Home › Neuroscience › Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation
Neuroscience JoVE (Open Access) Citable · DOI

Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation

DOI: 10.3791/4163-v
What you'll learn
  • ✓ Perform in utero electroporation in mouse embryos at E14.5
  • ✓ Visualize and manipulate dendrites and spines in cortex and hippocampus
  • ✓ Prepare DNA solutions and surgical instruments for electroporation
  • ✓ Execute post-surgical tissue processing for imaging analysis
Protocol

This article describes in detail a protocol to electroporate in utero the cerebral cortex and the hippocampus at E14.5 in mice. We also show that this is a valuable method to study dendrites and spines in these two cerebral regions.

Difficulty
advanced
Total time
~2-3 hours per pregnant dam (surgery + recovery); imaging analysis timeline variable
Model organism
Mouse (embryonic, E14.5)
Biosafety
BSL-1

Steps

1
Prepare DNA solution and electroporation needles

Prepare the plasmid DNA solution at appropriate concentration and load into micropipettes to create fine needles for microinjection. Ensure sterility and proper needle configuration for subsequent injection.

▶ 01:49
2
Prepare surgical field and anesthetize pregnant mouse

Anesthetize the pregnant dam, prepare the surgical site with aseptic technique, and expose the uterine horn containing embryos for access to E14.5 stage brains.

▶ 02:10
3
Inject DNA and apply electroporation pulses

Microinject DNA solution into the cerebral cortex and hippocampus of the exposed embryo, then apply controlled electrical pulses to achieve transfection of target neural cells.

▶ 03:25
4
Assess surgical outcome and process tissue

Evaluate post-operative embryo viability and uterine placement, allow dam recovery, and subsequently harvest and process brain tissue for imaging and analysis.

▶ 05:01
5
Image and analyze transfected dendrites and spines

Acquire microscopy images of electroporated neurons and quantify dendritic morphology and spine density in cortical and hippocampal regions.

▶ 05:28
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