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

Visualizzazione della microscopia elettronica a scansione di microdissezione e montaggio intero d...

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Neuroscience

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JoVE Journal Neuroscience

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

Visualizzazione della microscopia elettronica a scansione di microdissezione e montaggio intero del plesso coroideo del topo

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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

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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.

Il plesso coroideo (CP), un tessuto poco studiato nelle neuroscienze, svolge un ruolo chiave nella salute e nella malattia del sistema nervoso centrale. Questo protocollo descrive una tecnica di microdissezione per isolare il CP e l'uso della microscopia elettronica a scansione per ottenere una visione d'insieme della sua struttura cellulare.

Per ottenere una migliore comprensione della composizione e della struttura del plesso coroideo, un metodo efficace per isolare il plesso coroideo dai ventricoli cerebrali è un primo passo inestimabile. Con questa tecnica di microdissezione è possibile sezionare il plesso coroideo dai ventricoli cerebrali pur contenendone la funzione, la vitalità e la struttura senza contaminare il plesso coroideo con il tessuto circostante. Un ostacolo significativo nell'isolamento del plesso coroideo è identificare la struttura mentre galleggia ancora nei ventricoli cerebrali.

Colorare il plesso coroideo con blu di bromofenolo facilita l'isolamento. A dimostrare la procedura sarà Elien Van Wonterghem, il nostro responsabile di laboratorio. Inizia con la preparazione di un anestetico terminale, una soluzione di perfusione transcardica e una soluzione necessaria per la microscopia elettronica a scansione, o SEM, e altre soluzioni come menzionato nel testo.

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