Home › Neuroscience › Stab Wound Injury of the Zebrafish Adult Telencephalon: A Method to Investigate Vertebrate Brain Neurogenesis and Regeneration
Neuroscience JoVE (Open Access) Citable · DOI

Stab Wound Injury of the Zebrafish Adult Telencephalon: A Method to Investigate Vertebrate Brain Neurogenesis and Regeneration

DOI: 10.3791/51753-v
What you'll learn
  • ✓ Perform sterile stab wound injury to adult zebrafish telencephalon
  • ✓ Prepare and section injured brain tissue using vibratome
  • ✓ Apply immunohistochemistry to analyze neurogenic and regenerative responses
  • ✓ Interpret cellular changes following mechanical brain injury
Protocol

To shed light on the cellular and molecular mechanisms of zebrafish adult neurogenesis and regeneration, we developed a protocol for invasive surgery causing mechanical injuries in the zebrafish adult telencephalon and subsequent monitoring of changes in the stabbed hemisphere by immunohistochemistry or in situ hybridization.

Difficulty
advanced
Total time
~4-6 hours per fish (injury through immunohistochemistry analysis; additional time for microscopy)
Model organism
Zebrafish (Danio rerio) adult
Biosafety
BSL-1

Steps

1
Generate stab wound in telencephalon

Perform invasive surgical puncture injury to the adult zebrafish telencephalon using a sterile needle or blade under anesthesia. This mechanical injury serves as the experimental model for investigating neurogenesis and regeneration.

▶ 01:32
2
Analyze effect of telencephalic injury

Monitor and assess morphological and cellular changes in the injured telencephalic hemisphere over specified timepoints using microscopy and documentation methods.

▶ 02:21
3
Trim agarose block and section brain

Embed fixed brain tissue in agarose, trim the block to appropriate dimensions, and generate serial coronal sections using a vibratome for downstream analysis.

▶ 04:36
4
Perform immunohistochemistry on vibratome sections

Apply primary and secondary antibodies to tissue sections to visualize neurogenic markers, proliferating cells, and regenerative responses in the injured telencephalon.

▶ 06:36
5
Interpret telencephalon stab wound results

Analyze stained sections via fluorescence microscopy to characterize cellular and molecular responses to injury, including neurogenesis and tissue regeneration patterns.

▶ 07:28
💬 Comments coming soon