Use of Fluorochrome-Labeled Inhibitors of Caspases to Detect Neuronal Apoptosis in the Whole-Mounted Lamprey Brain after Spinal Cord Injury

Enzyme Research, 2012 · DOI: 10.1155/2012/835731 · Published: June 16, 2012

Simple Explanation

This study introduces a new method using ﬂuorochrome-labeled inhibitors of caspases (FLICA) to detect apoptotic neuronal death in whole-mounted brain preparations. The method was tested on larval sea lampreys after spinal cord injury, a model known to induce retrograde neuronal death. FLICA reagents bind covalently to active caspases inside apoptotic cells, causing them to fluoresce and allowing for their identification.

Study Duration

2 weeks

Participants

25 larval sea lampreys

Evidence Level

Not specified

Key Findings

1
FLICA reagents can effectively detect specific activated caspases in identified neurons within the whole-mounted lamprey brain.
2
Axotomy-induced neuronal apoptosis in lampreys may involve activation of the extrinsic apoptotic pathway, as indicated by the presence of activated caspase-8.
3
Specific labeling occurred only in identifiable spinal-projecting neurons of the brainstem previously shown to undergo apoptotic neuronal death after SC transection.

Research Summary

This study introduces a new method for detecting activated caspases in whole-mounted brain preparations using FLICA reagents, demonstrating its effectiveness in identifying apoptotic neurons in the lamprey brain after spinal cord injury. The method successfully detected activated caspases in specific reticulospinal neurons known to undergo apoptosis after axotomy, supporting the hypothesis that axotomy induces apoptotic cell death in these neurons. The study also suggests that the extrinsic apoptotic pathway, involving caspase-8 activation, is likely to be activated in these neurons after spinal cord injury.

Practical Implications

New Method for Apoptosis Detection

The developed method offers a valuable tool for studying apoptosis in non-mammalian vertebrates and invertebrates where specific antibodies are lacking.

Understanding Apoptotic Pathways

Identifying the specific apoptotic pathway activated after axotomy can aid in developing therapies to protect neurons and promote regeneration.

Potential Therapeutic Targets

The involvement of the extrinsic apoptotic pathway suggests potential therapeutic targets for preventing neuronal death after spinal cord injury.

Study Limitations

1
The study is limited to the lamprey model.
2
Further research is needed to validate these findings in other species.
3
The precise mechanisms of caspase activation after axotomy require further investigation.

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