéines modifie l’architecture canalaire mammaire. Le principal avantage de cette technique est qu’elle permet aux chercheurs de caractériser et de quantifier l’expression des protéines et l’architecture canalaire dans des glandes mammaires entières de souris saines et malades.

La démonstration de cette technique sera faite par Chris Thompson, un étudiant diplômé de mon laboratoire. Après avoir sacrifié des souris nullipares femelles âgées de huit à 10 semaines selon le protocole de texte, épinglez la carcasse en position couchée et utilisez de l’éthanol à 70 % pour la saturer. Utilisez une pince pour pincer la peau juste au-dessus du pubis et entaillez-la avec de petits ciseaux chirurgicaux.

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