<em>In ovo</em> Expression of MicroRNA in Ventral Chick Midbrain

Carola Huber; A. Alwin Prem Anand; Manfred Mauz; Peter K&#252;nstle; Wolfgang Hupp; Bernhard Hirt; Andrea Wizenmann

doi:10.3791/50024

In ovo Expression of MicroRNA in Ventral Chick Midbrain

JoVE Journal
Neuroscience
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JoVE Journal Neuroscience

In ovo Expression of MicroRNA in Ventral Chick Midbrain

In ovo Expression of MicroRNA in Ventral Chick Midbrain

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09:19 min

September 16, 2013

DOI: 10.3791/50024-v

Carola Huber¹, A. Alwin Prem Anand¹, Manfred Mauz¹, Peter Künstle¹, Wolfgang Hupp¹, Bernhard Hirt¹, Andrea Wizenmann¹

¹Department of Experimental Embryology, Institute of Anatomy,University of Tübingen

Overview

This study presents an efficient method for selectively electroporating specific regions of the midbrain in embryos. By utilizing in ovo electroporation, researchers can better understand the role of microRNAs in brain development.

Key Study Components

Area of Science

Neuroscience
Developmental Biology

Background

Ectopic expression helps elucidate microRNA functions.
Targeting specific brain areas is challenging.
In ovo electroporation is a technique used for gene delivery.

Purpose of Study

To improve the efficiency of electroporating midbrain regions.
To investigate the role of microRNAs in brain development.

Methods Used

In ovo electroporation technique.
DNA injection into midbrain regions at specific embryonic stages.
Placement of electrodes for electrical voltage application.
Targeting of mural progenitor cells in the ventricular zone.

Main Results

Successful selective electroporation of ventral and dorsal midbrain regions.
Efficient DNA uptake by progenitor cells.
Demonstrated method enhances understanding of microRNA roles.

Conclusions

The method provides a reliable approach for studying brain development.
In ovo electroporation can be optimized for specific brain regions.

Frequently Asked Questions

What is in ovo electroporation?

In ovo electroporation is a technique used to introduce DNA into specific regions of an embryo using electrical pulses.

Why is targeting specific brain areas important?

Targeting specific areas allows researchers to study the localized effects of genes and microRNAs on brain development.

What are microRNAs?

MicroRNAs are small non-coding RNA molecules that play a crucial role in regulating gene expression.

What stages of embryonic development were used in this study?

The study focused on Hamburger Hamilton stages nine to eleven for DNA injection.

What are mural progenitor cells?

Mural progenitor cells are a type of neural stem cell located in the ventricular zone that can differentiate into various neural cell types.

Ectopic expression is one technique to elucidate the microRNAs role in brain development. However, targeting specific areas using in ovo electroporation is challenging. Here, we show an efficient way to selectively electroporate ventral and dorsal midbrain regions.

An oval electroporation begins with opening the act to expose the cheek embryo. At hamburger Hamilton, stages nine to 11 DNA is injected into the midbrain region. Then electrodes are placed left and right of the midbrain and an electrical voltage is applied.

This causes a mural progenitor cells at the ventricular zone to take up the DNA For ventral Transfection. One electrode. The anode is positioned under the membrane covering the A below the midbrain area.

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