Home › Neuroscience › A Method to Make a Craniotomy on the Ventral Skull of Neonate Rodents
Neuroscience JoVE (Open Access) Citable · DOI

A Method to Make a Craniotomy on the Ventral Skull of Neonate Rodents

DOI: 10.3791/51350-v
What you'll learn
  • ✓ Perform ventral skull craniotomy in neonatal rodents
  • ✓ Prepare anesthetized pups for acute electrophysiology and imaging
  • ✓ Conduct two-photon microscopy in exposed brainstem tissue
  • ✓ Apply electroporation and assess microvessel permeability in vivo
Protocol

A surgical method is described to expose the ventral skull in neonate rats. Using this approach it is possible to open a craniotomy to perform acute electrophysiology and two-photon microscopy experiments in the brainstem of anesthetized pups.

Difficulty
advanced
Total time
~45–60 min per pup (including anesthesia, intubation, craniotomy, and initial imaging)
Model organism
Rat (neonatal/pup)
Biosafety
BSL-1

Steps

1
Intubate anesthetized neonatal rat pup

Secure airway access in anesthetized pup to maintain respiration during surgical procedure. This prepares the animal for sustained anesthesia during craniotomy and imaging.

▶ 01:20
2
Perform ventral skull craniotomy on brainstem

Surgically open the ventral skull to expose brainstem tissue while maintaining anesthesia. This creates an optical window for acute electrophysiology and microscopy access.

▶ 03:24
3
Conduct two-photon microscopy on exposed brainstem

Acquire two-photon images through the craniotomy to visualize neural tissue and vasculature in the living pup. Positioning and imaging parameters are optimized for brainstem visualization.

▶ 04:25
4
Apply neural tracers and assess vascular permeability

Electroporate neural tracers into brainstem neurons and evaluate microvessel permeability in real time using two-photon imaging. Demonstrates functional validation of the craniotomy approach.

▶ 06:22
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