Chick Retina Electroporation
Chick Retina Electroporation - Related Videos
06:15
In Ovo Electroporation in Embryonic Chick Retina
0 Views •
Cited by 10 •
2012
The overall goal of this video is to show how to perform targeted retinal injection and in ovo electroporation of DNA/RNA constructs into the chick embryonic retina at the Hamburger and Hamilton stage 22-23, which is about embryonic day 4 (E4). This technique is very useful to study gene expression, gene regulation, and morphological change in developing chick retina.
05:53
In vivo Electroporation of Developing Mouse Retina
0 Views •
Cited by 33 •
2011
A method for the incorporation of plasmid DNA into murine retinal cells for the purpose of performing either gain- or loss of function studies in vivo is presented. This method capitalizes on the transient increase in permeability of cell plasma membranes induced by the application of an external electrical field.
06:55
In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina
0 Views •
Cited by 42 •
2011
A method to conditionally knockdown a target protein’s expression in the adult zebrafish retina is described, which involves intravitreally injecting antisense morpholinos and electroporating them into the retina. The resulting protein is knocked down for several days, which allows testing the protein’s role in the regenerating or intact retina.
08:57
In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development
0 Views •
Cited by 5 •
2012
To assess the function and the regulation of genes during the development of midbrain dopaminergic neurons, we describe a method that involves in ovo electroporation of plasmid DNA constructs into embryonic chick ventral midbrain dopaminergic neuron progenitors. This technique can be used to achieve efficient expression of genes of interest to study different aspects of midbrain development and dopaminergic neuron differentiation.
07:38
Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation
0 Views •
Cited by 26 •
2011
This protocol describes a simple and inexpensive way to quantify the activity of cis-regulatory elements (i.e., enhancer/promoters) in living mouse retinas via explant electroporation. DNA preparation, retinal dissection, electroporation, retinal explant culture, and post-fixation analysis and quantification are described.
FAQs
The electrical pulses transiently alter retinal cell membranes, creating pores through which introduced DNA or other nucleic acids can pass. Once the membrane recovers, the transferred material can support genetic manipulation or expression during later development. This gives investigators a way to connect an experimentally changed gene state with cellular and tissue-level changes in the developing retina.
The method relies on temporary membrane permeability rather than lasting membrane disruption. Nucleic acids must enter cells during the brief access window created by the electrical pulses, after which researchers can examine developmental consequences. This mechanism makes electroporation useful for testing gene function in living embryonic tissue while preserving the continuing developmental context.
The technique supports both gene-activity manipulation and lineage-tracing experiments, depending on the nucleic acid introduced. Altering gene activity allows researchers to test whether a candidate gene influences neuronal differentiation, migration, or retinal organization. Lineage tracing instead helps reveal how labeled cells contribute to developing tissue, separating gene-function questions from cell-fate and distribution questions.
A basic experiment introduces DNA or another nucleic acid into the embryonic retina, applies brief electrical pulses, and then follows the tissue as development proceeds. Later analysis focuses on expression or developmental effects rather than only immediate uptake. This sequence links the initial genetic intervention to changes that emerge in neuronal differentiation, migration, or retinal organization.
Researchers can evaluate several levels of outcome after electroporation. They may examine whether the introduced material is expressed, how manipulated cells differentiate into neurons, where cells migrate, and whether the retina develops its expected organization. Considering these outcomes together is important because a gene may affect cell identity, positioning, or tissue architecture rather than producing a single isolated effect.
The chick embryo is useful because its developing retina can be reached directly for experimental manipulation. This accessibility lets developmental biologists alter retinal cells in an embryonic setting and observe consequences during subsequent nervous-system formation. Findings can connect gene activity with vertebrate neurobiology while providing a practical tissue context for studying retinal development.