Microdissection and Whole Mount Scanning Electron Microscopy Visualization of Mouse Choroid Plexus

Elien Van Wonterghem; Lien Van Hoecke; Griet Van Imschoot; Daan Verhaege; Marlies Burgelman; Roosmarijn E. Vandenbroucke

doi:10.3791/64733

Mikrodissektion und Whole-Mount-Rasterelektronenmikroskopie Visualisierung des Aderhautplexus der...

JoVE Journal
Neuroscience

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Neuroscience

Microdissection and Whole Mount Scanning Electron Microscopy Visualization of Mouse Choroid Plexus

Mikrodissektion und Whole-Mount-Rasterelektronenmikroskopie Visualisierung des Aderhautplexus der Maus

Full Text

5,569 Views

06:45 min

December 16, 2022

DOI: 10.3791/64733-v

Elien Van Wonterghem*^1,2, Lien Van Hoecke*^1,2, Griet Van Imschoot^1,2, Daan Verhaege^1,2, Marlies Burgelman^1,2, Roosmarijn E. Vandenbroucke^1,2

¹VIB Center for Inflammation Research,VIB, ²Department of Biomedical Molecular Biology,Ghent University

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article details a microdissection technique for isolating the choroid plexus (CP) from mouse brain ventricles, as well as utilizing scanning electron microscopy (SEM) to examine its cellular structure. Understanding the architecture and function of the CP is essential in neuroscience, particularly pertaining to its involvement in central nervous system health and disease.

Key Study Components

Area of Science

Neuroscience
Neuroanatomy
Cell Biology

Background

The choroid plexus is an important yet understudied tissue.
It plays a critical role in the health and pathology of the central nervous system.
Efficient isolation and characterization of the CP has implications for understanding neurological diseases.

Purpose of Study

To develop a reliable method for isolating the choroid plexus from mouse brain ventricles.
To examine the cellular structure of the choroid plexus through scanning electron microscopy.
To preserve the viability and integrity of the choroid plexus during isolation.

Methods Used

The study employs microdissection techniques to isolate the choroid plexus.
Mouse models are used during the isolation process, with specific focus on the fourth and lateral ventricles.
The methodology includes terminal anesthesia, surgical procedures, and fixation for SEM analysis.
Critical steps include the use of bromophenol blue staining for enhanced visualization, and meticulous handling to avoid contamination.

Main Results

The protocol effectively isolated the choroid plexus, enabling morphological analysis through SEM.
Observations revealed definitive structural characteristics of choroid plexus epithelial cells, including microvilli and vesicle-like structures.
These findings allow for subsequent investigations into disease-related alterations at the cellular level.

Conclusions

This study demonstrates a streamlined approach to isolate the choroid plexus for detailed structural analysis.
The insights gained can enhance understanding of the CP's role in neurological health and disease.
Overall, the method enables a significant advancement in studying this crucial yet often overlooked brain component.

Frequently Asked Questions

What are the advantages of the microdissection technique used?

The microdissection technique allows for the precise isolation of choroid plexus tissue while maintaining its integrity and functionality, minimizing contamination from surrounding tissues.

How is the choroid plexus isolated from the brain ventricles?

The isolation involves surgical dissection of the mouse brain to expose the ventricles and selectively extracting the choroid plexus using fine forceps, guided by visual cues enhanced through staining.

What types of data can be obtained from the isolated choroid plexus?

The isolated choroid plexus can be analyzed using scanning electron microscopy for morphological characteristics, as well as subjected to various biochemical assays for functional studies.

Can this method be adapted for use with other animal models?

Yes, while this protocol focuses on mouse models, it can be adapted for other species with appropriate adjustments to surgical and isolation techniques.

What are some limitations to consider when using this technique?

Care must be taken to avoid contamination and to preserve the structural integrity of the choroid plexus; any mishandling can compromise the quality of the samples and subsequent analyses.

How can the findings from this study impact research on neurological diseases?

This research provides a foundation for understanding the cellular structure and function of the choroid plexus, which may reveal critical insights into its role in the pathophysiology of neurological disorders.

Der Plexus choroideus (CP), ein in den Neurowissenschaften wenig erforschtes Gewebe, spielt eine Schlüsselrolle bei der Gesundheit und Erkrankung des zentralen Nervensystems. Dieses Protokoll beschreibt eine Mikrodissektionstechnik zur Isolierung des CP und die Verwendung von Rasterelektronenmikroskopie, um einen Gesamtüberblick über seine zelluläre Struktur zu erhalten.

Um einen besseren Einblick in die Zusammensetzung und Struktur des Plexus choroideus zu erhalten, ist eine effiziente Methode zur Isolierung des Plexus choroideus aus den Hirnventrikeln ein erster unschätzbarer Schritt. Mit dieser Mikrodissektionstechnik ist es möglich, den Plexus choroideus aus den Hirnventrikeln zu sezieren und gleichzeitig seine Funktion, Lebensfähigkeit und Struktur einzudämmen, ohne den Plexus choroideus mit dem umgebenden Gewebe zu kontaminieren. Eine wesentliche Hürde bei der Isolierung des Plexus choroideus ist die Identifizierung der Struktur, da sie noch in den Hirnventrikeln schwimmt.

Die Färbung des Plexus choroideus mit Bromphenolblau erleichtert die Isolierung. Elien Van Wonterghem, unser Laborleiter, wird das Verfahren vorführen. Beginnen Sie mit der Herstellung eines terminalen Anästhetikums, einer transkardialen Perfusionslösung und einer Lösung, die für die Rasterelektronenmikroskopie (REM) erforderlich ist, und anderen Lösungen, wie im Text erwähnt.

View the full transcript and gain access to thousands of scientific videos